Search results for: polymerase activity

Authors: Natan C. G. Silva, Carlos A. C. Girao Neto, Marcele M. S. Vasconcelos, Luciana R. B. Goncalves, Maria Valderez P. Rocha

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Galactooligosaccharides (GOS) are sugars with prebiotic function that can be synthesized chemically or enzymatically, and this last one can be promoted by the action of β-galactosidases. In addition to favoring the transgalactosylation reaction to form GOS, these enzymes can also catalyze the hydrolysis of lactose. A highly studied type of GOS is lactulose because it presents therapeutic properties and is a health promoter. Among the different raw materials that can be used to produce lactulose, whey stands out as the main by-product of cheese manufacturing, and its discarded is harmful to the environment due to the residual lactose present. Therefore, its use is a promising alternative to solve this environmental problem. Thus, lactose from whey is hydrolyzed into glucose and galactose by β-galactosidases. However, in order to favor the transgalactosylation reaction, the medium must contain fructose, due this sugar reacts with galactose to produce lactulose. Then, the glucose-isomerase enzyme can be used for this purpose, since it promotes the isomerization of glucose into fructose. In this scenario, the aim of the present work was first to develop β-galactosidase biocatalysts of Kluyveromyces lactis and to apply it in the integrated reactions of hydrolysis, isomerization (with the glucose-isomerase from Streptomyces murinus) and transgalactosylation reaction, using whey as a substrate. The immobilization of β-galactosidase in chitosan previously functionalized with 0.8% glutaraldehyde was evaluated using different enzymatic loads (2, 5, 7, 10, and 12 mg/g). Subsequently, the hydrolysis and transgalactosylation reactions were studied and conducted at 50°C, 120 RPM for 20 minutes. In parallel, the isomerization of glucose into fructose was evaluated under conditions of 70°C, 750 RPM for 90 min. After, the integration of the three processes for the production of lactulose was investigated. Among the evaluated loads, 7 mg/g was chosen because the best activity of the derivative (44.3 U/g) was obtained, being this parameter determinant for the reaction stages. The other parameters of immobilization yield (87.58%) and recovered activity (46.47%) were also satisfactory compared to the other conditions. Regarding the integrated process, 94.96% of lactose was converted, achieving 37.56 g/L and 37.97 g/L of glucose and galactose, respectively. In the isomerization step, conversion of 38.40% of glucose was observed, obtaining a concentration of 12.47 g/L fructose. In the transgalactosylation reaction was produced 13.15 g/L lactulose after 5 min. However, in the integrated process, there was no formation of lactulose, but it was produced other GOS at the same time. The high galactose concentration in the medium probably favored the reaction of synthesis of these other GOS. Therefore, the integrated process proved feasible for possible production of prebiotics. In addition, this process can be economically viable due to the use of an industrial residue as a substrate, but it is necessary a more detailed investigation of the transgalactosilation reaction.

Keywords: beta-galactosidase, glucose-isomerase, galactooligosaccharides, lactulose, whey

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Authors: Taşkın Deniz Yıldız, Mustafa Topaloğlu, Orhan Kural

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The right to operate a fairly long periods of prior periods and after the 3213 Mining Law has been observed to be shortened in Turkey. Permit the realization of business activities (or concession) requested the purchase of the mine operated "found mine" position, as well as the financial and technical capability to have the owner of the right to operate the mines as well as the principle of equality is important in terms of assessing the best way be. In particular, in this context, license fields "negligence" (downsizing) have noted that the current arrangement for all periods. However, in the period after 3213 Mining Act and a permit to operate more effectively within the framework of implementation of negligence is laid down.

Keywords: mining legislation, operation, permit, Turkey

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Authors: Mohinder Kumar

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Sports are considered as the very prominent social institution in almost every society because it reflects the mores, values, and general culture of a society. Sports activity tend to pave the foundation for learning acceptable values and beliefs and for acquiring desirable character traits such as self-discipline, sportsmanship, and an appreciation for hard work, fairness, self-respect, leadership, followership, justice, perseverance, competition, and goal attainment. The present study focuses on ideal ways of socializing youngsters into sports. Influences of some socializing agents (e.g. family, school, community) are reviewed and suggestions made as to how these socializing agents can be oriented and made effective in carrying out functional processes toward target ends.

Keywords: sports, socializing, family, community, society

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Authors: Ahmad Saleem

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Ethereum is the second largest crypto currency in blockchain ecosystem. Along with standard transactions, it supports smart contracts and NFT’s. Current research trends are focused on analyzing the overall structure of the network its growth and behavior. Ethereum addresses are anonymous and can be created on fly. The nature of Ethereum network and addresses make it hard to predict their behavior. The activity period of an ethereum address is not much analyzed. Using machine learning we can make early prediction about the disposability of the address. In this paper we analyzed the lifetime of the addresses. We also identified and predicted the disposable addresses using machine learning models and compared the results.

Keywords: blockchain, Ethereum, cryptocurrency, prediction

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Authors: Chiung-Hua Chan, Fang-Yuan Chang, Li-Chi Huang

