Search results for: enzyme linked immunosorbent assay

Authors: Litim Tarek, Taamallah Ouahiba

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This work relates to the physico-geometrical aspect of the surface layers of 16NC6 steel having undergone the burnishing treatment by hard steel ball. The results show that the optimal effects of burnishing are closely linked to the shape and the material of the active part of the device as well as to the surface plastic deformation ability of the material to be treated. Thus the roughness is improved by more than 70%, and the consolidation rate is increased by 30%. In addition, modeling of the rational traction curves provides a work hardening coefficient of up to 0.3 in the presence of burnishing.

Keywords: 16NC6 steel, burnishing, hardening, roughness

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Authors: Amir Seyfoori, Ehsan Samiei, Mohsen Akbari

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The use of microtechnology for detection and high yield isolation of circulating tumor cells (CTCs) has shown enormous promise as an indication of clinical metastasis prognosis and cancer treatment monitoring. The Immunomagnetic assay has been also coupled to microtechnology to improve the selectivity and efficiency of the current methods of cancer biomarker isolation. In this way, generation and configuration of the local high gradient magnetic field play essential roles in such assay. Additionally, considering the intrinsic heterogeneity of cancer cells, real-time analysis of isolated cells is necessary to characterize their responses to therapy. Totally, on-chip isolation and monitoring of the specific tumor cells is considered as a pressing need in the way of modified cancer therapy. To address these challenges, we have developed a bi-layer magnetic-based microfluidic chip for enhanced CTC detection and capturing. Micromagnet arrays at the bottom layer of the chip were fabricated using a new method of magnetic nanoparticle paste deposition so that they were arranged at the center of the chain microchannel with the lowest fluid velocity zone. Breast cancer cells labelled with EPCAM-conjugated smart microgels were immobilized on the tip of the micromagnets with greater localized magnetic field and stronger cell-micromagnet interaction. Considering different magnetic nano-powder usage (MnFe2O4 & gamma-Fe2O3) and micromagnet shapes (ellipsoidal & arrow), the capture efficiency of the systems was adjusted while the higher CTC capture efficiency was acquired for MnFe2O4 arrow micromagnet as around 95.5%. As a proof of concept of on-chip tumor cell monitoring, magnetic smart microgels made of thermo-responsive poly N-isopropylacrylamide-co-acrylic acid (PNIPAM-AA) composition were used for both purposes of targeted cell capturing as well as cell monitoring using antibody conjugation and fluorescent dye loading at the same time. In this regard, magnetic microgels were successfully used as cell tracker after isolation process so that by raising the temperature up to 37⁰ C, they released the contained dye and stained the targeted cell just after capturing. This microfluidic device was able to provide a platform for detection, isolation and efficient real-time analysis of specific CTCs in the liquid biopsy of breast cancer patients.

Keywords: circulating tumor cells, microfluidic, immunomagnetic, cell isolation

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Authors: Marwa Elkalla, E. Ali Abdelfadil, Ali. Mohamed. M. Sami, Ali M. Abdel-Monem

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To assess the impact of the blood collection technique on clinical pathology markers and to establish reference intervals, a study was performed using normal, healthy C57BL/6 mice. Both sexes were employed, and they were randomly assigned to different groups depending on the phlebotomy technique used. The blood was drawn in one of four ways: intracardiac (IC), caudal vena cava (VC), caudal vena cava (VC) plus a peritoneal collection of any extravasated blood, or retroorbital phlebotomy (RO). Several serum biochemistries, such as a liver function test, a complete blood count with differentials, and a platelet count, were analysed from the blood and serum samples analysed. Red blood cell count, haemoglobin (p >0.002), hematocrit, alkaline phosphatase, albumin, total protein, and creatinine were all significantly greater in female mice. Platelet counts, specific white blood cell numbers (total, neutrophil, lymphocyte, and eosinophil counts), globulin, amylase, and the BUN/creatinine ratio were all greater in males. The VC approach seemed marginally superior to the IC approach for the characteristics under consideration and was linked to the least variation among both sexes. Transaminase levels showed the greatest variation between study groups. The aspartate aminotransferase (AST) values were linked with decreased fluctuation for the VC approach, but the alanine aminotransferase (ALT) values were similar between the IC and VC groups. There was a lot of diversity and range in transaminase levels between the MC and RO groups. We found that the RO approach, the only one tested that allowed for repeated sample collection, yielded acceptable ALT readings. The findings show that the test results are significantly affected by the phlebotomy technique and that the VC or IC techniques provide the most reliable data. When organising a study and comparing data to reference ranges, the ranges supplied here by collection method and sex can be utilised to determine the best approach to data collection. The authors suggest establishing norms based on the procedures used by each individual researcher in his or her own lab.

Keywords: clinical, pathology, blood, effect

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Authors: Laraib Niaz

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This paper analyzes the ways in which education and economic deprivation are linked to terrorism in Pakistan using data for terrorist incidents from the Global Terrorism Database (GTD). It employs the technique of negative binomial regression for the years between 1990 and 2013, presenting evidence for a positive association between education and terrorism. Conversely, a negative correlation with economic deprivation is signified in the results. The study highlights the element of radicalization as witnessed in the curriculum and textbooks of public schools as a possible reason for extremism, which in turn may lead to terrorism.

