Search results for: buffer overflow

Authors: Jennifer A. Kowalkowski

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Production agriculture (farming) is a physically, emotionally, and cognitively stressful occupation, where workers have little control over the stressors that impact both their work and their lives. In an occupation already rife with hazards, these occupational-related stressors have been shown to increase farm workers’ risks for illness, injury, disability, and death associated with their work. Despite efforts to mitigate the negative health effects from occupational-related stress (ORS) and to promote health and well-being (HWB) among farmers in the US, marked improvements have not been attained. Social support accessed through social networks has been shown to buffer against the negative health effects from stress, yet no studies have directly examined these relationships among farmers. The purpose of this study was to use social network analysis to explore the social networks of farm owner-operators and the social supports available to them for mitigating the negative health effects of ORS. A convenience sample of 71 farm owner-operators from a Midwestern County in the US completed and returned a mailed survey (55.5% response rate) that solicited information about their social networks related to ORS. Farmers reported an average of 2.4 individuals in their personal networks and higher levels of comfort discussing ORS with female network members. Farmers also identified few connections (3.4% density) and indicated low comfort with members of affiliation networks specific to ORS. Findings from this study highlighted that farmers accessed different social networks and resources for their personal HWB than for issues related to occupational(farm-related) health and safety. In addition, farmers’ social networks for personal HWB were smaller, with different relational characteristics than reported in studies of farmers’ social networks related to occupational health and safety. Collectively, these findings suggest that farmers conceptualize personal HWB differently than farm health and safety. Therefore, the same research approaches and targets that guide occupational health and safety research may not be appropriate for personal HWB for farmers. Interventions and programming targeting ORS and HWB have largely been offered through the same platforms or mechanisms as occupational health and safety programs. This may be attributed to the significant overlap between the farm as a family business and place of residence, or that ORS stems from farm-related issues. However, these assumptions translated to health research of farmers and farm families from the occupational health and safety literature have not been directly studied or challenged. Thismay explain why past interventions have not been effective at improving health outcomes for farmers and farm families. A close examination of findings from this study raises important questions for researchers who study agricultural health. Findings from this study have significant implications for future research agendas focused on addressing ORS, HWB, and health disparities for farmersand farm families.

Keywords: agricultural health, occupational-related stress, social networks, well-being

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Authors: Zaima Umar, Anas S. Qureshi, Adeel Sarfraz, Saqib Umar, Talha Umar, Muhammad Usman

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Ostrich has been a good source of food and income for people across the world. To get a better understanding of health and health-related problems, the knowledge of its digestive system is of utmost importance. The present study was conducted to determine the morphological and histometrical variations in the digestive system and associated glands of ostrich (Struthio camelus) as regard to the gender and progressive age. A total of 40 apparently healthy ostriches of both genders and two progressive age groups; young one (less than two year, group A); and adult (2-15 years, group B) in equal number were used in this study. Digestive organs including tongue, esophagus, proventriculus, gizzard, small and large intestines and associated glands like liver and pancreas were collected immediately after slaughtering the birds. The organs of the digestive system and associated glands of each group were studied grossly and histologically. Grossly colour, shape consistency, weight and various dimensions (length, width, and circumference) of organs of the digestive tract and associated glands were recorded. The mean (± SEM) of all gross anatomical parameters in group A were significantly (p ≤ 0.01) different from that of group B. For microscopic studies, 1-2 cm tissue samples of organs of the digestive system and associated glands were taken. The tissue was marked and fixed in the neutral buffer formaldehyde solution for histological studies. After fixation, the sections of 5-7 µm were cut and stained by haematoxylin and eosin stain. All the layers (epithelium, lamina propria, lamina muscularis, submucosa and tunica muscularis) were measured (µm) with the help of automated computer software Image J®. The results of this study provide valuable information on the gender and age-related histological and histometrical variations in the digestive organs of ostrich (Struthio camelus). The microscopic studies of different parts of the digestive system revealed highly significant differences (p ≤ 0.01) among the two groups. The esophagus was lined by non-keratinized stratified squamous epithelium. The duodenum, jejunum, and ileum showed similar histological structures. Statistical analysis revealed significant (p ≤ 0.05) increase in the thickness of different tunics of the gastrointestinal tract in adult birds (up to 15 years) as compared with young ones (less than two years). Therefore, it can be concluded that there is a gradual but consistent growth in the observed digestive organs mimicking that of other poultry species and may be helpful in determining the growth pattern in this bird. However, there is a need to record the changes at closer time intervals.

