Search results for: pathogen propagation

Authors: Celine Shi Ying Lee, Jong Jian Lee

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Background: Infectious keratitis is one of the leading causes of blindness worldwide. Prompt treatment with effective medication will control the infection early, preventing corneal scarring and visual loss. fluoroquinolones ophthalmic medication is used because of its broad-spectrum properties, potency, good intraocular penetration, and low toxicity. The study aims to evaluate the treatment outcome of corneal ulcers using Levofloxacin 1.5% ophthalmic solution (LVFX) with adjuvant oral ciprofloxacin when indicated and apply this treatment strategy in primary health care as first-line treatment. Methods: Patients with infective corneal ulcer treated in an eye center were recruited. Inclusion criteria includes Corneal infection consistent with bacterial keratitis, single or multiple small corneal ulcers. Treatment regime: LVFX hourly for the first 2 days, 2 hourly from the 3rd day, and 3 hourly on the 5th day of review. Adjuvant oral ciprofloxacin 500mg BD was administered for 5 days if there were multiple corneal ulcers or when the location of the cornea ulcer was central or paracentral. Results: 47 subjects were recruited. There were 16 (34%) males and 31 (66%) females. 40 subjects (85%) were contact lens (CL) related to corneal ulcer, and 7 subjects (15%) were non-contact lens related. 42 subjects (89%) presented with one ulcer, of which 20 of them (48%) needed adjuvant therapy. 5 subjects presented with 2 or 3 ulcers, of which 3 needed adjuvant therapy. A total of 23 subjects (49%) was given adjuvant therapy (oral ciprofloxacin 500mg BD for 5 days).21 of them (91%) were CL related. All subjects recovered fully, and the average duration of treatment was 3.7 days, with 49% of the subjects resolved on the 3rd day, 38% on the 5thday of and 13% on the 7thday. All subjects showed symptoms of relief of pain, light-sensitivity, and redness on the 3rd day with full visual recovery post-treatment. No adverse drug reactions were recorded. Conclusion: Our treatment regime demonstrated good clinical outcome as first-line treatment for corneal ulcers. A corneal ulcer is a common eye condition in Singapore, mainly due to CL wear. Pseudomonas aeruginosa is the most frequent and potentially sight-threatening pathogen involved in CL related corneal ulcer. Coagulase-negative Staphylococci, Staphylococcus aureus, and Streptococcus Pneumoniae were seen in non-CL users. All these bacteria exhibit good sensitivity rates to ciprofloxacin and levofloxacin. It is therefore logical in our study to use LVFX Eyedrops and adjuvant ciprofloxacin oral antibiotics when indicated as first line treatment for most corneal ulcers. Our study of patients, both CL related and non-CL related, have shown good clinical response and full recovery using the above treatment strategy. There was also a full restoration of visual acuity in all the patients. Eye-trained primary Healthcare practitioners can consider adopting this treatment strategy as first line treatment in patients with corneal ulcers. This is relevant during the COVID pandemic, where hospitals are overwhelmed with patients and in regions with limited access to specialist eye care. This strategy would enable early treatment with better clinical outcome.

Keywords: corneal ulcer, levofloxacin hydrate, treatment strategy, ciprofloxacin

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

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

Keywords: antifungals, antioxidant system, benzo derivatives, chemosensitization

Procedia PDF Downloads 240

Authors: Shahin A. Abdel-Naby, James P. Colgan, Michael S. Pindzola

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A time-dependent close-coupling method is developed to calculate a total, double and triple autoionization rates for hollow atomic ions of four-electron systems. This work was motivated by recent observations of the four-electron Auger process in near K-edge photoionization of C+ ions. The time-dependent close-coupled equations are solved using lattice techniques to obtain a discrete representation of radial wave functions and all operators on a four-dimensional grid with uniform spacing. Initial excited states are obtained by relaxation of the Schrodinger equation in imaginary time using a Schmidt orthogonalization method involving interior subshells. The radial wave function grids are partitioned over the cores on a massively parallel computer, which is essential due to the large memory requirements needed to store the coupled-wave functions and the long run times needed to reach the convergence of the ionization process. Total, double, and triple autoionization rates are obtained by the propagation of the time-dependent close-coupled equations in real-time using integration over bound and continuum single-particle states. These states are generated by matrix diagonalization of one-electron Hamiltonians. The total autoionization rates for each L excited state is found to be slightly above the single autoionization rate for the excited configuration using configuration-average distorted-wave theory. As expected, we find the double and triple autoionization rates to be much smaller than the total autoionization rates. Future work can be extended to study electron-impact triple ionization of atoms or ions. The work was supported in part by grants from the American University of Sharjah and the US Department of Energy. Computational work was carried out at the National Energy Research Scientific Computing Center (NERSC) in Berkeley, California, USA.

Keywords: hollow atoms, autoionization, auger rates, time-dependent close-coupling method

Procedia PDF Downloads 138

Authors: Abirami Baleswaran, Christel Couderc, Loubnah Belahcen, Jean Dayde, Hélène Tormo, Gwénaëlle Jard

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Cheeses are often reported to be spoiled by Pseudomonas spp., responsible for defects in appearance, texture, taste, and smell, leading to their non-marketing and even their destruction. Despite preventive actions, problems linked to Pseudomonas spp. are difficult to control by the lack of knowledge and control of these contaminants during the cheese manufacturing. Lactic goat cheese producers are not spared by this problem and are looking for solutions to decrease the number of spoiled cheeses. To explore different hypotheses, experiments are needed. However, cheese-making experiments at the pilot scale are expensive and time consuming. Thus, there is a real need to develop a miniature cheeses model system under controlled conditions. In a previous study, several miniature cheese models corresponding to different type of commercial cheeses have been developed for different purposes. The models were, for example, used to study the influence of milk, starters cultures, pathogen inhibiting additives, enzymatic reactions, microflora, freezing process on cheese. Nevertheless, no miniature model was described on the lactic goat cheese. The aim of this work was to develop a miniature cheese model system under controlled laboratory conditions which resembles commercial lactic goat cheese to study Pseudomonas spp. spoilage during the manufacturing and ripening process. First, a protocol for the preparation of miniature cheeses (3.5 times smaller than a commercial one) was designed based on the cheese factorymanufacturing process. The process was adapted from “Rocamadour” technology and involves maturation of pasteurized milk, coagulation, removal of whey by centrifugation, moulding, and ripening in a little scale cellar. Microbiological (total bacterial count, yeast, molds) and physicochemical (pH, saltinmoisture, moisture in fat-free)analyses were performed on four key stages of the process (before salting, after salting, 1st day of ripening, and end of ripening). Factory and miniature cheeses volatilomewere also obtained after full scan Sift-MS cheese analysis. Then, Pseudomonas spp. strains isolated from contaminated cheeses were selected on their origin, their ability to produce pigments, and their enzymatic activities (proteolytic, lecithinasic, and lipolytic). Factory and miniature curds were inoculated by spotting selected strains on the cheese surface. The expression of cheese spoilage was evaluated by counting the level of Pseudomonas spp. during the ripening and by visual observation and under UVlamp. The physicochemical and microbiological compositions of miniature cheeses permitted to assess that miniature process resembles factory process. As expected, differences involatilomes were observed, probably due to the fact that miniature cheeses are made usingpasteurized milk to better control the microbiological conditions and also because the little format of cheese induced probably a difference during the ripening even if the humidity and temperature in the cellar were quite similar. The spoilage expression of Pseudomonas spp. was observed in miniature and factory cheeses. It confirms that the proposed model is suitable for the preparation of miniature cheese specimens in the spoilage study of Pseudomonas spp. in lactic cheeses. This kind of model could be deployed for other applications and other type of cheese.