Keywords: education, Pakistan, terrorism, poverty

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Authors: Ricardo Avila, Gabriel Lopes, Vania Vidal, Jose Macedo

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This article offers a approach to the automatic discovery of semantic concepts and links in the domain of Oil Exploration and Production (E&P). Machine learning methods combined with textual pre-processing techniques were used to detect local patterns in texts and, thus, generate new concepts and new semantic links. Even using more specific vocabularies within the oil domain, our approach has achieved satisfactory results, suggesting that the proposal can be applied in other domains and languages, requiring only minor adjustments.

Keywords: semantic links, data mining, linked data, SKOS

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Authors: Mohammad Mahdi Namdari, Hosein Torki

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Security as an abstract concept, has involved human being from the beginning of creation to the present, and certainly to the future. Accordingly, battles, conflicts, challenges, legal proceedings, crimes and all issues related to human kind are associated with this concept. Today by interviewing people about their life, the security of societies and Social crimes are interviewed too. Along with the security as an infrastructure and vital concept, the economy and related issues e.g. welfare, per capita income, total government revenue, export, import and etc. is considered another infrastructure and vital concept. These two vital concepts (Security and Economic) have linked together complexly and significantly. The present study employs analytical-descriptive research method using documents and Statistics of official sources. Discovery and explanation of this mutual connection are comprising a profound and extensive research; so management, development and reform in system and relationships of the scope of this two concepts are complex and difficult. Tourism and its position in today's economy is one of the main pillars of the economy of the 21st century that maybe associate with the security and social crimes more than other pillars. Like all human activities, economy of societies and partially tourism dependent on security especially in the public and social security. On the other hand, the true economic development (generally) and the growth of the tourism industry (dedicated) are a security generating and supporting for it, because a dynamic economic infrastructure prevents the formation of centers of crime and illegal activities by providing a context for socio-economic development for all segments of society in a fair and humane. This relationship is a formula of the complexity between the two concept of economy and security. Police as a revealed or people-oriented organization in the field of security directly has linked with the economy of a community and is very effective In the face of the tourism industry. The relationship between security and national crime index, and economic indicators especially ones related to tourism is confirming above discussion that is notable. According to understanding processes about security and economic as two key and vital concepts are necessary and significant for sovereignty of governments.

Keywords: economic, police, tourism, social security

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Authors: Mahmoud H. Aboul-Ela, Riham R. Mohamed, Magdy W. Sabaa

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Synthesis of cross-linked hydrogels composed of trimethyl chitosan (TMC) and poly(vinyl alcohol) (PVA) in different weight ratios in presence of glutaraldehyde as cross-linking agent. Characterization of the prepared hydrogels was done using FTIR, XRD, SEM and TGA. The prepared hydrogels were investigated as adsorbent materials for some transition metal ions from their aqueous solutions. Moreover, the swell ability of the prepared hydrogels was also investigated in both acidic and alkaline pHs, as well as in simulated body fluid (SBF).

Keywords: trimethyl chitosan, hydrogels, metal uptake, superabsorbent materials

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Authors: Fuad Ameen, Mohamed Moslem, Sarfaraz Hadi

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Twenty-eight fungal isolates belonging to 12 genera were derived from debris, sediment and water samples collected from Avicennia marina stands 25km south of Jeddah city on the Red Sea coast of Saudi Arabia. Eight of these isolates were found to be able to grow in association cellulosic waste materials under in vitro conditions in the absence of any carbon source. Isolates were further tested for their potential to degrade paper and clothes wastes by co-cultivation under aeration on a rotary shaker. These fungi accumulated significantly higher biomass, produced ligninolytic and cellulase enzymes, and liberated larger volumes of CO2. These observations indicated that the selected isolates were able to break down and consume the waste materials.

Keywords: biodegradation, enzyme activity, waste materials, mangrove

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Authors: Morgane Chatard, Clémentine Puech, Nathalie Perek, Frédéric Roche

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Several neurological disorders are linked to repeated hypoxia. The impact of such repeated hypoxic stress, on endothelial cells function of the blood-brain barrier (BBB) is little studied in the literature. Indeed, the study of hypoxic stress in cellular pathways is complex using hypoxia exposure because HIF 1α (factor induced by hypoxia) has a short half life. Our study presents an innovative induction method of repeated hypoxic stress, more reproducible, which allows us to study its impacts on an in vitro contact co-culture BBB model. Repeated hypoxic stress was induced by hydralazine (a mimetic agent of hypoxia pathway) during two hours and repeated during 24 hours. Then, BBB integrity was assessed by permeability measurements (transendothelial electrical resistance and membrane permeability), tight junction protein expressions (cell-ELISA and confocal microscopy) and by studying expression and activity of efflux transporters. First, this study showed that repeated hypoxic stress leads to a BBB’s dysfunction illustrated by a significant increase in permeability. This loss of membrane integrity was linked to a significant decrease of tight junctions’ protein expressions, facilitating a possible transfer of potential cytotoxic compounds in the brain. Secondly, we demonstrated that brain microvascular endothelial cells had set-up defence mechanism. These endothelial cells significantly increased the activity of their efflux transporters which was associated with a significant increase in their expression. In conclusion, repeated hypoxic stress lead to a loss of BBB integrity with a decrease of tight junction proteins. In contrast, endothelial cells increased the expression of their efflux transporters to fight against cytotoxic compounds brain crossing. Unfortunately, enhanced efflux activity could also lead to reducing pharmacological drugs delivering to the brain in such hypoxic conditions.