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Stroke patients received regular rehabilitation therapy have measurable advancement in muscle strength, balance, control upper and lower physical activity, walking speed and endurance. This study aimed to investigate the relationship between increases in bedside rehabilitation time and the function of activities and daily living (ADL) in stroke patients. The study was quasi-experimental research design and randomized sampling. The researcher collected 12 stroke patients of stroke patients transferred to rehabilitation ward unit of a medical center during 1 January to 31 March 2017. All participants then were assigned to case group and control group. Data collection was through direct observation of assessment ADL of stroke patients by researchers on Day 1. Case group received regular rehabilitation, exercises in increase of bedside rehabilitation schedules exercise programs by ward nurses. Bedside rehabilitation exercise content with physical, functional and linguistic frequency and time, Control group only give routine rehabilitation schedule care. This was a randomized study performed in 12 patients who were stroke patients and transferred to rehabilitation ward unit of a medical center during 1 January to 31 March 2017. First, the researcher explained the purpose and method of the study to the patients or the family members. All participants completed a consent informed before participation. Patients were randomly assigned to a ‘bedside rehabilitation program’ (BRP) group and a control (C) group. The BRP group received bedside rehabilitation schedules exercise programs by ward nurses. while the C group did not. Both groups received routine rehabilitation schedule. The Functional Independence Measure was used to measure outcome at the first, 14th and the 28th day of rehabilitation ward admitted. Data were analyzed using SPSS 22.0. After implementation of standardized ‘‘bedside rehabilitation program’, the results were: (1) the increasing of bedside rehabilitation had significant difference (p<.05) in promotion ADL function of stroke patients (2) the extend time of the bedside rehabilitation has significant difference (p<.05) in promotion ADL function of stroke patients compared with the control group. This study demonstrated that the ‘bedside rehabilitation program’ enhanced the ADL function in stroke patients. The nurses and rehabilitation ward managers need to understand that the extend time and frequency of rehabilitation provide a chance to enhanced the ADL function of stroke patients.

Keywords: stroke, bedside rehabilitation, functional activity, ADL

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Authors: El Kolli Meriem, Laouer Hocine, Sahli Farida, Akkal Salah, El Kolli Hayet

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The aerial parts of Daucus gracilis (Apiaceae) were subjected to hydrodistillation by a Clevenger apparatus to obtain the essential oil (EO) which has been analyzed by Gas Chromatography (GC) and GC coupled with mass spectrometry. The antioxidant properties of this EO and D. gracilis methanolic extract were studied by both of the free diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and the reducing power techniques. The dominant constituents of the EO were the elemicin (35.3 %) and the geranyl acetate (26.8 %). Both of EO and methanolic extract showed important antioxidant properties with respectively IC50 of 0,002 mg/ml and 0.06 mg/ml. They showed also a reducing power dose-dependent.

Keywords: daucus gracilis, apiaceae, essential oil, antioxidant activity

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Authors: Wu Chao

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The urban design of the new district in China is different from other existing cities at the city planning level, including Beijing, Shanghai, Guangzhou, etc. And the urban problems of these super-cities are same as many big cities around the world. The goal of the new district construction plan is to enable people to live comfortably, to improve the well-being of residents, and to create a way of life different from that of other urban communities. To avoid the emergence of the super community, the idea of "decentralization" is taken as the overall planning idea, and the function and form of each community are set up with a homogeneous allocation of resources so that the community can grow naturally. Similar to the growth of vines in nature, each community groups are independent and connected through roads, with clear community boundaries that limit their unlimited expansion. With a community contained 20,000 people as a case, the community is a mixture for living, production, office, entertainment, and other functions. Based on the development of the Internet, to create more space for public use, and can use data to allocate resources in real time. And this kind of shared space is the main part of the activity space in the community. At the same time, the transformation of spatial function can be determined by the usage feedback of all kinds of existing space, and the use of space can be changed by the changing data. Take the residential unit as the basic building function mass, take the lower three to four floors of the building as the main flexible space for use, distribute functions such as entertainment, service, office, etc. For the upper living space, set up a small amount of indoor and outdoor activity space, also used as shared space. The transformable space of the bottom layer is evenly distributed, combined with the walking space connected the community, the service and entertainment network can be formed in the whole community, and can be used in most of the community space. With the basic residential unit as the replicable module, the design of the other residential units runs through the idea of decentralization and the concept of the vine community, and the various units are reasonably combined. At the same time, a small number of office buildings are added to meet the special office needs. The new functional mixed community can change many problems of the present city in the future construction, at the same time, it can keep its vitality through the adjustment function of the Internet.

Keywords: decentralization, mixed functional community, shared space, spatial usage data

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Authors: Radostina Popova

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The paper presents an analysis of the innovation performance of small and medium-sized furniture enterprises in Bulgaria, accounting for over 97% of the companies in the sector. It contains advanced features of innovation in enterprises, specific features of the furniture industry in Bulgaria and analysis of the results of studies on the topic. The results from studies of three successive periods - 2006-2008; 2008-2010; 2010-2012, during which were studied 594 small and medium-sized furniture enterprises. There are commonly used in the EU definitions and indicators (European Commission, OECD, Oslo Manual), which allows for the comparability of results.

Keywords: innovation activity, competitiveness of innovation, furniture enterprises in Bulgaria

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Authors: Venere Ferraro, Silvia Ferraris

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We live in a world pervaded by new advanced technologies that have been changing the way we live and experience the surrounded. Besides, new technologies enable product innovation at different levels. Nevertheless, innovation does not lie just in the technological development and in its hard aspects but also in the meaningful use of it for the final user. In order to generate innovative products, a new perspective is needed: The shift from an instrument-oriented view of the technology towards a broader view that includes aspects like aesthetics, acceptance, comfort, and sociability. In many businesses, the user experience of the product is considered the key battlefield to achieve product innovation. (Holland 2011) The use of new technologies is indeed useless without paying attention to the user experience. This paper presents a workshop activity conducted at Design School of Politecnico di Milano in collaboration with Chiba University and aimed at generating innovative design concepts of high-tech mask. The students were asked to design the user experience of a new mask by exploiting emerging technologies such as wearable sensors and information communication technology (ICT) for a chosen field of application: safety or sport. When it comes to the user experience, the mask is a very challenging design product, because it covers aspects of product interaction and, most important, psychological and cultural aspects related to the impact on the facial expression. Furthermore, since the mask affects the face expression quite a lot, it could be a barrier to hide with, or it could be a mean to enhance user’s communication to others. The main request for the students was to take on a user-centered approach: To go beyond the instrumental aspects of product use and usability and focus on the user experience by shaping the technology in a desirable and meaningful way for the user reasoning on the metaphorical and cultural level of the product. During the one-week workshop students were asked to face the design process through (i) the research phase: an in-deep analysis of the user and field of application (safety or sport) to set design spaces (brief) and user scenario; (ii) the idea generation, (iii) the idea development. This text will shortly go through the meaning of the product innovation, the use and application of wearable technologies and will then focus on the user experience design in contrast with the technology-driven approach in the field of product innovation. Finally authors will describe the workshop activity and the concepts developed by the students stressing the important role of the user experience design in new product development.