Keywords: ostrich, digestive system, histomorphometry, grossly

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Authors: Ciwang Teyra, A. H. Y. Lai

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Introduction: Indigenous LGBTQ individuals face with significant life stress such as racial and gender discrimination and microaggressions, which may lead to negative impacts of their mental health. Although studies relevant to Taiwanese indigenous LGBTQpeople gradually increase, most of them are primarily conceptual or qualitative in nature. This research aims to fulfill the gap by offering empirical quantitative evidence, especially investigating the impact of racial and gender microaggressions on mental health among Taiwanese indigenous LGBTQindividuals with an intersectional perspective, as well as examine whether social support can help them to cope with microaggressions. Methods: Participants were (n=200; mean age=29.51; Female=31%, Male=61%, Others=8%). A cross-sectional quantitative design was implemented using data collected in the year 2020. Standardised measurements was used, including Racial Microaggression Scale (10 items), Gender Microaggression Scale (9 items), Social Support Questionnaire-SF(6 items); Patient Health Questionnaire(9-item); and Generalised Anxiety Disorder(7-item). Covariates were age, gender, and perceived economic hardships. Structural equation modelling (SEM) was employed using Mplus 8.0 with the latent variables of depression and anxiety as outcomes. A main effect SEM model was first established (Model1).To test the moderation effects of perceived social support, an interaction effect model (Model 2) was created with interaction terms entered into Model1. Numerical integration was used with maximum likelihood estimation to estimate the interaction model. Results: Model fit statistics of the Model 1:X2(df)=1308.1 (795), p<.05; CFI/TLI=0.92/0.91; RMSEA=0.06; SRMR=0.06. For Model, the AIC and BIC values of Model 2 improved slightly compared to Model 1(AIC =15631 (Model1) vs. 15629 (Model2); BIC=16098 (Model1) vs. 16103 (Model2)). Model 2 was adopted as the final model. In main effect model 1, racialmicroaggressionand perceived social support were associated with depression and anxiety, but not sexual orientation microaggression(Indigenous microaggression: b = 0.27 for depression; b=0.38 for anxiety; Social support: b=-0.37 for depression; b=-0.34 for anxiety). Thus, an interaction term between social support and indigenous microaggression was added in Model 2. In the final Model 2, indigenous microaggression and perceived social support continues to be statistically significant predictors of both depression and anxiety. Social support moderated the effect of indigenous microaggression of depression (b=-0.22), but not anxiety. All covariates were not statistically significant. Implications: Results indicated that racial microaggressions have a significant impact on indigenous LGBTQ people’s mental health. Social support plays as a crucial role to buffer the negative impact of racial microaggression. To promote indigenous LGBTQ people’s wellbeing, it is important to consider how to support them to develop social support network systems.

Keywords: microaggressions, intersectionality, indigenous population, mental health, social support

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Authors: Rajeev Ravindran, Amit K. Jaiswal