Keywords: BBB model, efflux transporters, repeated hypoxic stress, tigh junction proteins

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Authors: J. E. Mendes, T. T. de Barros, O. B. G. de Assis, J. D. C. Pessoa

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Chitosan, a natural polysaccharide of β-1,4-linked glucosamine residues, is a biopolymer obtained primarily from the exoskeletons of crustaceans. Interest in polymeric materials increases year by year. Chitosan is one of the most plentiful biomaterials, with a wide range of pharmaceutical, biomedical, industrial and agricultural applications. Chitosan nanoparticles were synthesized via the ionotropic gelation of chitosan with sodium tripolyphosphate (TPP). Two concentrations of chitosan microparticles (0.1 and 0.2%) were synthesized. In this study, it was possible to synthesize and characterize microparticles of chitosan biomaterial and this will be used for future applications in cell anchorage for 3D bioprinting.

Keywords: chitosan microparticles, biomaterial, scaffold, bioprinting

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Authors: Avelyno D'Costa, S. K. Shyama, M. K. Praveen Kumar

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The coast of Goa, India receives constant anthropogenic stress through its major rivers which carry mining rejects of iron and manganese ores from upstream mining sites and petroleum hydrocarbons from shipping and harbor-related activities which put the aquatic fauna such as bivalves at risk. The present study reports the pollution status of the Goan coast by the above xenobiotics employing genotoxicity studies. This is further supplemented by the quantification of total petroleum hydrocarbons (TPHs) and various trace metals (iron, manganese, copper, cadmium, and lead) in gills of the estuarine clam, Meretrix ovum as well as from the surrounding water and sediment, over a two-year sampling period, from January 2013 to December 2014. Bivalves were collected from a probable unpolluted site at Palolem and a probable polluted site at Vasco, based upon the anthropogenic activities at these sites. Genotoxicity was assessed in the gill cells using the comet assay and micronucleus test. The quantity of TPHs and trace metals present in gill tissue, water and sediments were analyzed using spectrofluorometry and atomic absorption spectrophotometry (AAS), respectively. The statistical significance of data was analyzed employing Student’s t-test. The relationship between DNA damage and pollutant concentrations was evaluated using multiple regression analysis. Significant DNA damage was observed in the bivalves collected from Vasco which is a region of high industrial activity. Concentrations of TPHs and trace metals (iron, manganese, and cadmium) were also found to be significantly high in gills of the bivalves collected from Vasco compared to those collected from Palolem. Further, the concentrations of these pollutants were also found to be significantly high in the water and sediments at Vasco compared to that of Palolem. This may be due to the lack of industrial activity at Palolem. A high positive correlation was observed between the pollutant levels and DNA damage in the bivalves collected from Vasco suggesting the genotoxic nature of these pollutants. Further, M. ovum can be used as a bioindicator species for monitoring the level of pollution of the estuarine/coastal regions by TPHs and trace metals.

Keywords: comet assay, metals, micronucleus test, total petroleum Hydrocarbons

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Authors: Rajeev Ravindran, Amit K. Jaiswal

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Waste from the food-processing industry is produced in large amount and contains high levels of lignocellulose. Due to continuous accumulation throughout the year in large quantities, it creates a major environmental problem worldwide. The chemical composition of these wastes (up to 75% of its composition is contributed by polysaccharide) makes it inexpensive raw material for the production of value-added products such as biofuel, bio-solvents, nanocrystalline cellulose and enzymes. In order to use lignocellulose as the raw material for the microbial fermentation, the substrate is subjected to enzymatic treatment, which leads to the release of reducing sugars such as glucose and xylose. However, the inherent properties of lignocellulose such as presence of lignin, pectin, acetyl groups and the presence of crystalline cellulose contribute to recalcitrance. This leads to poor sugar yields upon enzymatic hydrolysis of lignocellulose. A pre-treatment method is generally applied before enzymatic treatment of lignocellulose that essentially removes recalcitrant components in biomass through structural breakdown. Present study is carried out to find out the best pre-treatment method for the maximum liberation of reducing sugars from spent coffee waste (SPW). SPW was subjected to a range of physical, chemical and physico-chemical pre-treatment followed by a sequential, combinatorial pre-treatment strategy is also applied on to attain maximum sugar yield by combining two or more pre-treatments. All the pre-treated samples were analysed for total reducing sugar followed by identification and quantification of individual sugar by HPLC coupled with RI detector. Besides, generation of any inhibitory compounds such furfural, hydroxymethyl furfural (HMF) which can hinder microbial growth and enzyme activity is also monitored. Results showed that ultrasound treatment (31.06 mg/L) proved to be the best pre-treatment method based on total reducing content followed by dilute acid hydrolysis (10.03 mg/L) while galactose was found to be the major monosaccharide present in the pre-treated SPW. Finally, the results obtained from the study were used to design a sequential lignocellulose pre-treatment protocol to decrease the formation of enzyme inhibitors and increase sugar yield on enzymatic hydrolysis by employing cellulase-hemicellulase consortium. Sequential, combinatorial treatment was found better in terms of total reducing yield and low content of the inhibitory compounds formation, which could be due to the fact that this mode of pre-treatment combines several mild treatment methods rather than formulating a single one. It eliminates the need for a detoxification step and potential application in the valorisation of lignocellulosic food waste.