Keywords: cheese, miniature, model, pseudomonas spp, spoilage

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Authors: Jone Ibarruri, Mikel Manso, Marta Cebrián

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A high amount of food is lost or discarded in the World every year. In addition, in the last decades, an increasing demand of new alternative and sustainable sources of proteins and other valuable compounds is being observed in the food and feeding sectors and, therefore, the use of food by-products as nutrients for these purposes sounds very interesting from the environmental and economical point of view. However, the direct use of discarded fruit and vegetables that present, in general, a low protein content is not interesting as feeding ingredient except if they are used as a source of fiber for ruminants. Especially in the case of aquaculture, several alternatives to the use of fish meal and other vegetable protein sources have been extensively explored due to the scarcity of fish stocks and the unsustainability of fishing for these purposes. Fish mortality is also of great concern in this sector as this problem highly reduces their economic feasibility. So, the development of new functional and natural ingredients that could reduce the need for vaccination is also of great interest. In this work, several fermentation tests were developed at lab scale using a selected mixture of fruit and vegetable discards from a wholesale market located in the Basque Country to increase their protein content and also to produce some bioactive extracts that could be used as additives in aquaculture. Fruit and vegetable mixtures (60/40 ww) were centrifugated for humidity reduction and crushed to 2-5 mm particle size. Samples were inoculated with a selected Rhizopus oryzae strain and fermented for 7 days in controlled conditions (humidity between 65 and 75% and 28ºC) in Petri plates (120 mm) by triplicate. Obtained results indicated that the final fermented product presented a twofold protein content (from 13 to 28% d.w). Fermented product was further processed to determine their possible functionality as a feed additive. Extraction tests were carried out to obtain an ethanolic extract (60:40 ethanol: water, v.v) and remaining biomass that also could present applications in food or feed sectors. The extract presented a polyphenol content of about 27 mg GAE/gr d.w with antioxidant activity of 8.4 mg TEAC/g d.w. Remining biomass is mainly composed of fiber (51%), protein (24%) and fat (10%). Extracts also presented antibacterial activity according to the results obtained in Agar Diffusion and to the Minimum Inhibitory Concentration (MIC) tests determined against several food and fish pathogen strains. In vitro, digestibility was also assessed to obtain preliminary information about the expected effect of extraction procedure on fermented product digestibility. First results indicated that remaining biomass after extraction doesn´t seem to improve digestibility in comparison to the initial fermented product. These preliminary results show that fermented fruit and vegetables can be a useful source of functional ingredients for aquaculture applications and a substitute of other protein sources in the feeding sector. Further validation will be also carried out through “in vivo” tests with trout and bass.

Keywords: fungal solid state fermentation, protein increase, functional extracts, feed ingredients

Procedia PDF Downloads 51

Authors: Jovana Grahovac, Ivana Pajcin, Natasa Lukic, Jelena Dodic, Aleksandar Jokic

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To obtain purified biomass to be used in the plant pathogen biocontrol or as soil biofertilizer, it is necessary to eliminate residual broth components at the end of the fermentation process. The main drawback of membrane separation techniques is permeate flux decline due to the membrane fouling. Fouling mitigation measures increase the pressure drop along membrane channel due to the increased resistance to flow of the feed suspension, thus increasing the hydraulic power drop. At the same time, these measures lead to an increase in the permeate flux due to the reduced resistance of the filtration cake on the membrane surface. Because of these opposing effects, the energy efficiency of fouling mitigation measures is limited, and the justification of its application is provided by information on a reducing specific energy consumption compared to a case without any measures employed. In this study, the influence of static mixer (Kenics) and air-sparging (two-phase flow) on reduction of specific energy consumption (ER) was investigated. Cultivation Bacillus velezensis was carried out in the 3-L bioreactor (Biostat® Aplus) containing 2 L working volume with two parallel Rushton turbines and without internal baffles. Cultivation was carried out at 28 °C on at 150 rpm with an aeration rate of 0.75 vvm during 96 h. The experiments were carried out in a conventional cross-flow microfiltration unit. During experiments, permeate and retentate were recycled back to the broth vessel to simulate continuous process. The single channel ceramic membrane (TAMI Deutschland) used had a nominal pore size 200 nm with the length of 250 mm and an inner/external diameter of 6/10 mm. The useful membrane channel surface was 4.33×10⁻³ m². Air sparging was brought by the pressurized air connected by a three-way valve to the feed tube by a simple T-connector without diffusor. The different approaches to flux improvement are compared in terms of energy consumption. Reduction of specific energy consumption compared to microfiltration without fouling mitigation is around 49% and 63%, for use of two-phase flow and a static mixer, respectively. In the case of a combination of these two fouling mitigation methods, ER is 60%, i.e., slightly lower compared to the use of turbulence promoter alone. The reason for this result can be found in the fact that flux increase is more affected by the presence of a Kenics static mixer while sparging results in an increase of energy used during microfiltration. By comparing combined method with turbulence promoter flux enhancement method ER is negative (-7%) which can be explained by increased power consumption for air flow with moderate contribution to the flux increase. Another confirmation for this fact can be found by comparing energy consumption values for combined method with energy consumption in the case of two-phase flow. In this instance energy reduction (ER) is 22% that demonstrates that turbulence promoter is more efficient compared to two phase flow. Antimicrobial activity of Bacillus velezensis biomass against phytopathogenic isolates Xanthomonas campestris was preserved under different fouling reduction methods.

Keywords: Bacillus velezensis, microfiltration, static mixer, two-phase flow

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Authors: Addin Osman, Anwar Ali Yahya, Mohammed Basit Kamal

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Datasets or collections are becoming important assets by themselves and now they can be accepted as a primary intellectual output of a research. The quality and usage of the datasets depend mainly on the context under which they have been collected, processed, analyzed, validated, and interpreted. This paper aims to present a collection of program educational objectives mapped to student’s outcomes collected from self-study reports prepared by 32 engineering programs accredited by ABET. The manual mapping (classification) of this data is a notoriously tedious, time consuming process. In addition, it requires experts in the area, which are mostly not available. It has been shown the operational settings under which the collection has been produced. The collection has been cleansed, preprocessed, some features have been selected and preliminary exploratory data analysis has been performed so as to illustrate the properties and usefulness of the collection. At the end, the collection has been benchmarked using nine of the most widely used supervised multiclass classification techniques (Binary Relevance, Label Powerset, Classifier Chains, Pruned Sets, Random k-label sets, Ensemble of Classifier Chains, Ensemble of Pruned Sets, Multi-Label k-Nearest Neighbors and Back-Propagation Multi-Label Learning). The techniques have been compared to each other using five well-known measurements (Accuracy, Hamming Loss, Micro-F, Macro-F, and Macro-F). The Ensemble of Classifier Chains and Ensemble of Pruned Sets have achieved encouraging performance compared to other experimented multi-label classification methods. The Classifier Chains method has shown the worst performance. To recap, the benchmark has achieved promising results by utilizing preliminary exploratory data analysis performed on the collection, proposing new trends for research and providing a baseline for future studies.

Keywords: ABET, accreditation, benchmark collection, machine learning, program educational objectives, student outcomes, supervised multi-class classification, text mining

Procedia PDF Downloads 150

Authors: Jinlai Bian, Zailin Yang, Guanxixi Jiang, Xinzhu Li

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The problem of elastic wave propagation in inhomogeneous medium has always been a classic problem. Due to the frequent occurrence of earthquakes, many economic losses and casualties have been caused, therefore, to prevent earthquake damage to people and reduce damage, this paper studies the dynamic response around the circular inclusion in the whole space with inhomogeneous modulus, the inhomogeneity of the medium is reflected in the shear modulus of the medium with the spatial position, and the density is constant, this method can be used to solve the problem of the underground buried pipeline. Stress concentration phenomena are common in aerospace and earthquake engineering, and the dynamic stress concentration factor (DSCF) is one of the main factors leading to material damage, one of the important applications of the theory of elastic dynamics is to determine the stress concentration in the body with discontinuities such as cracks, holes, and inclusions. At present, the methods include wave function expansion method, integral transformation method, integral equation method and so on. Based on the complex function method, the Helmholtz equation with variable coefficients is standardized by using conformal transformation method and wave function expansion method, the displacement and stress fields in the whole space with circular inclusions are solved in the complex coordinate system, the unknown coefficients are solved by using boundary conditions, by comparing with the existing results, the correctness of this method is verified, based on the superiority of the complex variable function theory to the conformal transformation, this method can be extended to study the inclusion problem of arbitrary shapes. By solving the dynamic stress concentration factor around the inclusions, the influence of the inhomogeneous parameters of the medium and the wavenumber ratio of the inclusions to the matrix on the dynamic stress concentration factor is analyzed. The research results can provide some reference value for the evaluation of nondestructive testing (NDT), oil exploration, seismic monitoring, and soil-structure interaction.

Keywords: circular inclusions, complex variable function, dynamic stress concentration factor (DSCF), inhomogeneous medium

Procedia PDF Downloads 123

Authors: Jae-Yong Park, Yeo-Kyeong Lee, Hee-Sun Kim

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Purpose: This study aims at investigating the mechanical behaviors of mastication, according to various teeth movement. There are three masticatory cases which are general case and 2 cases of teeth movement. General case includes the common arrange of all teeth and 2 cases of teeth movement are that one is the half movement location case of molar teeth in no. 14 tooth seat after extraction of no. 14 tooth and the other is no. 14 tooth seat location case of molar teeth after extraction in the same case before. Materials and Methods: In order to analyze these cases, 3 dimensional finite element (FE) model of the skull were generated based on computed tomography images, 964 dicom files of 38 year old male having normal occlusion status. An FE model in general occlusal case was used to develop CAE procedure. This procedure was applied to FE models in other occlusal cases. The displacement controls according to loading condition were applied effectively to simulate occlusal behaviors in all cases. From the FE analyses, von Mises stress distribution of skull and teeth was observed. The von Mises stress, effective stress, had been widely used to determine the absolute stress value, regardless of stress direction and yield characteristics of materials. Results: High stress was distributed over the periodontal area of mandible under molar teeth when the mandible was transmitted to the coronal-apical direction in the general occlusal case. According to the stress propagation from teeth to cranium, stress distribution decreased as the distribution propagated from molar teeth to infratemporal crest of the greater wing of the sphenoid bone and lateral pterygoid plate in general case. In 2 cases of teeth movement, there were observed that high stresses were distributed over the periodontal area of mandible under teeth where they are located under the moved molar teeth in cranium. Conclusion: The predictions of the mechanical behaviors of general case and 2 cases of teeth movement during the masticatory process were investigated including qualitative validation. The displacement controls as the loading condition were applied effectively to simulate occlusal behaviors in 2 cases of teeth movement of molar teeth.