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Lignocellulose is the largest reservoir of inexpensive, renewable source of carbon. It is composed of lignin, cellulose and hemicellulose. Cellulose and hemicellulose is composed of reducing sugars glucose, xylose and several other monosaccharides which can be metabolised by microorganisms to produce several value added products such as biofuels, enzymes, aminoacids etc. Enzymatic treatment of lignocellulose leads to the release of monosaccharides such as glucose and xylose. However, factors such as the presence of lignin, crystalline cellulose, acetyl groups, pectin etc. contributes to recalcitrance restricting the effective enzymatic hydrolysis of cellulose and hemicellulose. In order to overcome these problems, pre-treatment of lignocellulose is generally carried out which essentially facilitate better degradation of lignocellulose. A range of pre-treatment strategy is commonly employed based on its mode of action viz. physical, chemical, biological and physico-chemical. However, existing pretreatment strategies result in lower sugar yield and formation of inhibitory compounds. In order to overcome these problems, we proposes a novel pre-treatment, which utilises the superior oxidising capacity of alkaline potassium permanganate assisted by ultra-sonication to break the covalent bonds in spent coffee waste to remove recalcitrant compounds such as lignin. The pre-treatment was conducted for 30 minutes using 2% (w/v) potassium permanganate at room temperature with solid to liquid ratio of 1:10. The pre-treated spent coffee waste (SCW) was subjected to enzymatic hydrolysis using enzymes cellulase and hemicellulase. Shake flask experiments were conducted with a working volume of 50mL buffer containing 1% substrate. The results showed that the novel pre-treatment strategy yielded 7 g/L of reducing sugar as compared to 3.71 g/L obtained from biomass that had undergone dilute acid hydrolysis after 24 hours. From the results obtained it is fairly certain that ultrasonication assists the oxidation of recalcitrant components in lignocellulose by potassium permanganate. Enzyme hydrolysis studies suggest that ultrasound assisted alkaline potassium permanganate pre-treatment is far superior over treatment by dilute acid. Furthermore, SEM, XRD and FTIR were carried out to analyse the effect of the new pre-treatment strategy on structure and crystallinity of pre-treated spent coffee wastes. This novel one-step pre-treatment strategy was implemented under mild conditions and exhibited high efficiency in the enzymatic hydrolysis of spent coffee waste. Further study and scale up is in progress in order to realise future industrial applications.

Keywords: spent coffee waste, alkaline potassium permanganate, ultra-sonication, physical characterisation

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Authors: Maico R. Severino, Laura G. Caixeta, Nadine M. Costa, Raísa L. T. Napoleão, Éverton F. V. Valle, Diego D. Calixto, Danielle Oliveira

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One of the biggest barriers that companies find nowadays is the coordination of production flow in their Supply Chains (SC). In this study, coordination is understood as a mechanism for incorporating the entire production channel, with everyone involved focused on achieving the same goals. Sometimes, this coordination is attempted by the use of logistics practices or production plan and control methods. No papers were found in the literature that presented the combined use of logistics practices and production plan and control methods. The main objective of this paper is to propose solutions for six case studies combining logistics practices and Ordering Systems (OS). The methodology used in this study was a conceptual model of decision making. This model contains six phases: a) the analysis the types and characteristics of relationships in the SC; b) the choice of the OS; c) the choice of the logistics practices; d) the development of alternative proposals of combined use; e) the analysis of the consistency of the chosen alternative; f) the qualitative and quantitative assessment of the impact on the coordination of the production flow and the verification of applicability of the proposal in the real case. This study was conducted on six Brazilian SC of different sectors: footwear, food and beverages, garment, sugarcane, mineral and metal mechanical. The results from this study showed that there was improvement in the coordination of the production flow through the following proposals: a) for the footwear industry the use of Period Bath Control (PBC), Quick Response (QR) and Enterprise Resource Planning (ERP); b) for the food and beverage sector firstly the use of Electronic Data Interchange (EDI), ERP, Continuous Replenishment (CR) and Drum-Buffer-Rope Order (DBR) (for situations in which the plants of both companies are distant), and secondly EDI, ERP, Milk-Run and Review System Continues (for situations in which the plants of both companies are close); c) for the garment industry the use of Collaborative Planning, Forecasting, and Replenishment (CPFR) and Constant Work-In-Process (CONWIP) System; d) for the sugarcane sector the use of EDI, ERP and CONWIP System; e) for the mineral processes industry the use of Vendor Managed Inventory (VMI), EDI and MaxMin Control System; f) for the metal mechanical sector the use of CONWIP System and Continuous Replenishment (CR). It should be emphasized that the proposals are exclusively recommended for the relationship between client and supplier studied. Therefore, it cannot be generalized to other cases. However, what can be generalized is the methodology used to choose the best practices for each case. Based on the study, it can be concluded that the combined use of OS and logistics practices enable a better coordination of flow production on SC.