Keywords: lignocellulose, enzymatic hydrolysis, pre-treatment, ultrasound

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Authors: Silvia Gastaldello, Maria Grillo, Luca Tassoni, Claudio Maran, Stefano Balbo

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Antioxidants are nowadays attractive not only for the countless benefits to the human and animal health, but also for the perspective of use as food preservative instead of synthetic chemical molecules. In this study, the radical scavenging activity of six protein extracts from pulse and oleaginous seeds was evaluated. The selected matrices are Pisum sativum (yellow pea from two different origins), Carthamus tinctorius (safflower), Helianthus annuus (sunflower), Lupinus luteus cv Mister (lupin) and Glycine max (soybean), since they are economically interesting for both human and animal nutrition. The seeds were grinded and proteins extracted from 20mg powder with a specific vegetal-extraction kit. Proteins have been quantified through Bradford protocol and scavenging activity was revealed using DPPH assay, based on radical DPPH (2,2-diphenyl-1-picrylhydrazyl) absorbance decrease in the presence of antioxidants molecules. Different concentrations of the protein extract (1, 5, 10, 50, 100, 500 µg/ml) were mixed with DPPH solution (DPPH 0,004% in ethanol 70% v/v). Ascorbic acid was used as a scavenging activity standard reference, at the same six concentrations of protein extracts, while DPPH solution was used as control. Samples and standard were prepared in triplicate and incubated for 30 minutes in dark at room temperature, the absorbance was read at 517nm (ABS30). Average and standard deviation of absorbance values were calculated for each concentration of samples and standard. Statistical analysis using t-students and p-value were performed to assess the statistical significance of the scavenging activity difference between the samples (or standard) and control (ABSctrl). The percentage of antioxidant activity has been calculated using the formula [(ABSctrl-ABS30)/ABSctrl]*100. The obtained results demonstrate that all matrices showed antioxidant activity. Ascorbic acid, used as standard, exhibits a 96% scavenging activity at the concentration of 500 µg/ml. At the same conditions, sunflower, safflower and yellow peas revealed the highest antioxidant performance among the matrices analyzed, with an activity of 74%, 68% and 70% respectively (p < 0.005). Although lupin and soybean exhibit a lower antioxidant activity compared to the other matrices, they showed a percentage of 46 and 36 respectively. All these data suggest the possibility to use undervalued edible matrices as antioxidants source. However, further studies are necessary to investigate a possible synergic effect of several matrices as well as the impact of industrial processes for a large-scale approach.

Keywords: antioxidants, DPPH assay, natural matrices, vegetal proteins

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Authors: Hugo Rodríguez

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Migrations of researchers, scientists, and inventors are a widespread phenomenon in modern times. In some cases, migrants stay linked to research groups in their countries of origin, either out of their own conviction or because of government policies. We examine different linear models of technological development (using the Ordinary Least Squares (OLS) technique) in eight selected countries and find that the collaborations between resident and nonresident patent applicants correlate with different levels of performance of the technological policies in three different scenarios. Therefore, the reinforcement of that link must be considered a powerful tool for technological development.

Keywords: development, collaboration, patents, technology

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Authors: Amal. N. Al-Kuraieef

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Smoli bread was made by supplementation sunflower flour which was prepared from sunflower (Dahr-EL-Haea) gray after hilling and milling, flour was irradiated by two doses (5 and 10 kGy). After that, the ratios of irradiated sunflower flour were 5 and 10%. All samples of samoli bread were examined for organoleptic and biological evaluation. Biological assay (PER, NPU, FE, DC and BV) was carried out on rats fed 5 and 10% irradiated and non-irradiated sunflower Samoli bread. Results obtained showed that, total lipids, cholesterol and triglycerides were reduced comparable, to that of casein. Also, figures of the biological evaluations were higher than those of the control samoli bread and improved its nutritive values.

Keywords: gamma rays, sunflower, samoli bread, cholesterol, lipids, triglycerides

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Authors: Udeshika Yapa Bandara, Chamindri Witharana, Preethi Soysa

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Pomegranate fruits are used in cancer treatment in Ayurveda, Sri Lanka. Due to prevailing therapeutic effects of phytochemicals, this study was focus on anti-cancer properties of the constituents in the parts of Pomegranate fruit. Furthermore, the method of extraction, plays a crucial step of the phytochemical analysis. Therefore, this study was focus on different extraction methods. Five techniques were involved for the peel and the pericarp to evaluate the most effective extraction method; Boiling with electric burner (BL), Sonication (SN), Microwaving (MC), Heating in a 50°C water bath (WB) and Sonication followed by Microwaving (SN-MC). The presence of polyphenolic and flavonoid contents were evaluated to recognize the best extraction method for polyphenols. The total phenolic content was measured spectrophotometrically by Folin-Ciocalteu method and expressed as Gallic Acid Equivalents (w/w% GAE). Total flavonoid content was also determined spectrophotometrically with Aluminium chloride colourimetric assay and expressed as Quercetin Equivalents (w/w % QE). Pomegranate juice was taken as fermented juice (with Saccharomyces bayanus) and fresh juice. Powdered seeds were refluxed, filtered and freeze-dried. 2g of freeze-dried powder of each component was dissolved in 100ml of De-ionized water for extraction. For the comparison of antioxidant activity and total phenol content, the polyphenols were removed by the Polyvinylpolypyrrolidone (PVVP) column and fermented and fresh juice were tested for the 1, 1-diphenyl-2-picrylhydrazil (DPPH) radical scavenging activity, before and after the removal of polyphenols. For the peel samples of Pomegranate fruit, total phenol and flavonoid contents were high in Sonication (SN). In pericarp, total phenol and flavonoid contents were highly exhibited in method of Sonication (SN). A significant difference was observed (P< 0.05) in total phenol and flavonoid contents, between five extraction methods for both peel and pericarp samples. Fermented juice had a greatest polyphenolic and flavonoid contents comparative to fresh juice. After removing polyphenols of fermented juice and fresh juice using Polyvinyl polypyrrolidone (PVVP) column, low antioxidant activity was resulted for DPPH antioxidant activity assay. Seeds had a very low total phenol and flavonoid contents according to the results. Although, Pomegranate peel is the main waste component of the fruit, it has an excellent polyphenolic and flavonoid contents compared to other parts of the fruit, devoid of the method of extraction. Polyphenols play a major role for antioxidant activity.