Keywords: cranium, finite element analysis, mandible, masticatory action, occlusal force

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Authors: A. Falenski, A. Kaesbohrer, M. Filter

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Introduction: Risk assessments can be performed in various ways and in different degrees of complexity. In order to assess risks associated with imported foods additional information needs to be taken into account compared to a risk assessment on regional products. The present review is an overview on currently available best practise approaches and data sources used for food import risk assessments (IRAs). Methods: A literature review has been performed. PubMed was searched for articles about food IRAs published in the years 2004 to 2014 (English and German texts only, search string “(English [la] OR German [la]) (2004:2014 [dp]) import [ti] risk”). Titles and abstracts were screened for import risks in the context of IRAs. The finally selected publications were analysed according to a predefined questionnaire extracting the following information: risk assessment guidelines followed, modelling methods used, data and software applied, existence of an analysis of uncertainty and variability. IRAs cited in these publications were also included in the analysis. Results: The PubMed search resulted in 49 publications, 17 of which contained information about import risks and risk assessments. Within these 19 cross references were identified to be of interest for the present study. These included original articles, reviews and guidelines. At least one of the guidelines of the World Organisation for Animal Health (OIE) and the Codex Alimentarius Commission were referenced in any of the IRAs, either for import of animals or for imports concerning foods, respectively. Interestingly, also a combination of both was used to assess the risk associated with the import of live animals serving as the source of food. Methods ranged from full quantitative IRAs using probabilistic models and dose-response models to qualitative IRA in which decision trees or severity tables were set up using parameter estimations based on expert opinions. Calculations were done using @Risk, R or Excel. Most heterogeneous was the type of data used, ranging from general information on imported goods (food, live animals) to pathogen prevalence in the country of origin. These data were either publicly available in databases or lists (e.g., OIE WAHID and Handystatus II, FAOSTAT, Eurostat, TRACES), accessible on a national level (e.g., herd information) or only open to a small group of people (flight passenger import data at national airport customs office). In the IRAs, an uncertainty analysis has been mentioned in some cases, but calculations have been performed only in a few cases. Conclusion: The current state-of-the-art in the assessment of risks of imported foods is characterized by a great heterogeneity in relation to general methodology and data used. Often information is gathered on a case-by-case basis and reformatted by hand in order to perform the IRA. This analysis therefore illustrates the need for a flexible, modular framework supporting the connection of existing data sources with data analysis and modelling tools. Such an infrastructure could pave the way to IRA workflows applicable ad-hoc, e.g. in case of a crisis situation.

Keywords: import risk assessment, review, tools, food import

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Authors: K. S. Al-Mugren, A. Yahya, S. Alodah, R. Alharbi, S. H. Almsaid , A. Alqahtani, H. Jaber, A. Basaqer, N. Alajra, N. Almoghati, A. Alsalman, Khalid Alharbi

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Context: The production of biofertilizers from naturally occurring microorganisms is an area of research that aims to enhance agricultural practices by utilizing local resources. This research project focuses on isolating and screening indigenous microorganisms with PK-fixing and phosphate solubilizing characteristics from local sources. Research Aim: The aim of this project is to develop a biofertilizer product using indigenous microorganisms and composted agro waste as a carrier. The objective is to enhance crop productivity and soil fertility through the application of biofertilizers. Methodology: The research methodology includes several key steps. Firstly, indigenous microorganisms will be isolated from local resources using the ten-fold serial dilutions technique. Screening assays will be conducted to identify microorganisms with phosphate solubilizing and PK-fixing activities. Agro-waste materials will be collected from local agricultural sources, and composting experiments will be conducted to convert them into organic matter-rich compost. Physicochemical analysis will be performed to assess the composition of the composted agro-waste. Gamma and X-ray irradiation will be used to sterilize the carrier material. The sterilized carrier will be tested for sterility using the ten-fold serial dilutions technique. Finally, selected indigenous microorganisms will be developed into biofertilizer products. Findings: The research aims to find suitable indigenous microorganisms with phosphate solubilizing and PK-fixing characteristics for biofertilizer production. Additionally, the research aims to assess the suitability of composted agro waste as a carrier for biofertilizers. The impact of gamma irradiation sterilization on pathogen elimination will also be investigated. Theoretical Importance: This research contributes to the understanding of utilizing indigenous microorganisms and composted agro waste for biofertilizer production. It expands knowledge on the potential benefits of biofertilizers in enhancing crop productivity and soil fertility. Data Collection and Analysis Procedures: The data collection process involves isolating indigenous microorganisms, conducting screening assays, collecting and composting agro waste, analyzing the physicochemical composition of composted agro waste, and testing carrier sterilization. The analysis procedures include assessing the abilities of indigenous microorganisms, evaluating the composition of composted agro waste, and determining the sterility of the carrier material. Conclusion: The research project aims to develop biofertilizer products using indigenous microorganisms and composted agro waste as a carrier. Through the isolation and screening of indigenous microorganisms, the project aims to enhance crop productivity and soil fertility by utilizing local resources. The research findings will contribute to the understanding of the suitability of composted agro waste as a carrier and the efficacy of gamma irradiation sterilization. The research outcomes will have theoretical importance in the field of biofertilizer production and agricultural practices.

Keywords: biofertilizer, microorganisms, agro waste, nuclear technologies

Procedia PDF Downloads 94

Authors: Anja I. Petrov, Milica B. Veljković, Marija M. Ćorović, Ana D. Milivojević, Milica B. Simović, Katarina M. Banjanac, Dejan I. Bezbradica

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One of the fundamental requirements for overall human well-being is a stable and balanced microbiome. Aside from the microorganisms that reside within the body, a large number of microorganisms, especially bacteria, swarming the human skin is in homeostasis with the host and represents a skin microbiota. Even though the immune system of the skin is capable of distinguishing between commensal and potentially harmful transient bacteria, the cutaneous microbial balance can be disrupted under certain circumstances. In that case, a reduction in the skin microbiota diversity, as well as changes in metabolic activity, results in dermal infections and inflammation. Probiotics and prebiotics have the potential to play a significant role in the treatment of these skin disorders. The most common resident bacteria found on the skin, Staphylococcus epidermidis, can act as a potential skin probiotic, contributing to the protection of healthy skin from pathogen colonization, such as Staphylococcus aureus, which is related to atopic dermatitis exacerbation. However, as it is difficult to meet regulations in cosmetic products, another therapy approach could be topical prebiotic supplementation of the skin microbiota. In recent research, polyphenols are attracting scientists' interest as biomolecules with possible prebiotic effects on the skin microbiota. This research aimed to determine how herbal extracts rich in different polyphenolic compounds (lemon balm, St. John's wort, coltsfoot, pine needle, and yarrow) affected the growth of S. epidermidis and S. aureus. The first part of the study involved screening plants to determine if they could be regarded as probable candidates to be skin prebiotics. The effect of each plant on bacterial growth was examined by supplementing the nutrient medium with their extracts and comparing it with control samples (without extract). The results obtained after 24 h of incubation showed that all tested extracts influenced the growth of the examined bacteria to some extent. Since lemon balm and St. John's wort extracts displayed bactericidal activity against S. epidermidis, whereas coltsfoot inhibited both bacteria equally, they were not explored further. On the other hand, pine needles and yarrow extract led to an increase in S. epidermidis/S. aureus ratio, making them prospective candidates to be used as skin prebiotics. By examining the prebiotic effect of two extracts at different concentrations, it was revealed that, in the case of yarrow, 0.1% of extract dry matter in the fermentation medium was optimal, while for the pine needle extract, a concentration of 0.05% was preferred, since it selectively stimulated S. epidermidis growth and inhibited S. aureus proliferation. Additionally, the total polyphenols and flavonoid content of the two extracts were determined, revealing different concentrations and polyphenol profiles. Since yarrow and pine extracts affected the growth of skin bacteria in a dose-dependent manner, by carefully selecting the quantities of these extracts, and thus polyphenols content, it is possible to achieve desirable alterations of skin microbiota composition, which may be suitable for the treatment of atopic dermatitis.