Keywords: supply chain management, production flow coordination, logistics practices, ordering systems

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Authors: Izuddin Fahmy Abu, Mohamad Haiqal Nizar Mohamad, Muhammad Fattah Fazel, Renu Agarwal, Igor Iezhitsa, Nor Salmah Bakar, Henrik Franzyk, Ian Mellor

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Glaucoma is the global leading cause of irreversible blindness. Currently, the available treatment strategy only involves lowering intraocular pressure (IOP); however, the condition often progresses despite lowered or normal IOP in some patients. N-methyl-D-aspartate receptor (NMDAR) excitotoxicity often occurs in neurodegeneration-related glaucoma; thus it is a relevant target to develop a therapy based on neuroprotection approach. This study investigated the effects of Philanthotoxin-343 (PhTX-343), an NMDAR antagonist, on the neuroprotection of NMDA-induced glaucoma to alleviate visual impairments. Male Sprague-Dawley rats were equally divided: Groups 1 (control) and 2 (glaucoma) were intravitreally injected with phosphate buffer saline (PBS) and NMDA (160nM), respectively, while group 3 was pre-treated with PhTX-343 (160nM) 24 hours prior to NMDA injection. Seven days post-treatments, rats were subjected to visual behavior assessments and subsequently euthanized to harvest their retina and optic nerve tissues for histological analysis and determination of nitrosative stress level using 3-nitrotyrosine ELISA. Visual behavior assessments via open field, object, and color recognition tests demonstrated poor visual performance in glaucoma rats indicated by high exploratory behavior. PhTX-343 pre-treatment appeared to preserve visual abilities as all test results were significantly improved (p < 0.05). H&E staining of the retina showed a marked reduction of ganglion cell layer thickness in the glaucoma group; in contrast, PhTX-343 significantly increased the number by 1.28-folds (p < 0.05). PhTX-343 also increased the number of cell nuclei/100μm2 within inner retina by 1.82-folds compared to the glaucoma group (p < 0.05). Toluidine blue staining of optic nerve tissues showed that PhTX-343 reduced the degeneration changes compared to the glaucoma group which exhibited vacuolation overall sections. PhTX-343 also decreased retinal 3- nitrotyrosine concentration by 1.74-folds compared to the glaucoma group (p < 0.05). All results in PhTX-343 group were comparable to control (p > 0.05). We conclude that PhTX-343 protects against NMDA-induced changes and visual impairments in the rat model by reducing nitrosative stress levels.

Keywords: excitotoxicity, glaucoma, nitrosative stress , NMDA receptor , N-methyl-D-aspartate , philanthotoxin, visual behaviour

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Authors: Madelyn N. Moawad, Hermine R. Z. Tadros, Mary G. Ghobrial, Ahmad R. Bassiouny, Kamal M. Kandeel, Athar Ata