Keywords: antioxidant activity, flavonoids, polyphenols, pomegranate

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Authors: Henry Memczak, Marc Hovestaedt, Bernhard Ay, Sandra Saenger, Thorsten Wolff, Frank F. Bier

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The influenza viruses cause flu epidemics every year and serious pandemics in larger time intervals. The only cost-effective protection against influenza is vaccination. Due to rapid mutation continuously new subtypes appear, what requires annual reimmunization. For a correct vaccination recommendation, the circulating influenza strains had to be detected promptly and exactly and characterized due to their antigenic properties. During the flu season 2016/17, a wrong vaccination recommendation has been given because of the great time interval between identification of the relevant influenza vaccine strains and outbreak of the flu epidemic during the following winter. Due to such recurring incidents of vaccine mismatches, there is a great need to speed up the process chain from identifying the right vaccine strains to their administration. The monitoring of subtypes as part of this process chain is carried out by national reference laboratories within the WHO Global Influenza Surveillance and Response System (GISRS). To this end, thousands of viruses from patient samples (e.g., throat smears) are isolated and analyzed each year. Currently, this analysis involves complex and time-intensive (several weeks) animal experiments to produce specific hyperimmune sera in ferrets, which are necessary for the determination of the antigen profiles of circulating virus strains. These tests also bear difficulties in standardization and reproducibility, which restricts the significance of the results. To replace this test a peptide-based assay for influenza virus subtyping from corresponding virus samples was developed. The differentiation of the viruses takes place by a set of specifically designed peptidic recognition molecules which interact differently with the different influenza virus subtypes. The differentiation of influenza subtypes is performed by pattern recognition guided by machine learning algorithms, without any animal experiments. Synthetic peptides are immobilized in multiplex format on various platforms (e.g., 96-well microtiter plate, microarray). Afterwards, the viruses are incubated and analyzed comparing different signaling mechanisms and a variety of assay conditions. Differentiation of a range of influenza subtypes, including H1N1, H3N2, H5N1, as well as fine differentiation of single strains within these subtypes is possible using the peptide-based subtyping platform. Thereby, the platform could be capable of replacing the current antigenic characterization of influenza strains using ferret hyperimmune sera.

Keywords: antigenic characterization, influenza-binding peptides, influenza subtyping, influenza surveillance

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Authors: M. R. Vijayakumar, K. Priyanka, Ramoji Kosuru, Lakshmi, Sanjay Singh

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Trans resveratrol (RES) is a non-flavonoid poly-phenolic compound proved for its therapeutic and preventive effect against various types of cancer. However, the practical application of RES in cancer treatment is limited because of its higher dose (up to 7.5 g/day in humans), low biological half life, rapid metabolism and faster elimination in mammals. PEGylated core-shell type lipid polymer hybrid nanoparticles are the novel drug delivery systems for long circulation and improved anti cancer effect of its therapeutic payloads. Therefore, the main objective of this study is to extend the biological half life (long circulation) and improve the therapeutic efficacy of RES through core shell type of nanoparticles. D-α-tocopheryl polyethylene glycol 1000 succinate (vitamin E TPGS), a novel surfactant is applied for the preparation of PEGylated lipid polymer hybrid nanoparticles. The prepared nanoparticles were evaluated by various state of the art techniques such as dynamic light scattering (DLS) technique for particle size and zeta potential, TEM for shape, differential scanning calorimetry (DSC) for interaction analysis and XRD for crystalline changes of drug. Entrapment efficiency and invitro drug release were determined by ultracentrifugation method and dialysis bag method, respectively. Cancer cell viability studies were performed by MTT assay, respectively. Pharmacokinetic studies after i.v administration were performed in sprague dawley rats. The prepared NPs were found to be spherical in shape with smooth surfaces. Particle size and zeta potential of prepared NPs were found to be in the range of 179.2±7.45 to 266.8±9.61 nm and -0.63 to -48.35 mV, respectively. DSC revealed absence of potential interaction. XRD study revealed presence of amorphous form in nanoparticles. Entrapment efficiency was found to be 83.7 % and drug release was found to be in controlled manner. MTT assay showed low MEC and pharmacokinetic studies showed higher AUC of nanoformulaition than its pristine drug. All these studies revealed that the RES loaded PEG modified core-shell type lipid polymer hybrid nanoparticles can be an alternative tool for chemopreventive and therapeutic application of RES in cancer.