Keywords: product innovation, user experience, technological exploitation, wearable technologies

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Authors: M. Maciejewska, F. Walkiewicz

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The aim of this work was to study the activity of several novel benzalkonium and alkylammonium and alkylimidazolium ionic liquids with 2-mercaptobenzothiazolate for use as accelerators in the sulphur vulcanisation of butadiene-styrene elastomer (SBR). The application of novel ionic liquids allowed for the elimination of N-cyclohexyl-2-benzothiazolesulfenamide from SBR compounds and for the considerable reduction of the amount of 2-mercaptobenzothiazole present in rubber products, which is favourable because, it is an allergenic agent. Synthesised salts could be used alternatively to standard accelerators in the vulcanisation of SBR, without any detrimental effects on the vulcanisation process, the physical properties or the thermal stability of the obtained vulcanisates. Ionic liquids increased the crosslink density of the vulcanisates and improved their thermal stability.

Keywords: ionic liquids, mechanical properties, styrene-butadiene rubber, vulcanisation

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Authors: Ö. Şenyiğit, A. Çolak

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It is of great importance for an architectural student to know the parameters through which he/she can conduct his/her design and makes his/her design effective in architectural education. Therefore; an empirical application study was carried out through the designing activity using the emulation model to support the design and design approaches of architectural students. During the investigation period, studies were done on the basic design elements and principles of the fall semester, and the emulation model, one of the designing methods that constitute the subject of the study, was fictionalized as three phased “recognition-interpretation-application”. As a result of the study, it was observed that when students were given a key method during the design process, their awareness increased and their aspects improved as well.

Keywords: basic design, design education, design methods, emulation

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Authors: Matthew Ocheleka Itodo, Ogo Agbo Ogo, Agnes Ogbene Abutu, Bawa Inalegwu

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Hyperlipidemia is characterised by elevated serum total cholesterol and low density and very low-density lipoprotein cholesterol and decreased high-density lipoprotein are the risk factor for coronary heart diseases. There are claims by traditional medicine practitioners in Nigeria that Moringa oleifera plants are used for the treatment of cardiovascular diseases, but it appears there is no scientific research and, publication or documented work to verify these claims. This study aimed to determine the effect of methanol root extracts of Moringa oleifera on Lipid profile, Atherogenic indices and 3 hydroxyl 3 methylglutaryl Coenzyme A reductase activity of poloxamer 407-induced hyperlipidemic rats. The animals were grouped into 8; Group 1: Normal control, Group 2: Hyperlipidemic control. Groups 2 to 8 were induced with Poloxamer 407 1000 mg/Kg body weight. However, group 3 were treated with standard drugs (atorvastatin). Group 4 was treated with crude extract, and groups 5 to 8 were treated with purified fractions from column chromatography. The preliminary antihyperlipidemic study showed Methanol root extract at 200 mg/kg body weight significantly (p≤0.05) decreased total cholesterol, low-density lipoprotein, triacylglyceride, 3 hydroxyls 3 methylglutaryl Coenzyme A reductase, and increase high-density lipoprotein of hyperlipidemic treated groups. Screening the extracts for the most potent anti-hyperlipidemic activity reveals that fraction 1 of Total Cholesterol and Fraction 3 of Triacylglyceride have the highest percentage reduction of 56% and 51%, respectively. The atherogenic risk factor of all induced treated rats shows a significant (p<0.05) decrease in levels of Castelli’s risk index II, atherogenic index of plasma and a significant (p<0.05) higher level of Castelli’s risk index I ratio. The study shows that the methanol extract of root possesses antihyperlipidemic effects and may explain why it has been found to be useful in the management of cardiovascular diseases by traditional medicine practitioners.

Keywords: hyperlipidemia, moringa oleifera, poloxamer 407, lipid profile

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Authors: Nilda Tri Putri, Muhammad Taufik

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Tools and agricultural machinery (Alsintan) is a tool used in agribusiness activities. Alsintan used to change the traditional farming systems generally use manual equipment into modern agriculture with mechanization. CV Nugraha Chakti Consultant make an action plan for industrial development Alsintan West Sumatra in 2012 to develop medium industries of Alsintan become a major industry of Alsintan, one of efforts made is increase the production capacity of the industry Alsintan. Production capacity for superior products as hydrotiller and threshers set each for 2.000 units per year. CV Citra Dragon as one of the medium industry alsintan in West Sumatra has a plan to relocate the existing plant to meet growing consumer demand each year. Increased production capacity and plant relocation plan has led to a change in the layout; therefore need to design the layout of the plant facility CV Citra Dragon. First step the to design of plant layout is design the layout of the production floor. The design of the production floor layout is done by applying group technology layout. The initial step is to do a machine grouping and part family using the Average Linkage Clustering (ALC) and Rank Order Clustering (ROC). Furthermore done independent work station design and layout design using the Modified Spanning Tree (MST). Alternative selection layout is done to select the best production floor layout between ALC and ROC cell grouping. Furthermore, to design the layout of warehouses, offices and other production support facilities. Activity Relationship Chart methods used to organize the placement of factory facilities has been designed. After structuring plan facilities, calculated cost manufacturing facility plant establishment. Type of layout is used on the production floor layout technology group. The production floor is composed of four cell machinery, assembly area and painting area. The total distance of the displacement of material in a single production amounted to 1120.16 m which means need 18,7minutes of transportation time for one time production. Alsintan Factory has designed a circular flow pattern with 11 facilities. The facilities were designed consisting of 10 rooms and 1 parking space. The measure of factory building is 84 m x 52 m.