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Seaweeds are potential renewable resources in marine environment. They provide an excellent source of bioactive substances such as dietary fibers and various functional polysaccharides that could potentially be used as ingredients for both human and animal health applications. The observations suggested that these bioactive compounds have strong antioxidant properties, which have beneficial effects on human health. The present research aimed at finding new chemical products from local marine macroalgae for natural medicinal uses and consumption for their nutritional values. Macroalgae samples were collected manually mainly from the Mediterranean Sea at shallow subtidal zone of Abu Qir Bay, Alexandria, Egypt. The chemical compositions of lyophilized materials of six selected macroalgal species; Colpomenia sinuosa, Sargassum linifolium, Padina pavonia, Pterocladiella capillacea, Laurencia pinnatifidia, and Caulerpa racemosa, were investigated for proteins using bovine serum albumin, and carbohydrates were assayed by phenol-sulfuric acid reaction. The macroalgae lipid was extracted with chloroform, methanol and phosphate buffer. Vitamins were extracted using trichloroacetic acid. Chlorophylls and total carotenoids were determined spectrophotometrically and total phenols were extracted with methanol. In addition, lipid-soluble, and water-soluble antioxidant, and anti α-glucosidase activities were measured spectrophotometrically. The antioxidant activity of hexane extracts was investigated using phosphomolybdenum reagent. The anti-α-glucosidase effect measurement was initiated by mixing α-glucosidase solution with p-nitrophenyl α-D-glucopyranoside. The results showed that the ash contents varied from 11.2 to 35.4 % on dry weight basis for P. capillacea and Laurencia pinnatifidia, respectively. The protein contents ranged from 5.63 % in brown macroalgae C. sinuosa to 8.73 % in P. pavonia. A relative wide range in carbohydrate contents was observed (20.06–46.75 %) for the test algal species. The highest lipid percentage was found in green alga C. racemosa (5.91%) followed by brown algae P. pavonia (3.57%) and C. sinuosa (2.64%). The phenolic contents varied from 1.32 mg GAE/g for C. sinuosa to 4.00 mg GAE/g in P. pavonia. The lipid-soluble compounds exhibited higher antioxidant capacity (73.18-145.95 µM/g) than that of the water-soluble ones ranging from 24.83 µM/g in C. racemosa to 74.07 µM/g in S. linifolium. The most potent anti-α-glucosidase activity was observed for P. pavonia with IC50 of 17.12 μg/ml followed by S. linifolium (IC50 = 71.75 μg/ml), C. racemosa (IC50 = 84.73 μg/ml), P. capillacea (IC50 = 92.16 μg/ml), C. sinuosa (IC50 = 112.44 μg/ml), and L. pinnatifida (IC50 = 115.11 μg/ml).

Keywords: α-glucosidase, lyophilized, macroalgae, spectrophotometrically

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Authors: Dace Grauda, Gunta Cekstere, Inta Belogrudova, Andis Karlsons, Isaak Rashal

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Trifolium repens L. (white or Dutch clover) is a perennial herb, belongs to legume family (Leguminosae Juss.), spread extensively by stolons and seeds. The species is cultivated worldwide and was naturalized in many countries in meadows, yards, gardens, along roads and streets etc., especially in temperate regions. It is widespread also in grasslands throughout Riga, the capital of Latvia. The goal of this study was to investigate genetic structure of white clover population in Riga and to evaluate influence of different salt concentration on plants. For this purpose universal retrotranspozone based IRAP (Inter-Retrotransposon Amplified Polymorphism) method was used. The plant material was collected in different regions of Riga and in several urban areas of Latvia. Plant DNA was isolated from in silicogel dried leaves of using 1% CTAB (cetyltrimet-ammonium bromide) buffer DNA extraction procedure. Genetic structure of city population and wild populations were compared. Soil salinization is an important issue associated with low water resources and highly urbanized areas in aride and semi-aride climate conditions, as well as de-icing salt application to prevent ice formation on roads in winter. The T. repens variety ‘Daile’ (form giganteum), one of the often used component of urban greeneries, was studied in this investigation. Plants were grown from seeds and cultivated in the light conditions (18-25 C, 16h/8h of day/night, light intensity 3000 lx) in plastic pots (200 ml), filled with commercial neutralized (pH 5.9 ± 0.3) peat substrate with mineral nutrients. To analyse the impact of increased soil salinity treatments with gradually rising NaCl (0; 20; 40; 60; 80; 100 mM) levels were arranged. Plants were watered when necessary with deionised water to provide optimum substrate moisture 60-70%. The experiment was terminated six weeks after establishment. For analysis of mineral nutrients, dry plant material (above ground part and roots) was used. Decrease of Na content can be significant under elevated salinity till 20 mM NaCl. High NaCl concentrations in the substrate increase Na, Cl, Cu, Fe, and Mn accumulation, but reduce S, Mg, K content in the plant above ground parts. Abiotic stresses generally changes the levels of DNA metilation. Several candidate gene for salt tolerance will be analysed for DNA metilation level using Pyromark-Q24 advanced.