Keywords: herbal extracts, polyphenols, skin microbiota, skin prebiotics

Procedia PDF Downloads 155

Authors: Waled A. Dawoud, Abdelazim M. Negm, Nasser M. Saleh

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Offshore piling generates underwater sound at level high enough to cause physical damage or hearing impairment to the marine mammals. Several methods can be used to mitigate the effect of underwater noise from offshore pile driving on marine mammals which can be divided into three main approaches. The first approach is to keep the mammal out of the high-risk area by using aversive sound waves produced by acoustic mitigation devices such as playing-back of mammal's natural predator vocalization, alarm or distress sounds, and anthropogenic sound. The second approach is to reduce the amount of underwater noise from pile driving using noise mitigation techniques such as bubble curtains, isolation casing, and hydro-sound dampers. The third approach is to eliminate the overlap of underwater waves by using prolonged construction process. To investigate the effectiveness of different noise mitigation methods; a pile driven with 235 kJ rated energy diesel hammer near Jeddah Coast, Kingdom of Saudi Arabia was used. Using empirical sound exposure model based on Red Sea characteristics and limits of National Oceanic and Atmospheric Administration; it was found that the aversive sound waves should extend to 1.8 km around the pile location. Bubble curtains can reduce the behavioral disturbance area up to 28%; temporary threshold shift up to 36%; permanent threshold shift up to 50%; and physical injury up to 70%. Isolation casing can reduce the behavioral disturbance range up to 12%; temporary threshold shift up to 21%; permanent threshold shift up to 29%; and physical injury up to 46%. Hydro-sound dampers efficiency depends mainly on the used technology and it can reduce the behavioral disturbance range from 10% to 33%; temporary threshold shift from 18% to 25%; permanent threshold shift from 32% to 50%; and physical injury from 46% to 60%. To prolong the construction process, it was found that the single pile construction, use of soft start, and keep time between two successive hammer strikes more than 3 seconds are the most effective techniques.

Keywords: offshore pile driving, sound propagation models, noise effects on marine mammals, Underwater noise mitigation

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Authors: Geisa Lima Mesquita Zambrosi, Maisa Ciampi Guillardi, Flávia Rodrigues Patrício, Oliveiro Guerreiro Filho

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Certified agricultural products are important components of the food industry. However, certifiers have been expanding the list of restricted or prohibited pesticides, limiting the options of products for phytosanitary control of plant diseases, but without offering alternatives to the farmers. Soybean and coffee leaf rust, brown eye spots, and Phoma leaf spots are the main fungal diseases that pose a serious threat to soybean and coffee cultivation worldwide. In conventional farming systems, these diseases are controlled by using synthetic fungicides, which, in addition to intensifying the occurrence of fungal resistance, are highly toxic to the environment, farmers, and consumers. In organic, agroecological, or regenerative farming systems, product options for plant protection are limited, being available only copper-based compounds, and biodefensivesornon-standard homemade products. Therefore, there is a growing demand for effective bioprotectors with low environmental impact for adoption in more sustainable agricultural systems. Then, to contribute to covering such a gap, we have developed a compound based on plant extracts and metallic elements for foliar application. This product has both biostimulant and bioprotective action, which promotes sustainable disease control, increases productivity as well as reduces damage to the environment. The product's components have complementary mechanisms that promote protection against the disease by directly acting on the pathogens and activating the plant's natural defense system. The protective ability of the product against three coffee diseases (coffee leaf rust, brown eye spot, and Phoma leaf spot) and against soybean rust disease was evaluated, in addition to its ability to promote plant growth. Our goal is to offer an effective alternative to control the main coffee fungal diseases and soybean fungal diseases, with a biostimulant effect and low toxicity. The proposed product can also be part of the integrated management of coffee and soybean diseases in conventional farming associated with chemical and biological pesticides, offering the market a sustainable coffee and soybean with high added value and low residue content. Experiments were carried out under controlled conditions to evaluate the effectiveness of the product in controlling rust, phoma, and cercosporiosis in comparison to control-inoculated plants that did not receive the product. The in vitro and in vivo effects of the product on the pathogen were evaluated using light microscopy and scanning electron microscopy, respectively. The fungistatic action of the product was demonstrated by a reduction of 85% and 95% in spore germination and disease symptoms severity on the leaves of coffee plants, respectively. The formulation had both a protective effect, acting to prevent infection by coffee leaf rust, and a curative effect, reducing the rust symptoms after its establishment.

Keywords: plant disease, natural fungicide, plant health, sustainability, alternative disease management

Procedia PDF Downloads 20

Authors: Z. Yousefniayejahr, S. Bolognini, A. Bonini, C. Campobasso, N. Poma, F. Vivaldi, M. Di Luca, A. Tavanti, F. Di Francesco

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Pathogenic bacteria represent a threat to healthcare systems and the food industry because their rapid detection remains challenging. Electrochemical biosensors are gaining prominence as a novel technology for the detection of pathogens due to intrinsic features such as low cost, rapid response time, and portability, which make them a valuable alternative to traditional methodologies. These sensors use biorecognition elements that are crucial for the identification of specific bacteria. In this context, bacteriophages are promising tools for their inherent high selectivity towards bacterial hosts, which is of fundamental importance when detecting bacterial pathogens in complex biological samples. In this study, we present the development of a low-cost and portable sensor based on the Zeno phage for the rapid detection of Staphylococcus aureus. Screen-printed gold electrodes functionalized with the Zeno phage were used, and electrochemical impedance spectroscopy was applied to evaluate the change of the charge transfer resistance (Rct) as a result of the interaction with S. aureus MRSA ATCC 43300. The phage-based biosensor showed a linear range from 101 to 104 CFU/mL with a 20-minute response time and a limit of detection (LOD) of 1.2 CFU/mL under physiological conditions. The biosensor’s ability to recognize various strains of staphylococci was also successfully demonstrated in the presence of clinical isolates collected from different geographic areas. Assays using S. epidermidis were also carried out to verify the species-specificity of the phage sensor. We only observed a remarkable change of the Rct in the presence of the target S. aureus bacteria, while no substantial binding to S. epidermidis occurred. This confirmed that the Zeno phage sensor only targets S. aureus species within the genus Staphylococcus. In addition, the biosensor's specificity with respect to other bacterial species, including gram-positive bacteria like Enterococcus faecium and the gram-negative bacterium Pseudomonas aeruginosa, was evaluated, and a non-significant impedimetric signal was observed. Notably, the biosensor successfully identified S. aureus bacterial cells in a complex matrix such as a nasal swab, opening the possibility of its use in a real-case scenario. We diluted different concentrations of S. aureus from 108 to 100 CFU/mL with a ratio of 1:10 in the nasal swap matrices collected from healthy donors. Three different sensors were applied to measure various concentrations of bacteria. Our sensor indicated high selectivity to detect S. aureus in biological matrices compared to time-consuming traditional methods, such as enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and radioimmunoassay (RIA), etc. With the aim to study the possibility to use this biosensor to address the challenge associated to pathogen detection, ongoing research is focused on the assessment of the biosensor’s analytical performances in different biological samples and the discovery of new phage bioreceptors.

Keywords: electrochemical impedance spectroscopy, bacteriophage, biosensor, Staphylococcus aureus

Procedia PDF Downloads 50

Authors: Dmitry Lukyanov, Sergey Shevchenko, Alexander Kukaev

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Modern MEMS gyroscopes have strengthened their position in motion control systems and have led to the creation of tactical grade sensors (better than 15 deg/h). This was achieved by virtue of the success in micro- and nanotechnology development, cooperation among international experts and the experience gained in the mass production of MEMS gyros. This production is knowledge-intensive, often unique and, therefore, difficult to develop, especially due to the use of 3D-technology. The latter is usually associated with manufacturing of inertial masses and their elastic suspension, which determines the vibration and shock resistance of gyros. Today, consumers developing highly dynamic objects or objects working under extreme conditions require the gyro shock resistance of up to 65 000 g and the measurement range of more than 10 000 deg/s. Such characteristics can be achieved by solid-state gyroscopes (SSG) without inertial masses or elastic suspensions, which, for example, can be constructed with molecular kinetics of bulk or surface acoustic waves (SAW). Excellent effectiveness of this sensors production and a high level of structural integration provides basis for increased accuracy, size reduction and significant drop in total production costs. Existing principles of SAW-based sensors are based on the theory of SAW propagation in rotating coordinate systems. A short introduction to the theory of a gyroscopic (Coriolis) effect in SAW is provided in the report. Nowadays more and more applications require passive and wireless sensors. SAW-based gyros provide an opportunity to create one. Several design concepts incorporating reflective delay lines were proposed in recent years, but faced some criticism. Still, the concept is promising and is being of interest in St. Petersburg Electrotechnical University. Several experimental models were developed and tested to find the minimal configuration of a passive and wireless SAW-based gyro. Structural schemes, potential characteristics and known limitations are stated in the report. Special attention is dedicated to a novel method of a FEM modeling with piezoelectric and gyroscopic effects simultaneously taken into account.