Keywords: Average Linkage Clustering (ALC), Rank Order Clustering (ROC), Modified Spanning Tree (MST), Activity Relationship Chart (ARC)

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Authors: Tatsiana Ihnatovich, Darya Nizheharodava, Mikalai Halabarodzka, Tatsiana Savitskaya, Marina Zafranskaya

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Liver fibrosis is a complex of histological changes resulting from chronic liver disease accompanied by an excessive production and deposition of extracellular matrix components in the hepatic parenchyma. Liver fibrosis is a serious medical and social problem. Hepatic stellate cells (HSCs) make a significant contribution to the extracellular matrix deposition due to liver injury. Mesenchymal stem cells (MSCs) have a pronounced anti-inflammatory, regenerative and immunomodulatory effect; they are able to differentiate into hepatocytes and induce apoptosis of activated HSCs that opens the prospect of their use for preventing the excessive fibro-formation and the development of liver cirrhosis. The aim of the study is to evaluate the effect of MSCs therapy on the expression of fibrogenesis markers genes in liver tissue and HSCs cultures of rats with experimental liver cirrhosis (ELC). Materials and methods: ELC was induced by the common bile duct ligation (CBDL) in female Wistar rats (n = 19) with an average body weight of 250 (220 ÷ 270) g. Animals from the control group (n = 10) were sham-operated. On the 56th day after the CBDL, the rats of the experimental (n = 12) and the control (n = 5) groups received intraportal MSCs in concentration of 1×106 cells/animal (previously obtained from rat’s bone marrow) or saline, respectively. The animals were taken out of the experiment on the 21st day. HSCs were isolated by sequential liver perfusion in situ with following disaggregation, enzymatic treatment and centrifugation of cell suspension on a two-stage density gradient. The expression of collagen type I (Col1a1) and type III (Col3a1), matrix metalloproteinase type 2 (MMP2) and type 9 (MMP9), tissue inhibitor of matrix metalloproteinases type 1 (TIMP1), transforming growth factor β type 1 (TGFβ1) and type 3 (TGFβ3) was determined by real-time polymerase chain reaction. Statistical analysis was performed using Statistica 10.0. Results: In ELC rats compared to sham-operated animals, a significant increase of all studied markers expression was observed. The administration of MSCs led to a significant decrease of all detectable markers in the experimental group compared to rats without cell therapy. In ELC rats, an increased MMP9/TIMP1 ratio after cell therapy was also detected. The infusion of MSCs in the sham-operated animals did not lead to any changes. In the HSCs from ELC animals, the expression of Col1a1 and Col3a1 exceeded the similar parameters of the control group (p <0.05) and statistically decreased after the MSCs administration. The correlation between Col3a1 (Rs = 0.51, p <0.05), TGFβ1 (Rs = 0.6, p <0.01), and TGFβ3 (Rs = 0.75, p <0.001) expression in HSCs cultures and liver tissue has been found. Conclusion: Intraportal administration of MSCs to rats with ELC leads to a decreased Col1a1 and Col3a1, MMP2 and MMP9, TIMP1, TGFβ1 and TGFβ3 expression. The correlation between the expression of Col3a1, TGFβ1 and TGFβ3 in liver tissue and in HSCs cultures indicates the involvement of activated HSCs in the fibrogenesis that allows considering HSCs to be the main cell therapy target in ELC.

Keywords: cell therapy, experimental liver cirrhosis, hepatic stellate cells, mesenchymal stem cells

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Authors: Lamia Bouzgarrou, Irtyah Merchaoui, Amira Omrane, Salma Kammoun, Amine Daafa, Neila Chaari

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Backgrounds: Currently, low back pain (LBP) is one of the most prevalent public health problems, which caused severe morbidity among a large portion of the adult population. It is also associated with heavy direct and indirect costs, in particular, related to absenteeism and early retirement. Health care workers are one of most occupational groups concerned by LBP, especially because of biomechanical and psycho-organizational risk factors. Our current study aims to investigate risk factors associated with chronic low back pain among Tunisian caregivers in university-hospitals. Methods: Cross-sectional study conducted over a period of 14 months, with a representative sample of caregivers, matched according to age, sex and work department, in two university-hospitals in Tunisia. Data collection included items related to socio-professional characteristics, the evaluation of the working capacity index (WAI), the occupational stress (Karazek job strain questionnaire); the quality of life (SF12), the musculoskeletal disorders Nordic questionnaire, and the examination of the spine flexibility (distance finger-ground, sit-stand maneuver and equilibrium test). Results: Totally, 293 caregivers were included with a mean age equal to 42.64 ± 11.65 years. A body mass index (BMI) exceeding 30, was noted in 20.82% of cases. Moreover, no regular physical activity was practiced in 51.9% of cases. In contrast, domestic activity equal or exceeding 20 hours per week, was reported by 38.22%. Job strain was noted in 19.79 % of cases and the work capacity was 'low' to 'average' among 27.64% of subjects. During the 12 months previous to the investigation, 65% of caregivers complained of LBP, with pain rated as 'severe' or 'extremely severe' in 54.4% of cases and with a frequency of discomfort exceeding one episode per week in 58.52% of cases. During physical examination, the mean distance finger-ground was 7.10 ± 7.5cm. Caregivers assigned to 'high workload' services had the highest prevalence of LBP (77.4%) compared to other categories of hospital services, with no statistically significant relationship (P = 0.125). LBP prevalence was statistically correlated with female gender (p = 0.01) and impaired work capacity (p < 10⁻³). Moreover, the increase of the distance finger-ground was statistically associated with LBP (p = 0.05), advanced age (p < 10⁻³), professional seniority (p < 10⁻³) and the BMI ≥ 25 (p = 0.001). Furthermore, others physical tests of spine flexibility were underperformed among LBP suffering workers with a statistically significant difference (sit-stand maneuver (p = 0.03); equilibrium test (p = 0.01)). According to the multivariate analysis, only the domestic activity exceeding 20H/week, the degraded quality of physical life, and the presence of neck pain were significantly corelated to LBP. The final model explains 36.7% of the variability of this complaint. Conclusion: Our results highlighted the elevate prevalence of LBP among caregivers in Tunisian public hospital and identified both professional and individual predisposing factors. The preliminary analysis supports the necessity of a multidimensional approach to prevent this critical occupational and public health problem. The preventive strategy should be based both on the improvement of working conditions, and also on lifestyle modifications, and reinforcement of healthy behaviors in these active populations.