Keywords: DNA metilation, IRAP, soil salinization, white clover

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Authors: B. T. Naveen Kumar, Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, A. H. Shanthanagouda, Orawan Boodde, K. M. Shankar, Prakash Patil, Shubhkaramjeet Kaur

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Early mortality syndrome (EMS)/Acute Hepatopancreatic Necrosis Disease (AHPND) has emerged as a major obstacle for the shrimp farming around the world. It is caused by a strain of Vibrio parahaemolyticus. The possible preventive and control measure is, early and rapid detection of the pathogen in the broodstock, post-larvae and monitoring the shrimp during the culture period. Polymerase chain reaction (PCR) based early detection methods are good, but they are costly, time taking and requires a sophisticated laboratory. The present study was conducted to develop a simple, sensitive and rapid diagnostic farmer level kit for the reliable detection of AHPND in shrimp. A panel of monoclonal antibodies (MAbs) were raised against the recombinant Pir B protein (rPirB). First, an immunodot was developed by using MAbs G3B8 and Mab G3H2 which showed specific reactivity to purified r-PirB protein with no cross-reactivity to other shrimp bacterial pathogens (AHPND free Vibrio parahaemolyticus (Indian strains), V. anguillarum, WSSV, Aeromonas hydrophila, and Aphanomyces invadans). Immunodot developed using Mab G3B8 is more sensitive than that with the Mab G3H2. However, immunodot takes almost 2.5 hours to complete with several hands-on steps. Therefore, the flow-through assay (FTA) was developed by using a plastic cassette containing the nitrocellulose membrane with absorbing pads below. The sample was dotted in the test zone on the nitrocellulose membrane followed by continuos addition of five solutions in the order of i) blocking buffer (BSA) ii) primary antibody (MAb) iii) washing Solution iv) secondary antibody and v) chromogen substrate (TMB) clear purple dots against a white background were considered as positive reactions. The FTA developed using MAbG3B8 is more sensitive than that with MAb G3H2. In FTA the two MAbs showed specific reactivity to purified r-PirB protein and not to other shrimp bacterial pathogens. The FTA is simple to farmer/field level, sensitive and rapid requiring only 8-10 min for completion. Tests can be developed to kits, which will be ideal for use in biosecurity, for the first line of screening (at the port or pond site) and during monitoring and surveillance programmes overall for the good management practices to reduce the risk of the disease.

Keywords: acute hepatopancreatic necrosis disease, AHPND, flow-through assay, FTA, farmer level, immunodot, pond site, shrimp

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Authors: Brianne Nichols, John A. Parkinson

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Positive Psychology has provided a rich understanding of the beneficial effects of positive emotions in relation to optimal functioning, and research has been devoted to promote states of positive feeling and thinking. While this is a worthwhile pursuit, positive emotions are not useful in all contexts - some situations may require the individual to make use of their negative emotions to reach a desired end state. To account for the potential value of a wider range of emotional experiences that are common to the human condition, Positive Psychology needs to expand its horizons and investigate how individuals achieve positive outcomes using varied means. The current research seeks to understand the positive psychology of fear of failure (FF), which is a commonly experienced negative emotion relevant to most life domains. On the one hand, this emotion has been linked with avoidance motivation and self-handicap behaviours, on the other; FF has been shown to act as a drive to move the individual forward. To fully capture the depth of this highly subjective emotional experience and understand the circumstances under which FF may be adaptive, this study adopted a mixed methods design using SenseMaker; a web-based tool that combines the richness of narratives with the objectivity of numerical data. Two hundred participants consisting mostly of undergraduate university students shared a story of a time in the recent past when they feared failure of achieving a valued goal. To avoid researcher bias in the interpretation of narratives, participants self-signified their stories in a tagging system that was based on researchers’ aim to explore the role of past failures, the cognitive, emotional and behavioural profile of individuals high and low in FF, and the relationship between these factors. In addition, the role of perceived personal control and self-esteem were investigated in relation to FF using self-report questionnaires. Results from quantitative analyses indicated that individuals with high levels of FF, compared to low, were strongly influenced by past failures and preoccupied with their thoughts and emotions relating to the fear. This group also reported an unwillingness to accept their internal experiences, which in turn was associated with withdrawal from goal pursuit. Furthermore, self-esteem was found to mediate the relationship between perceived control and FF, suggesting that self-esteem, with or without control beliefs, may have the potential to buffer against high FF. It is hoped that the insights provided by the current study will inspire future research to explore the ways in which ‘acceptance’ may help individuals keep moving towards a goal despite the presence of FF, and whether cultivating a non-contingent self-esteem is the key to resilience in the face of failures.