Keywords: trans resveratrol, cancer nanotechnology, long circulating nanoparticles, bioavailability enhancement, core shell nanoparticles, lipid polymer hybrid nanoparticles

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Authors: Abdelaaty A. Shahat, Mansour S. Alsaid

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Four Acetylated flavonol glycosides were isolated from Centaurium spicatum (L.) Fritsch (Gentianaceae). Structure elucidation, especially the localization of the acetyl groups, and complete 1H and 13C NMR assignments of these biologically active compounds were carried out using one- and two-dimensional NMR methods, including CNMR, DEPT-135 and DEPT-90 and gradient-assisted experiments such as DQF-COSY, TOCSY, HSQC and HMBC experiments. The antioxidant activities of the new acetylated flavonoid glycosides using DPPH• assay were determined. The compounds tested showed a good DPPH• activity compared with control, but their activity was lower than that of their corresponding aglycone, quercetin.

Keywords: Centaurium spicatum, flavonoids, biological activity, isolation, glycosides

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Authors: Mayuva Youngsabanant, Chanikarn Srinark, Supanyika Sengsai, Soratorn Kerdkriangkrai, Nongnuch Gumlungpat, Mayuree Pumipaiboon

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The follicular fluid proteins of healthy medium size follicles (4-6 mm in diameters) and large size follicles (7-8 mm in diameter) of large white pig ovaries were collected by using sterile technique. They were used for testing the effect on primary in vitro cell culture growth of porcine oviductal epithelial cells (pOEC). Porcine oviductal epithelial cells of luteal phase was culture in M199 and added with 10% fetal calf serum 2.2 mg/mL, NaHCO₃, 0.25 mM pyruvate, 15 µg/mL and 50 µg/mL, gentamycin sulfate at high humidified atmosphere with 5% CO₂ in 95% air atmosphere at 37°C for 96 h before testing. The optimized concentration of pFF of two follicle sizes (at concentration of 2, 4, 20, 40, 200, 400, 500, and 600 µg proteins) in culture medium was observed for 24 h using MTT assay. Results were analyzed with a one-way ANOVA in SPSS statistic. Moreover, pOEC was also studied in morphological characteristic on long-term culture. The results of long-term study revealed that pOEC showed 70-80 percentage of healthy morphology on epithelial-like character and contained 30 percentage of an elongated shape (fibroblast-like morphology) at 4 weeks of culture time. MTT assay reviewed an increase in the percentage of viability of pOEC in 2 treated of follicular fluid groups. Two treatment concentration groups were higher than control group (p < 0.05) but not in positive control group. Interestingly, at 200 µg protein of 2 treated follicular fluid groups were reached the highest cell viability which is higher than a positive control and it is significantly different form control group (P < 0.05). These cells are developed and had fibroblast elongate shape which is longer than the cells in control group and positive control group. This report implies that pFF of medium follicle size at 200 µg proteins and large follicle size at 200 and 500 µg proteins could be optimized concentration for using as a supplement in culture medium to promote cell growth and development instead of growth hormone from fetal calf serum. It could be applied in cell biotechnology researches. Acknowledgements: The project was funded by a grant from Silpakorn University Research and Development Institute (SURDI) and Faculty of Science, Silpakorn University, Thailand.

Keywords: in vitro, porcine follicular fluid protein (pFF), porcine oviductal epithelial cells (pOEC), MTT

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Authors: C. C. Castro, T. Senechal, D. Lahem, A. L. Hantson

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Formaldehyde is one of the most representative volatile organic compounds present in the indoor air of residential units and workplaces. Increased attention has been given to this toxic compound because of its carcinogenic effect in health. Biological or enzymatic transformation is being explored to degrade this pollutant. Pseudomonas putida is a bacteria able to synthesize formaldehyde dehydrogenase, an enzyme known to use formaldehyde as a substrate and transform it into less toxic compounds. The immobilization of bacterial cells in the surface of different supports through spraying or dip-coating is herein proposed. The determination of the enzymatic activity on the coated surfaces was performed as well as the study of its effect on formaldehyde degradation in an isolated chamber. Results show that the incorporation of microbial cells able to synthesize depolluting enzymes can be an innovative, low-cost, effective and environmentally friendly solution for indoor air depollution.

Keywords: cells encapsulation, formaldehyde, formaldehyde dehydrogenase, indoor air depollution

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Authors: Mesfin Angaw Tesfay

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Uridine-5’-diphosphate (UDP)-glucose is one of the most versatile building blocks within the metabolism of prokaryotes and eukaryotes, serving as an activated sugar donor during the glycosylation of natural products. It is formed by the enzyme UDP-glucose pyrophosphorylase (UGPase) using uridine-5′-triphosphate (UTP) and α-d-glucose 1-phosphate as a substrate. Herein, two UGPase genes from Lactobacillus kefiranofaciens ZW3 (LkUGPase) and Rhodococcus wratislaviensis IFP 2016 (RwUGPase) were identified through genome mining approaches. The LkUGPase and RwUGPase have 299 and 306 amino acids, respectively. Both UGPase has the conserved UTP binding site (G-X-G-T-R-X-L-P) and the glucose -1-phosphate binding site (V-E-K-P). The LkUGPase and RwUGPase were cloned in E. coli, and SDS-PAGE analysis showed the expression of both enzymes forming about 36 KDa of protein band after induction. LkUGPase and RwUGPase have an activity of 1549.95 and 671.53 U/mg, respectively. Currently, their kinetic properties are under investigation.