Keywords: health care workers, low back pain, prevention, risk factor

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Authors: Pierre-Louis Lucas, Corinne Loutelier-Bourhis, Narimane Mati-Baouche, Philippe Chan Tchi-Song, Patrice Lerouge, Elodie Mathieu-Rivet, Muriel Bardor

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N-glycosylation is a post-translational modification taking place in the Endoplasmic Reticulum and the Golgi apparatus where defined glycan features are added on protein in a very specific sequence Asn-X-Thr/Ser/Cys were X can be any amino acid except proline. Because it is well-established that those N-glycans play a critical role in protein biological activity, protein half-life and that a different N-glycan structure may induce an immune response, they are very important in Biopharmaceuticals which are mainly glycoproteins bearing N-glycans. From now, most of the biopharmaceuticals are produced by mammalian cells like Chinese Hamster Ovary cells (CHO) for their N-glycosylation similar to the human, but due to the high production costs, several other species are investigated as the possible alternative system. In this purpose, the green microalgae Chlamydomonas reinhardtii was investigated as the potential production system for Biopharmaceuticals. This choice was influenced by the facts that C. reinhardtii is a well-study microalgae which is growing fast with a lot of molecular biology tools available. This organism is also producing N-glycan on its endogenous proteins. However, the analysis of the N-glycan structure of this microalgae has revealed some differences as compared to the human. Rather than in Human where the glycans are processed by key enzymes called N-acetylglucosaminyltransferase I and II (GnTI and GnTII) adding GlcNAc residue to form a GlcNAc₂Man₃GlcNAc₂ core N-glycan, C. reinhardtii lacks those two enzymes and possess a GnTI independent glycosylation pathway. Moreover, some enzymes like xylosyltransferases and methyltransferases not present in human are supposed to act on the glycans of C. reinhardtii. Furthermore, the recent structural study by mass spectrometry shows that the N-glycosylation precursor supposed to be conserved in almost all eukaryotic cells results in a linear Man₅GlcNAc₂ rather than a branched one in C. reinhardtii. In this work, we will discuss the new released MS information upon C. reinhardtii N-glycan structure and their impact on our attempt to modify the glycan in a Human manner. Two strategies will be discussed. The first one consisted in the study of Xylosyltransferase insertional mutants from the CLIP library in order to remove xyloses from the N-glycans. The second will go further in the humanization by transforming the microalgae with the exogenous gene from Toxoplasma gondii having an activity similar to GnTI and GnTII with the aim to synthesize GlcNAc₂Man₃GlcNAc₂ in C. reinhardtii.

Keywords: Chlamydomonas reinhardtii, N-glycosylation, glycosyltransferase, mass spectrometry, humanization

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Authors: Iman Farasat, Howard M. Salis

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Engineered genetic circuits reprogram cellular behavior to act as living computers with applications in detecting cancer, creating self-controlling artificial tissues, and dynamically regulating metabolic pathways. Phenemenological models are often used to simulate and design genetic circuit behavior towards a desired behavior. While such models assume that each circuit component’s function is modular and independent, even small changes in a circuit (e.g. a new promoter, a change in transcription factor expression level, or even a new media) can have significant effects on the circuit’s function. Here, we use statistical thermodynamics to account for the several factors that control transcriptional regulation in bacteria, and experimentally demonstrate the model’s accuracy across 825 measurements in several genetic contexts and hosts. We then employ our first principles model to design, experimentally construct, and characterize a family of signal amplifying genetic circuits (genetic OpAmps) that expand the dynamic range of cell sensors. To develop these models, we needed a new approach to measuring the in vivo binding free energies of transcription factors (TFs), a key ingredient of statistical thermodynamic models of gene regulation. We developed a new high-throughput assay to measure RNA polymerase and TF binding free energies, requiring the construction and characterization of only a few constructs and data analysis (Figure 1A). We experimentally verified the assay on 6 TetR-homolog repressors and a CRISPR/dCas9 guide RNA. We found that our binding free energy measurements quantitatively explains why changing TF expression levels alters circuit function. Altogether, by combining these measurements with our biophysical model of translation (the RBS Calculator) as well as other measurements (Figure 1B), our model can account for changes in TF binding sites, TF expression levels, circuit copy number, host genome size, and host growth rate (Figure 1C). Model predictions correctly accounted for how these 8 factors control a promoter’s transcription rate (Figure 1D). Using the model, we developed a design framework for engineering multi-promoter genetic circuits that greatly reduces the number of degrees of freedom (8 factors per promoter) to a single dimensionless unit. We propose the Ptashne (Pt) number to encapsulate the 8 co-dependent factors that control transcriptional regulation into a single number. Therefore, a single number controls a promoter’s output rather than these 8 co-dependent factors, and designing a genetic circuit with N promoters requires specification of only N Pt numbers. We demonstrate how to design genetic circuits in Pt number space by constructing and characterizing 15 2-repressor OpAmp circuits that act as signal amplifiers when within an optimal Pt region. We experimentally show that OpAmp circuits using different TFs and TF expression levels will only amplify the dynamic range of input signals when their corresponding Pt numbers are within the optimal region. Thus, the use of the Pt number greatly simplifies the genetic circuit design, particularly important as circuits employ more TFs to perform increasingly complex functions.