Keywords: fear of failure, goal-pursuit, negative emotions, optimal functioning, resilience

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Authors: Michelle Belinda S. Weerawardana, Gobika Thiripuranathar, Priyani A. Paranagama

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Most of fresh vegetables and fruits available in the retail markets undergo a physiological disorder in its appearance and coloration, which indeed discourages consumer purchase. A loss of millions of dollars yearly to the food industry had been due to this pronounced color reaction called Enzymatic Browning which is driven due to the catalytic activity by an oxidoreductase enzyme, polyphenol oxidase (PPO). The enzyme oxidizes the phenolic compounds which are abundantly available in fruits and vegetables as substrates into quinones, which could react with proteins in its surrounding to generate black pigments, called melanins, which are highly UV-active compounds. Annona muricata (Katu anoda) and Musa acuminata (Ash plantains) is a fruit and a vegetable consumed by Sri Lankans widely due to their high nutritional values, medicinal properties and economical importance. The objective of the present study was to evaluate and determine the effective natural anti-browning inhibitors that could prevent PPO activity in the selected fruit and vegetable. Enzyme extracts from Annona muricata (Katu anoda) and Musa acuminata (Ash plantains), were prepared by homogenizing with analytical grade acetone, and pH of each enzyme extract was maintained at 7.0 using a phosphate buffer. The extracts of inhibitors were prepared using powdered ginger rhizomes and essential oil from the bark of Cinnamomum zeylanicum. Water extracts of ginger were prepared and the essential oil from Ceylon cinnamon bark was extracted using steam distillation method. Since the essential oil is not soluble in water, 0.1µl of cinnamon bark oil was mixed with 0.1µl of Triton X-100 emulsifier and 5.00 ml of water. The effect of each inhibitor on the PPO activity was investigated using catechol (0.1 mol dm-3) as the substrate and two samples of enzyme extracts prepared. The dosages of the prepared Cinnamon bark oil, and ginger (2 samples) which were used to measure the activity were 0.0035 g/ml, 0.091 g/ml and 0.087 g/ml respectively. The measurements of the inhibitory activity were obtained at a wavelength of 525 nm using the UV-visible spectrophotometer. The results evaluated thus revealed that % inhibition observed with cinnamon bark oil, and ginger for Annona muricata was 51.97%, and 60.90% respectively. The effects of cinnamon bark oil, and ginger extract on PPO activity of Musa acuminata were 49.51%, and 48.10%. The experimental findings thus revealed that Cinnamomum zeylanicum bark oil was a more effective inhibitor for PPO enzyme present in Musa acuminata and ginger was effective for PPO enzyme present in Annona muricata. Overall both the inhibitors were proven to be more effective towards the activities of PPO enzyme present in both samples. These inhibitors can thus be corroborated as effective, natural, non-toxic, anti-browning extracts, which when added to the above fruit and vegetable will increase the shelf life and also the acceptance of the product by the consumers.