Keywords: FEM simulation, gyroscope, OOFELIE, surface acoustic wave, wireless sensing

Procedia PDF Downloads 352

Authors: Anatoly A. Kuznetsov, Pavel G. Berezhko, Sergey M. Kunavin, Eugeny V. Zhilkin, Maxim V. Tsarev, Vyacheslav V. Yaroshenko, Valery V. Mokrushin, Olga Y. Yunchina, Sergey A. Mityashin

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The acoustic emission is caused by short-time propagation of elastic waves that are generated as a result of quick energy release from sources localized inside some material. In particular, the acoustic emission phenomenon lies in the generation of acoustic waves resulted from the reconstruction of material internal structures. This phenomenon is observed at various physicochemical transformations, in particular, at those accompanying hydrogenation processes of metals or intermetallic compounds that make it possible to study parameters of these transformations through recording and analyzing the acoustic signals. It has been known that at the interaction between metals or inter metallides with hydrogen the most intensive acoustic signals are generated as a result of cracking or crumbling of an initial compact powder sample as a result of the change of material crystal structure under hydrogenation. This work is dedicated to the study into changes occurring in metallic titanium samples at their interaction with hydrogen and followed by acoustic emission signals. In this work the subjects for investigation were specimens of metallic titanium in two various initial forms: titanium sponge and fine titanium powder made of this sponge. The kinetic of the interaction of these materials with hydrogen, the acoustic emission signals accompanying hydrogenation processes and the structure of the materials before and after hydrogenation were investigated. It was determined that in both cases interaction of metallic titanium and hydrogen is followed by acoustic emission signals of high amplitude generated on reaching some certain value of the atomic ratio [H]/[Ti] in a solid phase because of metal cracking at a macrolevel. The typical sizes of the cracks are comparable with particle sizes of hydrogenated specimens. The reasons for cracking are internal stresses initiated in a sample due to the increasing volume of a solid phase as a result of changes in a material crystal lattice under hydrogenation. When the titanium powder is used, the atomic ratio [H]/[Ti] in a solid phase corresponding to the maximum amplitude of an acoustic emission signal are, as a rule, higher than when titanium sponge is used.

Keywords: acoustic emission signal, cracking, hydrogenation, titanium specimen

Procedia PDF Downloads 364

Authors: Hung-Yuan Chang, Wen-Lung Chiang, Kuo-Liang Wu, Chen-Tsung Lin

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National Space Organization (NSPO) has completed in 2014 the development of a space-borne GPS receiver, including design, manufacture, comprehensive functional test, environmental qualification test and so on. The main performance of this receiver include 8-meter positioning accuracy, 0.05 m/sec speed-accuracy, the longest 90 seconds of cold start time, and up to 15g high dynamic scenario. The receiver will be integrated in the autonomous FORMOSAT-7 NSPO-Built satellite scheduled to be launched in 2019 to execute pre-defined scientific missions. The flight model of this receiver manufactured in early 2015 will pass comprehensive functional tests and environmental acceptance tests, etc., which are expected to be completed by the end of 2015. The space-borne GPS receiver is a pure software design in which all GPS baseband signal processing are executed by a digital signal processor (DSP), currently only 50% of its throughput being used. In response to the booming global navigation satellite systems, NSPO will gradually expand this receiver to become a multi-mode, multi-band, high-precision navigation receiver, and even a science payload, such as the reflectometry receiver of a global navigation satellite system. The fundamental purpose of this extension study is to port some software algorithms such as signal acquisition and correlation, reused code and large amount of computation load to the FPGA whose processor is responsible for operational control, navigation solution, and orbit propagation and so on. Due to the development and evolution of the FPGA is pretty fast, the new system architecture upgraded via an FPGA should be able to achieve the goal of being a multi-mode, multi-band high-precision navigation receiver, or scientific receiver. Finally, the results of tests show that the new system architecture not only retains the original overall performance, but also sets aside more resources available for future expansion possibility. This paper will explain the detailed DSP/FPGA architecture, development, test results, and the goals of next development stage of this receiver.

Keywords: space-borne, GPS receiver, DSP, FPGA, multi-mode multi-band

Procedia PDF Downloads 350

Authors: Zatovska T. V., Nesterova N. V., Baranova G. V., Zagorodnya S. D.

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In recent years, biosensor technologies based on the phenomenon of surface plasmon resonance (SPR) are becoming increasingly used in biology and medicine. Their application facilitates exploration in real time progress of binding of biomolecules and identification of agents that specifically interact with biologically active substances immobilized on the biosensor surface (biochips). Special attention is paid to the use of Biosensor analysis in determining the antibody-antigen interaction in the diagnostics of diseases caused by viruses and bacteria. According to WHO, the diseases that are caused by the herpes simplex virus (HSV), take second place (15.8%) after influenza as a cause of death from viral infections. Current diagnostics of HSV infection include PCR and ELISA assays. The latter allows determination the degree of immune response to viral infection and respective stages of its progress. In this regard, the searches for new and available diagnostic methods are very important. This work was aimed to develop Biosensor chip for detection of specific antibodies to HSV-1 in the human blood serum. The proteins of HSV1 (strain US) were used as antigens. The viral particles were accumulated in cell culture MDBK and purified by differential centrifugation in cesium chloride density gradient. Analysis of the HSV1 proteins was performed by polyacrylamide gel electrophoresis and ELISA. The protein concentration was measured using De Novix DS-11 spectrophotometer. The device for detection of antigen-antibody interactions was an optoelectronic two-channel spectrometer ‘Plasmon-6’, using the SPR phenomenon in the Krechman optical configuration. It was developed at the Lashkarev Institute of Semiconductor Physics of NASU. The used carrier was a glass plate covered with 45 nm gold film. Screening of human blood serums was performed using the test system ‘HSV-1 IgG ELISA’ (GenWay, USA). Development of Biosensor chip included optimization of conditions of viral antigen sorption and analysis steps. For immobilization of viral proteins 0.2% solution of Dextran 17, 200 (Sigma, USA) was used. Sorption of antigen took place at 4-8°C within 18-24 hours. After washing of chip, three times with citrate buffer (pH 5,0) 1% solution of BSA was applied to block the sites not occupied by viral antigen. It was found direct dependence between the amount of immobilized HSV1 antigen and SPR response. Using obtained biochips, panels of 25 positive and 10 negative for the content of antibodies to HSV-1 human sera were analyzed. The average value of SPR response was 185 a.s. for negative sera and from 312 to. 1264 a.s. for positive sera. It was shown that SPR data were agreed with ELISA results in 96% of samples proving the great potential of SPR in such researches. It was investigated the possibility of biochip regeneration and it was shown that application of 10 mM NaOH solution leads to rupture of intermolecular bonds. This allows reuse the chip several times. Thus, in this study biosensor chip for detection of specific antibodies to HSV1 was successfully developed expanding a range of diagnostic methods for this pathogen.