Keywords: transcription factor, synthetic biology, genetic circuit, biophysical model, binding energy measurement

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Authors: Fereshteh Falah, Alireza Vasiee, Farideh Tabatabaei-Yazdi

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In this research, we investigated the usage of dairy sludge in the fermentation process and cyclosporin production. This bioactive compound is a metabolite produced by Tolypocladium inflatum. Results showed that about 200 ppm of cyclosporin can be produced in this fermentation. In order to have a proper and specific function, CyA must be free of any impurities, so we need purification. In this downstream processing, we used chromatographic extraction and evaluation of pharmacological activities of cyA. Results showed that the obtained metabolite has very high activity against Aspergilus niger (25mm clear zone). This cyclosporin was isolated for use as an antibiotic. The current research shows that this drug is very vital and commercially very important.

Keywords: fermentation, cyclosporin A, Tolypocladium inflatum, TLC

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Authors: Nataliya V. Dolgushina, Elena A. Gorodnova, Olga S. Beznoshenco, Andrey Yu Romanov, Irina V. Menzhinskaya, Lyubov V. Krechetova, Gennady T. Suchich

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In patients with COVID-19, generalized hypercoagulability can lead to the development of severe coagulopathy. This event is accompanied by the development of a pronounced inflammatory reaction. The observational prospective study included 39 patients with mild COVID-19 and 102 patients with moderate and severe COVID-19. Patients were then stratified into groups depending on the risk of venous thromboembolism. vWF to ADAMTS-13 concentrations and activity ratios were significantly higher in patients with a high venous thromboembolism risks in patients with moderate and severe forms COVID-19.

Keywords: ADAMTS-13, COVID-19, hypercoagulation, thrombosis, von Willebrand factor

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Authors: Yawen Dai, Ping Cheng, Jian Ru Gong

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Photoelctrochemical (PEC) water splitting using semiconductors (SCs) provides a convenient way to convert sustainable but intermittent solar energy into clean hydrogen energy, and it has been regarded as one of most promising technology to solve the energy crisis and environmental pollution in modern society. However, the record energy conversion efficiency of a PEC cell (~3%) is still far lower than the commercialization requirement (~10%). The sluggish kinetics of oxygen evolution reaction (OER) half reaction on photoanodes is a significant limiting factor of the PEC device efficiency, and electrocatalysts (ECs) are always deposited on SCs to accelerate the hole injection for OER. However, an active EC cannot guarantee enhanced PEC performance, since the newly emerged SC-EC interface complicates the interfacial charge behavior. Herein, α-Fe2O3 photoanodes coated with Co3O4 and CoO ECs are taken as the model system to glean fundamental understanding on the EC-dependent interfacial charge behavior. Intensity modulated photocurrent spectroscopy and electrochemical impedance spectroscopy were used to investigate the competition between interfacial charge transfer and recombination, which was found to be dominated by the charge storage capacities of ECs. The combined results indicate that both ECs can store holes and increase the hole density on photoanode surface. It is like a double-edged sword that benefit the multi-hole participated OER, as well as aggravate the SC-EC interfacial charge recombination due to the Coulomb attraction, thus leading to a nonmonotonic PEC performance variation trend with the increasing surface hole density. Co3O4 has low hole storage capacity which brings limited interfacial charge recombination, and thus the increased surface holes can be efficiently utilized for OER to generate enhanced photocurrent. In contrast, CoO has overlarge hole storage capacity that causes severe interfacial charge recombination, which hinders hole transfer to electrolyte for OER. Therefore, the PEC performance of α-Fe2O3 is improved by Co3O4 but decreased by CoO despite the similar electrocatalytic activity of the two ECs. First-principle calculation was conducted to further reveal how the charge storage capacity depends on the EC’s intrinsic property, demonstrating that the larger hole storage capacity of CoO than that of Co3O4 is determined by their Co valence states and original Fermi levels. This study raises up a new strategy to manipulate interfacial charge behavior and the resultant PEC performance by the charge storage capacity of ECs, providing insightful guidance for the interface design in PEC devices.

Keywords: charge storage capacity, electrocatalyst, interfacial charge behavior, photoelectrochemistry, water-splitting

Procedia PDF Downloads 142

Authors: Roya Bakhtiar, Alireza Abdolmohammadi, Hadi Hajarian, Zahra Nikousefat, Davood, Kalantar-Neyestanaki

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The objective of the present study was to determine the polymorphism in the leptin (332G>A) and its association with biometric traits in Sanjabi sheep. For this purpose, blood samples from 96 rams were taken, and tail length, width tail, circumference tail, body length, body width, and height were simultaneously recorded. PCR was performed using specific primer to amplify 463 bp fragment including exon 3 of leptin gene, and PCR products were digested by Cail restriction enzymes. The 332G>A (at 332th nucleotide of exon 3 leptin gene) that caused an amino acid change from Arg to Gln was detected by Cail (CAGNNNCTG) endonuclease, as the endonuclease cannot cut this region if G nucleotide is located in this position. Three genotypes including GG (463), GA (463, 360and 103 bp) and GG (360 bp and 103 bp) were identified after digestion by enzyme. The estimated frequencies of three genotypes including GG, GA, and AA for 332G>A locus were 0.68, 0.29 and 0.03 and those were 0.18 and 0.82 for A and G alleles, respectively. In the current study, chi-square test indicated that 332G>A positions did not deviate from the Hardy–Weinberg (HW) equilibrium. The most important reason to show HW equation was that samples used in this study belong to three large local herds with a traditional breeding system having random mating and without selection. Shannon index amount was calculated which represent an average genetic variation in Sanjabi rams. Also, heterozygosity estimated by Nei index indicated that genetic diversity of mutation in the leptin gene is moderate. Leptin gene polymorphism in the 332G>A had significant effect on body length (P<0.05) trait, and individuals with GA genotype had significantly the higher body length compared to other individuals. Although animals with GA genotype had higher body width, this difference was not statistically significant (P>0.05). This non-synonymous SNP resulted in different amino acid changes at codon positions111(R/Q). As leptin activity is localized, at least in part, in domains between amino acid residues 106-1406, it is speculated that the detected SNP at position 332 may affect the activity of leptin and may lead to different biological functions. Based to our results, due to significant effect of leptin gene polymorphism on body size traits, this gene may be used a candidate gene for improving these traits.