Keywords: UGPase, LkUGPase, RwUGPase, UDP-glucose, glycosylation

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Authors: Andrés Lara Guillén, José M. Soriano Disla, Gemma Castejón Martínez, David Fernández-Gutiérrez

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The worldwide population is exponentially increasing, which causes a rising demand for food, energy and non-renewable resources. These demands must be attended to from a circular economy point of view. Under this approach, the obtention of strategic products from biowaste is crucial for the society to keep the current lifestyle reducing the environmental and social issues linked to the lineal economy. This is the main objective of the VALUEWASTE project. VALUEWASTE is about valorizing urban biowaste into proteins for food and feed and biofertilizers, closing the loop of this waste stream. In order to achieve this objective, the project validates three value chains, which begin with the anaerobic digestion of the biowaste. From the anaerobic digestion, three by-products are obtained: i) methane that is used by microorganisms, which will be transformed into microbial proteins; ii) digestate that is used by black soldier fly, producing insect proteins; and iii) a nutrient-rich effluent, which will be transformed into biofertilizers. VALUEWASTE is an innovative solution, which combines different technologies to valorize entirely the biowaste. However, it is also required to demonstrate that the solution is greener than other traditional technologies (baseline systems). On one hand, the proteins from microorganisms and insects will be compared with other reference protein production systems (gluten, whey and soybean). On the other hand, the biofertilizers will be compared to the production of mineral fertilizers (ammonium sulphate and synthetic struvite). Therefore, the aim of this study is to provide that biowaste valorization can reduce the environmental impacts linked to both traditional proteins manufacturing processes and mineral fertilizers, not only at a pilot-scale but also at an industrial one. In the present study, both baseline system and VALUEWASTE solution are evaluated through the Environmental Life Cycle Assessment (E-LCA). The E-LCA is based on the standards ISO 14040 and 14044. The Environmental Footprint methodology was the one used in this study to evaluate the environmental impacts. The results for the baseline cases show that the food proteins coming from whey have the highest environmental impact on ecosystems compared to the other proteins sources: 7.5 and 15.9 folds higher than soybean and gluten, respectively. Comparing feed soybean and gluten, soybean has an environmental impact on human health 195.1 folds higher. In the case of biofertilizers, synthetic struvite has higher impacts than ammonium sulfate: 15.3 (ecosystems) and 11.8 (human health) fold, respectively. The results shown in the present study will be used as a reference to demonstrate the better environmental performance of the bio-based products obtained through the VALUEWASTE solution. Other originalities that the E-LCA performed in the VALUEWASTE project provides are the diverse direct implications on investment and policies. On one hand, better environmental performance will serve to remove the barriers linked to these kinds of technologies, boosting the investment that is backed by the E-LCA. On the other hand, it will be a germ to design new policies fostering these types of solutions to achieve two of the key targets of the European Community: being self-sustainable and carbon neutral.

Keywords: anaerobic digestion, biofertilizers, circular economy, nutrients recovery

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Authors: Achiraya Jiraprasertwong, Erdogan Gulari, Sumaeth Chavadej

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Among agricultural residues, sugarcane bagasse is one of the most convincing raw materials for the production of bioethanol due to its availability, and low cost through enzymatic hydrolysis and yeast fermentation. A pretreatment step is needed to enhance the enzymatic step. In this study, sugarcane bagasse (SCB), one of the most abundant agricultural residues in Thailand, was pretreated biologically with various microorganisms of white-rot fungus—Phanerochaete sordid (SK 7), Cellulomonas sp. (TISTR 784), and strain A 002 (Bacillus subtilis isolated from Thai higher termites). All samples with various microbial pretreatments were further hydrolyzed enzymatically by a commercial enzyme obtained from Aspergillus niger. The results showed that the pretreatment with the white-rot fungus gave the highest glucose concentration around two-fold higher when compared with the others.

Keywords: sugarcane bagasse, microorganisms, pretreatment, enzymatic hydrolysis

Procedia PDF Downloads 441

Authors: D. ChobfroushKhoei, S. K. Heidari , Sh. Dariadel

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Carbon nanotubes were used in medical sciences especially in drug delivery system and cancer therapy. In this study, we functionalized carboxylated single-wall carbon nanotubes (SWNT-COOH) with 2-en 4-hydroxy cyclo 1-octanon. Synthesized sample was characterized by FT-IR, Raman spectroscopy, SEM, TGA and cellular investigations. The results showed well formation of SWNT-Ester. Cell viability assay results and microscopic observations demonstrated that cancerous cells were killed in the sample. The synthesized sample can be used as a toxic material for cancer therapy.

Keywords: MWNT-COOH, functionalization, phenylisocyanate, phenylisothiocyanate, 1, 4-phenylendiamine, toxicity investigation

Procedia PDF Downloads 448

Authors: Balgabay Sadepovich Maikanov, Laura Tyulegenovna Auteleyeva, Seidenova Simbat Polatbekovna

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In this article reduced nutritional value of the rabbits’ meat at 1, 1 dimethylhydrazine experimental toxicosis is shown. The assay was performed on liquid chromatograph SHIMADZU LC-20 Prominence (Japan) with fluorometric and spectrophotometric detector. This research has revealed that samples of rabbit meat of the experimental group had significant differences from the control group:in amino acids concentration from 1.2% to 9.1%; vitamin concentration from 11.2% to 60.5%, macro – minerals concentration from 17.4% to 78.1% and saturated fatty acids concentration from 17,1% to 34.5%, respectively. The decrease in the chemical composition of rabbits’ meat at 1,1 dimethylhydrazine toxicosis may be due to changes in the internal processes associated with impaired metabolic homeostasis of animals.