Keywords: biochip, herpes virus, SPR

Procedia PDF Downloads 407

Authors: Hasan Demiroğlu, Uğur Avcıbaşı, Serhan Sakarya, Perihan Ünak

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Implanted devices are progressively practiced in innovative medicine to relieve pain or improve a compromised function. Implant-associated infections represent an emerging complication, caused by organisms which adhere to the implant surface and grow embedded in a protective extracellular polymeric matrix, known as a biofilm. In addition, the microorganisms within biofilms enter a stationary growth phase and become phenotypically resistant to most antimicrobials, frequently causing treatment failure. In such cases, surgical removal of the implant is often required, causing high morbidity and substantial healthcare costs. Staphylococcus aureus is the most common pathogen causing implant-associated infections. Successful treatment of these infections includes early surgical intervention and antimicrobial treatment with bactericidal drugs that also act on the surface-adhering microorganisms. Linezolid is a promising anti-microbial with ant-staphylococcal activity, used for the treatment of MRSA infections. Linezolid is a synthetic antimicrobial and member of oxazolidinoni group, with a bacteriostatic or bactericidal dose-dependent antimicrobial mechanism against gram-positive bacteria. Intensive use of antibiotics, have emerged multi-resistant organisms over the years and major problems have begun to be experienced in the treatment of infections occurred with them. While new drugs have been developed worldwide, on the other hand infections formed with microorganisms which gained resistance against these drugs were reported and the scale of the problem increases gradually. Scientific studies about the production of bacterial biofilm increased in recent years. For this purpose, we investigated the activity of Lin, Lin radiolabeled with 131I (131I-Lin) and cold iodinated Lin (127I-Lin) against clinical strains of Staphylococcus aureus DSM 4910 in biofilm. In the first stage, radio and cold labeling studies were performed. Quality-control studies of Lin and iodo (radio and cold) Lin derivatives were carried out by using TLC (Thin Layer Radiochromatography) and HPLC (High Pressure Liquid Chromatography). In this context, it was found that the binding yield was obtained to be about 86±2 % for 131I-Lin. The minimal inhibitory concentration (MIC) of Lin, 127I-Lin and 131I-Lin for Staphylococcus aureus DSM 4910 strain were found to be 1µg/mL. In time-kill studies of Lin, 127I-Lin and 131I-Lin were producing ≥ 3 log10 decreases in viable counts (cfu/ml) within 6 h at 2 and 4 fold of MIC respectively. No viable bacteria were observed within the 24 h of the experiments. Biofilm eradication of S. aureus started with 64 µg/mL of Lin, 127I-Lin and 131I-Lin, and OD630 was 0.507±0.0.092, 0.589±0.058 and 0.266±0.047, respectively. The media control of biofilm producing Staphylococcus was 1.675±0,01 (OD630). 131I and 127I did not have any effects on biofilms. Lin and 127I-Lin were found less effectively than 131I-Lin at killing cells in biofilm and biofilm eradication. Our results demonstrate that the 131I-Lin have potent anti-biofilm activity against S. aureus compare to Lin, 127I-Lin and media control. This is suggested that, 131I may have harmful effect on biofilm structure.

Keywords: iodine-131, linezolid, radiolabeling, slime layer, Staphylococcus

Procedia PDF Downloads 545

Authors: Elachouri Mostafa, Fakchich Jamila, Lazaar Jamila, Elmadmad Mohammed, Marhom Mostafa

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Of course, there has been a great growth in scientific information about medicinal plants in recent decades, but in many ways this has proved poor compensation, because such information is accessible, in practice, only to a very few people and anyway, rather little of it is relevant to problems of management and utilization, as encountered in the field. Active compounds are used in most traditional medicines and play an important role in advancing sustainable rural livelihoods through their conservation, cultivation, propagation, marketing and commercialization. Medicinal herbs are great resources for various pharmaceutical compounds and urgent measures are required to protect these plant species from their natural destruction and disappearance. Indeed, there is a real danger of indigenous Arab medicinal practices and knowledge disappearing altogether, further weakening traditional Arab culture and creating more insecurity, as well as forsaking a resource of inestimable economic and health care importance. As scientific approach, the ethnopharmacological investigation remains the principal way to improve, evaluate, and increase the odds of finding of biologically active compounds derived from medicinal plants. As developing country, belonging to the Mediterranean basin, Morocco country is endowed with resources of medicinal and aromatic plants. These plants have been used over the millennia for human welfare, even today. Besides, Morocco has a large plant biodiversity, in fact, its medicinal flora account more than 4200 species growing on various bioclimatic zones from subhumide to arid and Saharan. Nevertheless, the human and animal pressure resulting from the increase of rural population needs has led to degradation of this patrimony. In this paper, we focus our attention on ethnopharmacological studies carried out in Morocco. The goal of this work is to clarify the importance of herbs as platform for drugs discovery and further development, to highlight the importance of ethnopharmacological study as approach on discovery of natural products in the health care field, and to discuss the limit of ethnopharmacological investigation of drug discovery in Morocco.

Keywords: Morocco, medicinal plants, ethnopharmacology, natural products, drug-discovery

Procedia PDF Downloads 298

Authors: Md. Samsuddin Ansari, Ashish Arora

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Every year 10 million people fall ill with one of the oldest diseases known as tuberculosis, caused by Mycobacterium tuberculosis. The success of M. tuberculosis as a pathogen is because of its ability to persist in host tissues. Multidrug resistance (MDR) mycobacteria cases increase every day, which is associated with efflux pumps controlled at the level of transcription. The transcription regulators of MDR transporters in bacteria belong to one of the following four regulatory protein families: AraC, MarR, MerR, and TetR. Phenolic acid decarboxylase repressor (PadR), like a family of transcription regulators, is closely related to the MarR family. Phenolic acid decarboxylase repressor (PadR) was first identified as a transcription factor involved in the regulation of phenolic acid stress response in various microorganisms (including Mycobacterium tuberculosis H37Rv). Recently research has shown that the PadR family transcription factors are global, multifunction transcription regulators. Rv0047c is a PadR subfamily-1 protein. We are exploring the biophysical and structural characterization of Rv0047c. The Rv0047 gene was amplified by PCR using the primers containing EcoRI and HindIII restriction enzyme sites cloned in pET-NH6 vector and overexpressed in DH5α and BL21 (λDE3) cells of E. coli following purification with Ni2+-NTA column and size exclusion chromatography. We did DSC to know the thermal stability; the Tm (transition temperature) of protein is 55.29ºC, and ΔH (enthalpy change) of 6.92 kcal/mol. Circular dichroism to know the secondary structure and conformation and fluorescence spectroscopy for tertiary structure study of protein. To understand the effect of pH on the structure, function, and stability of Rv0047c we employed spectroscopy techniques such as circular dichroism, fluorescence, and absorbance measurements in a wide range of pH (from pH-2.0 to pH-12). At low and high pH, it shows drastic changes in the secondary and tertiary structure of the protein. EMSA studies showed the specific binding of Rv0047c with its own 30-bp promoter region. To determine the effect of complex formation on the secondary structure of Rv0047c, we examined the CD spectra of the complex of Rv0047c with promoter DNA of rv0047. The functional role of Rv0047c was characterized by over-expressing the Rv0047c gene under the control of hsp60 promoter in Mycobacterium tuberculosis H37Rv. We have predicted the three-dimensional structure of Rv0047c using the Swiss Model and Modeller, with validity checked by the Ramachandra plot. We did molecular docking of Rv0047c with dnaA, through PatchDock following refinement through FireDock. Through this, it is possible to easily identify the binding hot-stop of the receptor molecule with that of the ligand, the nature of the interface itself, and the conformational change undergone by the protein pattern. We are using X-crystallography to unravel the structure of Rv0047c. Overall the studies show that Rv0047c may have transcription regulation along with providing an insight into the activity of Rv0047c in the pH range of subcellular environment and helps to understand the protein-protein interaction, a novel target to kill dormant bacteria and potential strategy for tuberculosis control.

Keywords: mycobacterium tuberculosis, phenolic acid decarboxylase repressor, Rv0047c, Circular dichroism, fluorescence spectroscopy, docking, protein-protein interaction

Procedia PDF Downloads 98

Authors: Ofonime Ogba, Augustine Akpan

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Objectives and Goals: The study was aimed at determining pathogenic microbial carriage on the external body parts of Camponotus consobrinus which is also known as the banded sugar ant because of its liking for sugar and sweet food. The level of pathogenic microbial carriage of Camponotus consobrinus in association to the environment in which they have been collected is not known. Methods: The ants were purposively collected from four locations including the kitchens, bedroom of various homes, food shops, and bakeries. The sample collection took place within the hours of 6:30 pm to 11:00 pm. The ants were trapped in transparent plastic containers of which sugar, pineapple peels, sugar cane and soft drinks were used as bait. The ants were removed with a sterile spatula and put in 10mls of peptone water in sterile universal bottles. The containers were vigorously shaken to wash the external surface of the ant. It was left overnight and transported to the Microbiology Laboratory, University of Calabar Teaching Hospital for analysis. The overnight peptone broths were inoculated on Chocolate agar, Blood agar, Cystine Lactose Electrolyte-Deficient agar (CLED) and Sabouraud dextrose agar. Incubation was done aerobically and in a carbon dioxide jar for 24 to 48 hours at 37°C. Isolates were identified based on colonial characteristics, Gram staining, and biochemical tests. Results: Out of the 250 Camponotus consobrinus caught for the study, 90(36.0%) were caught in the kitchen, 75(30.0%) in the bedrooms 40(16.0%) in the bakery while 45(18.0%) were caught in the shops. A total of 82.0% prevalence of different microbial isolates was associated with the ants. The kitchen had the highest number of isolates 75(36.6%) followed by the bedroom 55(26.8%) while the bakery recorded the lowest number of isolates 35(17.1%). The profile of micro-organisms associated with Camponotus consobrinus was Escherichia coli 73(30.0%), Morganella morganii 45(18.0%), Candida species 25(10.0%), Serratia marcescens 10(4.0%) and Citrobacter freundii 10(4.0%). Conclusion: Most of the Camponotus consobrinus examined in the four locations harboured potential pathogens. The presence of ants in homes and shops can facilitate the propagation and spread of pathogenic microorganisms. Therefore, the development of basic preventive measures and the control of ants must be taken seriously.