Keywords: body size, Leptin gene, PCR-RFLP, Sanjabi sheep

Procedia PDF Downloads 345

Authors: Omar Pillaca-Pullo, Arturo Alejandro-Paredes, Carol Flores-Fernandez, Marijuly Sayuri Kina, Amparo Iris Zavaleta

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Aqueous two-phase system (ATPS) is an interesting alternative for separating industrial enzymes due to it is easy to scale-up and low cost. Polyethylene glycol (PEG) mixed with potassium phosphate or magnesium sulfate is one of the most frequently polymer/salt ATPS used, but the consequences of its use is a high concentration of phosphates and sulfates in wastewater causing environmental issues. Citrate could replace these inorganic salts due to it is biodegradable and does not produce toxic compounds. On the other hand, statistical design of experiments is widely used for ATPS optimization and it allows to study the effects of the involved variables in the purification, and to estimate their significant effects on selected responses and interactions. The 24 factorial design with four central points (20 experiments) was employed to study the partition and purification of proteases produced by Pseudomonas sp. in PEG/citrate ATPS system. ATPS was prepared with different sodium citrate concentrations [14, 16 and 18% (w/w)], pH values (7, 8 and 9), PEG molecular weight (2,000; 4,000 and 6,000 g/mol) and PEG concentrations [18, 20 and 22 % (w/w)]. All system components were mixed with 15% (w/w) of the fermented broth and deionized water was added to a final weight of 12.5 g. Then, the systems were mixed and kept at room temperature until to reach two-phases separation. Volumes of the top and bottom phases were measured, and aliquots from both phases were collected for subsequent proteolytic activity and total protein determination. Influence of variables such as PEG molar mass (MPEG), PEG concentration (CPEG), citrate concentration (CSal) and pH were evaluated on the following responses: purification factor (PF), activity yield (Y), partition coefficient (K) and selectivity (S). STATISTICA program version 10 was used for the analysis. According to the obtained results, higher levels of CPEG and MPEG had a positive effect on extraction, while pH did not influence on the process. On the other hand, the CSal could be related with low values of Y because of the citrate ions have a negative effect on solubility and enzymatic structure. The optimum values of Y (66.4 %), PF (1.8), K (5.5) and S (4.3) were obtained at CSal (18%), MPEG (6,000 g/mol), CPEG (22%) and pH 9. These results indicated that the PEG/citrate system is accurate to purify these Pseudomonas sp. proteases from fermented broth as a first purification step.

Keywords: citrate, polyethylene glycol, protease, Pseudomonas sp

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Authors: Andrew Hall, Paul Clarkson

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Climate change is predicted to increase the number of extreme weather events intensifying the strain on Railway Earthworks. This paper describes the use of Distributed Rayleigh Sensing to monitor low frequency activity on a vulnerable earthworks sectionprone to landslides alongside a railway line in Northern Spain. The vulnerable slope is instrumented with conventional slope stability sensors allowing an assessment to be conducted of the application of Distributed Rayleigh Sensing as an earthwork condition monitoring tool to enhance the resilience of railway networks.

Keywords: condition monitoring, railway earthworks, distributed rayleigh sensing, climate change

Procedia PDF Downloads 209

Authors: Yudhitya Kusumawati, Yuki Mitsuya, Tomooki Shiba, Mitsuru Uesaka

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After the Fukushima Daiichi nuclear power reactor incident, there are a lot of unaccountable nuclear fuel debris in the reactor core area, which is subject to safeguard and criticality safety. Before the actual precise analysis is performed, preliminary on-site screening and mapping of nuclear debris activity need to be performed to provide a reliable data on the nuclear debris mass-extraction planning. Through a collaboration project with Japan Atomic Energy Agency, an on-site nuclear debris screening system by using dual energy X-Ray inspection and neutron energy resonance analysis has been established. By using the compact and mobile pulsed neutron source constructed from 3.95 MeV X-Band electron linac, coupled with Tungsten as electron-to-photon converter and Beryllium as a photon-to-neutron converter, short-distance neutron Time of Flight measurement can be performed. Experiment result shows this system can measure neutron energy spectrum up to 100 eV range with only 2.5 meters Time of Flightpath in regards to the X-Band accelerator’s short pulse. With this, on-site neutron Time of Flight measurement can be used to identify the nuclear debris isotope contents through Neutron Resonance Transmission Analysis (NRTA). Some preliminary NRTA experiments have been done with Tungsten sample as dummy nuclear debris material, which isotopes Tungsten-186 has close energy absorption value with Uranium-238 (15 eV). The results obtained shows that this system can detect energy absorption in the resonance neutron area within 1-100 eV. It can also detect multiple elements in a material at once with the experiment using a combined sample of Indium, Tantalum, and silver makes it feasible to identify debris containing mixed material. This compact neutron Time of Flight measurement system is a great complementary for dual energy X-Ray Computed Tomography (CT) method that can identify atomic number quantitatively but with 1-mm spatial resolution and high error bar. The combination of these two measurement methods will able to perform on-site nuclear debris screening at Fukushima Daiichi reactor core area, providing the data for nuclear debris activity mapping.