Keywords: 1, 1-dimethylhydrazine, metabolic homeostasis, nutritional value, rabbit meat

Procedia PDF Downloads 212

Authors: Elena Maria Scalisi

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The increasing production and the use of TiO₂ nanoparticles (NPs) have inevitably led to their release into the environment, thereby posing a threat to organisms and also for human. Human exposure to TiO₂-NPs may occur during both manufacturing and use. TiO₂-NPs are common in consumer products for dermal application, toothpaste, food colorants, and nutritional supplements, then oral exposure may occur during use of such products. Into the body, TiO₂-NPs thanks to their small size (<100 nm), can, through testicular blood barrier inducing effect on testis and then on male reproductive health. The nanoscale size of TiO₂ increase the surface-to-volume ratio making them more reactive in a cell, then TiO₂ NPs increase their ability to produce reactive oxygen species (ROS). In male germ cells, ROS may have important implications in maintaining the normal functions of mature spermatozoa at physiological levels, moreover, in spermatozoa they are important signaling molecules for their hyperactivation and acrosome reaction. Nevertheless, an excess of ROS by external inputs such as NPs can increased the oxidative stress (OS), which results in damage DNA and apoptosis. The aim of our study has been investigate the impact of TiO₂ NPs on human spermatozoa, evaluating DNA damage and the expression of proteins involved in cell stress. According WHO guidelines 2021, we have exposed human spermatozoa in vitro to TiO₂ NP at concentrations 50 ppm, 100 ppm, 250 ppm, and 500 ppm for 1 hour (at 37°C and CO₂ at 5%). DNA damage was evaluated by Sperm Chromatin Dispersion Test (SCD) and TUNEL assay; moreover, we have evaluated the expression of biomarkers of oxidative stress like Heat Shock Protein 70 (HSP70) and Metallothioneins (MTs). Also, sperm parameters as motility viability have been evaluated. Our results not report a significant reduction in motility of spermatozoa at the end of the exposure. On the contrary, the progressive motility was increased at the highest concentration (500 ppm) and was statistically significant compared to control (p <0.05). Also, viability was not changed by exposure to TiO₂-NPs (p <0.05). However, increased DNA damage was observed at all concentrations, and the TUNEL assay highlighted the presence of single strand breaks in the DNA. The spermatozoa responded to the presence of TiO₂-NPs with the expression of Hsp70, which have a protective function because they allow the maintenance of cellular homeostasis in stressful/ lethal conditions. A positivity for MTs was observed mainly for the concentration of 4 mg/L. Although the biological and physiological function of the metallothionein (MTs) in the male genital organs is unclear, our results highlighted that the MTs expressed by spermatozoa maintain their biological role of detoxification from metals. Our results can give additional information to the data in the literature on the toxicity of TiO₂-NPs and reproduction.

Keywords: human spermatozoa, DNA damage, TiO₂-NPs, biomarkers

Procedia PDF Downloads 143

Authors: Arash Jafari, Somaye Bahrami, Mohammad Hossein Razi Jalali

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Dicrocoeliosis, caused by Dicrocoelium dendriticum is a hepatic parasitic disease of clinical and financial significance in ruminant breeding, which causes direct losses due to condemnation of parasitized livers. The purpose of our study was to assess the effects of natural dicrocoeliosis on the antioxidant defense capability of the liver in sheep. For this purpose, livers of 40 infected sheep with D. dendriticumalong with livers of 20 healthy (control) sheep were collected from animals slaughtered in Khuzestan province, Iran. An increase in malondialdehyde concentrations accompanied by decreased activities of SOD and GPX of infected liver was noticed when com-pared with control values. Our data indicate that through dicrocoeliosis insufficient scavenging of reactive oxygen species takes place and caused oxidative liver damage.

Keywords: Dicrocoelium dendriticum, lipid peroxidation, antioxidant enzyme, liver

Procedia PDF Downloads 664

Authors: Stephanie Newman

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Niemann-Pick Type C disease is a rare, usually fatal lysosomal storage disorder. Although clinically characterized by progressive neurodegeneration, there is also evidence of altered innate immune responses such as neuroinflammation that promote disease progression. We have initiated an investigation into whether phagocytosis, an important innate immune activity and the process by which particles are ingested is defective in NPC. Using an in vitro assay, we have shown that NPC macrophages have a deficiency in the phagocytosis of different particles. We plan to investigate the mechanistic basis for impaired phagocytosis, the contribution that this deficiency makes to disease pathology, and whether therapies that have shown in vivo benefit are able to restore phagocytic activity.

Keywords: Niemann Pick Disease C, phagocytosis, innate immunity, lysosomal storage disorder

Procedia PDF Downloads 391

Authors: Merve Kurt

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How women should behave in public spaces and how they should be dressed was a loaded issues in the Ottoman Empire. They pointed to what kind of state the Ottoman State was. One of such public space was Mesires, promenades. Women's visibility and invisibility, their morals were reflected and linked to the society as a whole. How the public space and private space is defined, what were the lines that separates them, how much blurred these lines were discussed in this paper. Moreover, all these points were strengthened by the primary sources from archives dating to the end of the 19th century.

Keywords: Mesire, Ottoman Empire, Ottoman women, public spaces

Procedia PDF Downloads 229