Keywords: Camponotus consobrinus, potential pathogens, microbial isolates, spread

Procedia PDF Downloads 143

Authors: Parul Kulshreshtha, Subrata Sinha, Rakesh Bhatnagar

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This study was conducted by taking B. anthracis as a model pathogen. On infecting a host, B. anthracis secretes three proteins, namely, protective antigen (PA, 83kDa), edema factor (EF, 89 kDa) and lethal factor (LF, 90 kDa). These three proteins are the components of two anthrax toxins. PA binds to the cell surface receptors, namely, tumor endothelial marker (TEM) 8 and capillary morphogenesis protein (CMG) 2. TEM8 and CMG2 interact with LDL-receptor related protein (LRP) 6 for endocytosis of EF and LF. On entering the cell, EF acts as a calmodulin-dependent adenylate cyclase that causes a prolonged increase of cytosolic cyclic adenosine monophosphate (cAMP). LF is a metalloprotease that cleaves most isoforms of mitogen-activated protein kinase kinases (MAPKK/MEK) close to their N-terminus. By secreting these two toxins, B.anthracis ascertains death of the host. Once the systemic levels of the toxins rise, antibiotics alone cannot save the host. Therefore, toxin-specific inhibitors have to be developed. In this wake, monoclonal antibodies have been developed for the neutralization of toxic effects of anthrax toxins. We created hybridomas by using spleen of mice that were actively immunized with rLFn (recombinant N-terminal domain of lethal factor of B. anthracis) to obtain anti-toxin antibodies. Later on, separate group of mice were immunized with rLFn to obtain a polyclonal control for passive immunization studies of monoclonal antibodies. This led to the identification of one cohort of rLFn-immunized mice that harboured disease-enhancing polyclonal antibodies. At the same time, the monoclonal antibodies from all the hybridomas were being tested. Two hybridomas secreted monoclonal antibodies (H8 and H10) that were cross-reactive with EF (edema factor) and LF (lethal factor), while the other two hybridomas secreted LF-specific antibodies (H7 and H11). The protective efficacy of H7, H8, H10 and H11 was investigated. H7, H8 and H10 were found to be protective. H11 was found to have disease enhancing characteristics in-vitro and in mouse model of challenge with B. anthracis. In this study the disease enhancing character of H11 monoclonal antibody and anti-rLFn polyclonal sera was investigated. Combination of H11 with protective monoclonal antibodies (H8 and H10) reduced its disease enhancing nature both in-vitro and in-vivo. But combination of H11 with LETscFv (an scFv with VH and VL identical to H10 but lacking Fc region) could not abrogate the disease-enhancing character of H11 mAb. Therefore it was concluded that for suppression of disease enhancement, Fc portion was absolutely essential for interaction of H10 with H11. Our study indicates that the protective potential of an antibody depends equally on its idiotype/ antigen specificity and its isotype. A number of monoclonal and engineered antibodies are being explored as immunotherapeutics but it is absolutely essential to characterize each one for their individual and combined protective potential. Although new in the sphere of toxin-based diseases, it is extremely important to characterize the disease-enhancing nature of polyclonal as well as monoclonal antibodies. This is because several anti-viral therapeutics and vaccines have failed in the face of this phenomenon. The passive –immunotherapy thus needs to be well formulated to avoid any contraindications.

Keywords: immunotherapy, polyclonal, monoclonal, antibody-dependent disease enhancement

Procedia PDF Downloads 369

Authors: Eleonora Di Salvo, Antonio Panebianco, Graziella Ziino

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Background: Vibrio parahaemolyticus is a human pathogen that is widely distributed in marine environments. It is frequently isolated from raw seafood, particularly shellfish. Consumption of raw or undercooked seafood contaminated with V. parahaemolyticus may lead to acute gastroenteritis. Vibrio spp. has excellent resistance to low temperatures so it can be found in frozen products for a long time. Recently, the viable but non-culturable state (VBNC) of bacteria has attracted great attention, and more than 85 species of bacteria have been demonstrated to be capable of entering this state. VBNC cells cannot grow in conventional culture medium but are viable and maintain metabolic activity, which may constitute an unrecognized source of food contamination and infection. Also V. parahaemolyticus could exist in VBNC state under nutrient starvation or low-temperature conditions. Aim: The aim of the present study was to optimize methods and investigate V. parahaemolyticus VBNC cells and their presence in frozen bivalve molluscs, regularly marketed. Materials and Methods: propidium monoazide (PMA) was integrated with real-time polymerase chain reaction (qPCR) targeting the tl gene to detect and quantify V. parahaemolyticus in the VBNC state. PMA-qPCR resulted highly specific to V. parahaemolyticus with a limit of detection (LOD) of 10-1 log CFU/mL in pure bacterial culture. A standard curve for V. parahaemolyticus cell concentrations was established with the correlation coefficient of 0.9999 at the linear range of 1.0 to 8.0 log CFU/mL. A total of 77 samples of frozen bivalve molluscs (35 mussels; 42 clams) were subsequently subjected to the qualitative (on alkaline phosphate buffer solution) and quantitative research of V. parahaemolyticus on thiosulfate-citrate-bile salts-sucrose (TCBS) agar (DIFCO) NaCl 2.5%, and incubation at 30°C for 24-48 hours. Real-time PCR was conducted on homogenate samples, in duplicate, with and without propidium monoazide (PMA) dye, and exposed for 45 min under halogen lights (650 W). Total DNA was extracted from cell suspension in homogenate samples according to bolliture protocol. The Real-time PCR was conducted with species-specific primers for V. parahaemolitycus. The RT-PCR was performed in a final volume of 20 µL, containing 10 µL of SYBR Green Mixture (Applied Biosystems), 2 µL of template DNA, 2 µL of each primer (final concentration 0.6 mM), and H2O 4 µL. The qPCR was carried out on CFX96 TouchTM (Bio-Rad, USA). Results: All samples were negative both to the quantitative and qualitative detection of V. parahaemolyticus by the classical culturing technique. The PMA-qPCR let us individuating VBNC V. parahaemolyticus in the 20,78% of the samples evaluated with a value between the Log 10-1 and Log 10-3 CFU/g. Only clams samples were positive for PMA-qPCR detection. Conclusion: The present research is the first evaluating PMA-qPCR assay for detection of VBNC V. parahaemolyticus in bivalve molluscs samples, and the used method was applicable to the rapid control of marketed bivalve molluscs. We strongly recommend to use of PMA-qPCR in order to identify VBNC forms, undetectable by the classic microbiological methods. A precise knowledge of the V.parahaemolyticus in a VBNC form is fundamental for the correct risk assessment not only in bivalve molluscs but also in other seafood.

Keywords: food safety, frozen bivalve molluscs, PMA dye, Real-time PCR, VBNC state, Vibrio parahaemolyticus

Procedia PDF Downloads 119

Authors: M. Amparo Nunez-Andres, Nieves Lantada

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One of the most harmful geological risks is rockfalls. They cause both economic lost, damaged in buildings and infrastructures, and personal ones. Therefore, in order to estimate the risk of the exposed elements, it is necessary to know the mechanism of this kind of events, since the characteristics of the rock walls, to the propagation of fragments generated by the initial detached rock mass. In the framework of the research RockModels project, several inventories of rockfalls were carried out along the northeast of the Spanish peninsula and the Mallorca island. These inventories have general information about the events, although the important fact is that they contained detailed information about fragmentation. Specifically, the IBSD (Insitu Block Size Distribution) is obtained by photogrammetry from drone or TLS (Terrestrial Laser Scanner) and the RBSD (Rock Block Size Distribution) from the volume of the fragment in the deposit measured by hand. In order to share all this information with other scientists, engineers, members of civil protection, and stakeholders, it is necessary a platform accessible from the internet and following interoperable standards. In all the process, open-software have been used: PostGIS 2.1., Geoserver, and OpenLayers library. In the first step, a spatial database was implemented to manage all the information. We have used the data specifications of INSPIRE for natural risks adding specific and detailed data about fragmentation distribution. The next step was to develop a WMS with Geoserver. A previous phase was the creation of several views in PostGIS to show the information at different scales of visualization and with different degrees of detail. In the first view, the sites are identified with a point, and basic information about the rockfall event is facilitated. In the next level of zoom, at medium scale, the convex hull of the rockfall appears with its real shape and the source of the event and fragments are represented by symbols. The queries at this level offer a major detail about the movement. Eventually, the third level shows all elements: deposit, source, and blocks, in their real size, if it is possible, and in their real localization. The last task was the publication of all information in a web mapping site (www.rockdb.upc.edu) with data classified by levels using libraries in JavaScript as OpenLayers.