Keywords: neutron source, neutron resonance, nuclear debris, time of flight

Procedia PDF Downloads 239

Authors: Bechir Hbibi, Lamine Mili

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Understanding the condition of comatose patients can be difficult, but it is crucial to their optimal treatment. Consequently, numerous scoring systems have been developed around the world to categorize patient states based on physiological assessments. Although validated and widely adopted by medical communities, these scores still present numerous limitations and obstacles. Even with the addition of additional tests and extensions, these scoring systems have not been able to overcome certain limitations, and it appears unlikely that they will be able to do so in the future. On the other hand, physiological tests are not the only way to extract ideas about comatose patients. EEG signal analysis has helped extensively to understand the human brain and human consciousness and has been used by researchers in the classification of different levels of disease. The use of EEG in the ICU has become an urgent matter in several cases and has been recommended by medical organizations. In this field, the EEG is used to investigate epilepsy, dementia, brain injuries, and many other neurological disorders. It has recently also been used to detect pain activity in some regions of the brain, for the detection of stress levels, and to evaluate sleep quality. In our recent findings, our aim was to use multifractal analysis, a very successful method of handling multifractal signals and feature extraction, to establish a state of awareness scale for comatose patients based on their electrical brain activity. The results show that this score could be instantaneous and could overcome many limitations with which the physiological scales stock. On the contrary, multifractal analysis stands out as a highly effective tool for characterizing non-stationary and self-similar signals. It demonstrates strong performance in extracting the properties of fractal and multifractal data, including signals and images. As such, we leverage this method, along with other features derived from EEG signal recordings from comatose patients, to develop a scale. This scale aims to accurately depict the vigilance state of patients in intensive care units and to address many of the limitations inherent in physiological scales such as the Glasgow Coma Scale (GCS) and the FOUR score. The results of applying version V0 of this approach to 30 patients with known GCS showed that the EEG-based score similarly describes the states of vigilance but distinguishes between the states of 8 sedated patients where the GCS could not be applied. Therefore, our approach could show promising results with patients with disabilities, injected with painkillers, and other categories where physiological scores could not be applied.

Keywords: coma, vigilance state, EEG, multifractal analysis, feature extraction

Procedia PDF Downloads 78

Authors: S. T. Kumbhar, M. S. Bhatia, R. C. Khairate

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An effective nanodrug delivery system was developed by using chitosan for increased encapsulation efficiency and retarded release of curcumin. Potential ionotropic gelation method was used for the development of chitosan nanoparticles with TPP as cross-linker. The characterization was done for analysis of size, structure, surface morphology, and thermal behavior of synthesized chitosan nanoparticles. The encapsulation efficiency was more than 80%, with improved drug loading capacity. The in-vitro drug release study showed that curcumin release rate was decreased significantly. These chitosan nanoparticles could be a suitable platform for co-delivery of curcumin and anticancer agent for enhanced cytotoxic effect on tumor cells.

Keywords: Curcumin, chitosan, nanoparticles, anticancer activity

Procedia PDF Downloads 181

Authors: Zeinab Hmama

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Much recent discourse has highlighted the supporting role of social entrepreneurs in solving social problems. However, the identification of social enterprise’s characteristics in emerging countries has not yet been thoroughly examined. This research seeks to explores the profile of social enterprises in Morocco. In this perspective, we conduct a quantitative study on a sample of 87 social enterprises. This study was undertaken in Morocco based on a quantitative study lead among sample consists of 82 organizations qualified as social enterprises. Therefore, the response rate was 45.12% (37/82). Participants in the study were described on the basis of the following demographic characteristics: Gender, Age, Education Level and field, Entrepreneurial activity age, Legal forms, Line of business.

Keywords: social entrepreneurship, social enterprise, problem resolution, value creation

Procedia PDF Downloads 157

Authors: Ila Shukla, Lubna Azmi, Shyam Sunder Gupta, C. V. Rao

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Treatments against Hepatitis-C virus (HCV) are already available, but the current high cost of such treatments limit them to wealthy patients only. Hence our current study is aimed at the rectification of HCV infection by using Lichen rangiferinus (LRE) extract in in vitro cultures. Anti-HCV activity of the given extract was evaluated using the virus grown in cell culture (HCVcc). Two control inhibitors, erlotinib and telaprevir, were systematically included in each experiment. At the end of the incubation period, we evaluated cell viability and viral replication. The LRE inhibited the growth of HCV in a dose dependent manner.

Keywords: Erlotinib, Hepatitis C, Lichen rangiferinus, Telaprevir

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Authors: Olfa Kaabia, Ilyes Abid, Khaled Guesmi

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This pioneer paper studies whether and how Bitcoin shocks are transmitted to the U.S economy. We employ a new methodology: TVP FAVAR model with stochastic volatility. We use a large dataset of 111 major U.S variables from 1959:m1 to 2016:m12. The results show that Bitcoin shocks significantly impact the U.S. economy. This significant impact is pronounced in a volatile and increasing U.S economy. The Bitcoin has a positive relationship on the U.S real activity, and a negative one on U.S prices and interest rates. Effects on the Monetary Policy exist via the inter-est rates and the Money, Credit and Finance transmission channels.

Keywords: bitcoin, US economy, FAVAR models, stochastic volatility

Procedia PDF Downloads 249

Authors: Fongang Robert Tichuck

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This paper reports the findings of a study undertaken to assess the socio-economic and environmental impacts of mining in Bertoua Eastern Region of Cameroon. In addition to sampling community perceptions of mining activities, the study prescribes interventions that can assist in mitigating the negative impacts of mining. Marked environmental and interrelated socio-economic improvements can be achieved within regional artisanal gold mines if the government provides technical support to local operators, regulations are improved, and illegal mining activity is reduced.

Keywords: gold mining, socio-economic, mining activities, local people

Procedia PDF Downloads 399