Keywords: geological risk, web mapping, WMS, rockfalls

Procedia PDF Downloads 144

Authors: Baptiste Vallet-Simond, Léopold V. B. Diop, Olivier Isnard

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The antiferromagnetic itinerant-electron compound LaFe₁₂B₆ occupies a special place among rare-earth iron-rich intermetallic; it presents exotic magnetic and physical properties. The unusual amplitude-modulated spin configuration defined by a propagation vector k = (¼, ¼, ¼), remarkably weak Fe magnetic moment (0.43 μB) in the antiferromagnetic ground state, especially low magnetic ordering temperature TN = 36 K for an Fe-rich phase, a multicritical point in the complex magnetic phase diagram, both normal and inverse magnetocaloric effects, and huge hydrostatic pressure effects can be highlighted as the most relevant. Both antiferromagnetic (AFM) and paramagnetic (PM) states can be transformed into the ferromagnetic (FM) state via a field-induced first-order metamagnetic transition. Of particular interest is the low-temperature magnetization process. This process is discontinuous and evolves unexpected huge metamagnetic transitions consisting of a succession of steep magnetization jumps separated by plateaus, giving rise to an unusual avalanche-like behavior. The metamagnetic transition is accompanied by giant magnetoresistance and large magnetostriction. In the present work, we report on the intrinsic magnetic properties of the La₁₋ₓPrₓFe₁₂B₆ series of compounds exhibiting sharp metamagnetic transitions. The study of the structural, magnetic, magneto-transport, and magnetostrictive properties of the La₁₋ₓPrₓFe₁₂B₆ system was performed by combining a wide variety of measurement techniques. Magnetic measurements were performed up to µ0H = 10 T. It was found that the proportion of Pr had a strong influence on the magnetic properties of this series of compounds. At x=0.05, the ground state at 2K is that of an antiferromagnet, but the critical transition field Hc has been lowered from Hc = 6T at x = 0 to Hc = 2.5 Tat x=0.05. And starting from x=0.10, the ground state of this series of compounds is a coexistence of AFM and FM parts. At x=0.30, the AFM order has completely vanished, and only the FM part is left. However, we still observe meta-magnetic transitions at higher temperatures (above 100 K for x=0.30) from the paramagnetic (P) state to a forced FM state. And, of course, such transitions are accompanied by strong magneto-caloric, magnetostrictive, and magnetoresistance effects. The Curie temperatures for the probed compositions going from x=0.05 to x=0.30 were spread over the temperature range of 40 K up to 100 K.

Keywords: metamagnetism, RMB intermetallic, magneto-transport effect, metamagnetic transitions

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Authors: Mohamed Korany, Azza Gazy, Essam Khamis, Marwa Adel, Miranda Fawzy

Abstract:

Background: Two new, simple and specific methods; First, a Zero-crossing first-derivative technique and second, a chemometric-assisted spectrophotometric artificial neural network (ANN) were developed and validated in accordance with ICH guidelines. Both methods were used for the simultaneous estimation of the two closely related antioxidant nutraceuticals ; Coenzyme Q10 (Q) ; also known as Ubidecarenone or Ubiquinone-10, and Vitamin E (E); alpha-tocopherol acetate, in their pharmaceutical binary mixture. Results: For first method: By applying the first derivative, both Q and E were alternatively determined; each at the zero-crossing of the other. The D1 amplitudes of Q and E, at 285 nm and 235 nm respectively, were recorded and correlated to their concentrations. The calibration curve is linear over the concentration range of 10-60 and 5.6-70 μg mL-1 for Q and E, respectively. For second method: ANN (as a multivariate calibration method) was developed and applied for the simultaneous determination of both analytes. A training set (or a concentration set) of 90 different synthetic mixtures containing Q and E, in wide concentration ranges between 0-100 µg/mL and 0-556 µg/mL respectively, were prepared in ethanol. The absorption spectra of the training sets were recorded in the spectral region of 230–300 nm. A Gradient Descend Back Propagation ANN chemometric calibration was computed by relating the concentration sets (x-block) to their corresponding absorption data (y-block). Another set of 45 synthetic mixtures of the two drugs, in defined range, was used to validate the proposed network. Neither chemical separation, preparation stage nor mathematical graphical treatment were required. Conclusions: The proposed methods were successfully applied for the assay of Q and E in laboratory prepared mixtures and combined pharmaceutical tablet with excellent recoveries. The ANN method was superior over the derivative technique as the former determined both drugs in the non-linear experimental conditions. It also offers rapidity, high accuracy, effort and money saving. Moreover, no need for an analyst for its application. Although the ANN technique needed a large training set, it is the method of choice in the routine analysis of Q and E tablet. No interference was observed from common pharmaceutical additives. The results of the two methods were compared together

Keywords: coenzyme Q10, vitamin E, chemometry, quantitative analysis, first derivative spectrophotometry, artificial neural network

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Authors: Kossigan Bernard Dedey, David Grenier, Tiphaine Lucas

Abstract:

Bread dough is often described as a dispersion of gas cells in a continuous gluten/starch matrix. The final bread crumb structure is strongly related to gas cell walls (GCWs) rupture during baking. At the end of proofing and during baking, part of the thinnest GCWs between expanding gas cells is reduced to a gluten film of about the size of a starch granule. When such size is reached gluten and starch granules must be considered as interacting phases in order to account for heterogeneities and appropriately describe GCW rupture. Among experimental investigations carried out to assess GCW rupture, no experimental work was performed to observe the GCW rupture in the baking conditions at GCW scale. In addition, attempts to numerically understand GCW rupture are usually not performed at the GCW scale and often considered GCWs as continuous. The most relevant paper that accounted for heterogeneities dealt with the gluten/starch interactions and their impact on the mechanical behavior of dough film. However, stress concentration in GCW was not discussed. In this study, both experimental and numerical approaches were used to better understand GCW rupture in bread dough during baking. Experimentally, a macro-scope placed in front of a two-chamber device was used to observe the rupture of a real GCW of 200 micrometers in thickness. Special attention was paid in order to mimic baking conditions as far as possible (temperature, gas pressure and moisture). Various differences in pressure between both sides of GCW were applied and different modes of fracture initiation and propagation in GCWs were observed. Numerically, the impact of gluten/starch interactions (cohesion or non-cohesion) and rheological moduli ratio on the mechanical behavior of GCW under unidirectional extension was assessed in 2D/3D. A non-linear viscoelastic and hyperelastic approach was performed to match the finite strain involved in GCW during baking. Stress concentration within GCW was identified. Simulated stresses concentration was discussed at the light of GCW failure observed in the device. The gluten/starch granule interactions and rheological modulus ratio were found to have a great effect on the amount of stress possibly reached in the GCW.

Keywords: dough, experimental, numerical, rupture

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Authors: Ochuko K. Overen, Edson L. Meyer

Abstract:

Uncontrolled heat transfer between the inner and outer space of low-cost housings through the thermal envelope result in indoor thermal discomfort. As a result, an excessive amount of energy is consumed for space heating and cooling. Thermo-optical properties are the ability of paints to reduce the rate of heat transfer through the thermal envelope. The aim of this study is to analyze the thermal performance of a low-cost house with its walls inner surface coated with transparent infrared reflective paint. The thermo-optical properties of the paint were analyzed using Scanning Electron Microscopy/ Energy Dispersive X-ray spectroscopy (SEM/EDX), Fourier Transform Infra-Red (FTIR) and thermal photographic technique. Meteorological indoor and ambient parameters such as; air temperature, relative humidity, solar radiation, wind speed and direction of a low-cost house in Golf-course settlement, South Africa were monitored. The monitoring period covers both winter and summer period before and after coating. The thermal performance of the coated walls was evaluated using time lag and decrement factor. The SEM image shows that the coat is transparent to light. The presence of Al as Al2O and other elements were revealed by the EDX spectrum. Before coating, the average decrement factor of the walls in summer was found to be 0.773 with a corresponding time lag of 1.3 hours. In winter, the average decrement factor and corresponding time lag were 0.467 and 1.6 hours, respectively. After coating, the average decrement factor and corresponding time lag were 0.533 and 2.3 hour, respectively in summer. In winter, an average decrement factor of 1.120 and corresponding time lag of 3 hours was observed. The findings show that the performance of the coats is influenced by the seasons. With a 74% reduction in decrement factor and 1.4 time lag increase in winter, it implies that the coatings have more ability to retain heat within the inner space of the house than preventing heat flow into the house. In conclusion, the results have shown that transparent infrared reflective paint has the ability to reduce the propagation of heat flux through building walls. Hence, it can serve as a remedy to the poor thermal performance of low-cost housings in South Africa.

Keywords: energy efficiency, decrement factor, low-cost housing, paints, rural development, thermal comfort, time lag

Procedia PDF Downloads 264