Keywords: anti-browning agent, enzymatic browning, inhibitory activity, polyphenol oxidase

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Authors: Carlos A. C. G. Neto, Natan C. G. e Silva , Thaís de O. Costa, Luciana R. B. Gonçalves, Maria V. P. Rocha

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β-galactosidases (E.C. 3.2.1.23) are enzymes that have attracted by catalyzing the hydrolysis of lactose and in producing galacto-oligosaccharides by favoring transgalactosylation reactions. These enzymes, when immobilized, can have some enzymatic characteristics substantially improved, and the coating of supports with multifunctional polymers is a promising alternative to enhance the stability of the biocatalysts, among which polyethylenimine (PEI) stands out. PEI has certain properties, such as being a flexible polymer that suits the structure of the enzyme, giving greater stability, especially for multimeric enzymes such as β-galactosidases. Besides that, protects them from environmental variations. The use of chitosan support coated with PEI could improve the catalytic efficiency of β-galactosidase from Kluyveromyces lactis in the transgalactosylation reaction for the production of prebiotics, such as lactulose since this strain is more effective in the hydrolysis reaction. In this context, the aim of the present work was first to develop biocatalysts of β-galactosidase from K. lactis immobilized on chitosan-coated with PEI, determining the immobilization parameters, its operational and thermal stability, and then to apply it in hydrolysis and transgalactolisation reactions to produce lactulose using whey as a substrate. The immobilization of β-galactosidase in chitosan previously functionalized with 0.8% (v/v) glutaraldehyde and then coated with 10% (w/v) PEI solution was evaluated using an enzymatic load of 10 mg protein per gram support. Subsequently, the hydrolysis and transgalactosylation reactions were conducted at 50 °C, 120 RPM for 20 minutes, using whey supplemented with fructose at a ratio of 1:2 lactose/fructose, totaling 200 g/L. Operational stability studies were performed in the same conditions for 10 cycles. Thermal stabilities of biocatalysts were conducted at 50 ºC in 50 mM phosphate buffer, pH 6.6 with 0.1 mM MnCl2. The biocatalyst whose support was coated was named CHI_GLU_PEI_GAL, and the one that was not coated was named CHI_GLU_GAL. The coating of the support with PEI considerably improved the parameters of immobilization. The immobilization yield increased from 56.53% to 97.45%, biocatalyst activity from 38.93 U/g to 95.26 U/g and the efficiency from 3.51% to 6.0% for uncoated and coated support, respectively. The biocatalyst CHI_GLU_PEI_GAL was better than CHI_GLU_GAL in the hydrolysis of lactose and production of lactulose, converting 97.05% of lactose at 5 min of reaction and producing 7.60 g/L lactulose in the same time interval. QUI_GLU_PEI_GAL biocatalyst was stable in the hydrolysis reactions of lactose during the 10 cycles evaluated, converting 73.45% lactose even after the tenth cycle, and in the lactulose production was stable until the fifth cycle evaluated, producing 10.95 g/L lactulose. However, the thermal stability of CHI_GLU_GAL biocatalyst was superior, with a half-life time 6 times higher, probably because the enzyme was immobilized by covalent bonding, which is stronger than adsorption (CHI_GLU_PEI_GAL). Therefore, the strategy of coating the supports with PEI has proven to be effective for the immobilization of β-galactosidase from K. lactis, considerably improving the immobilization parameters, as well as, the catalytic action of the enzyme. Besides that, this process can be economically viable due to the use of an industrial residue as a substrate.

Keywords: β-galactosidase, immobilization, kluyveromyces lactis, lactulose, polyethylenimine, transgalactosylation reaction, whey

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Authors: Carlos A. C. G. Neto, Natan C. G. Silva, Thais O. Costa, Luciana R. B. Goncalves, Maria v. P. Rocha

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Galactosidases are enzymes responsible for catalyzing lactose hydrolysis reactions and also favoring transgalactosylation reactions for the production of prebiotics, among which lactulose stands out. These enzymes, when immobilized, can have some enzymatic characteristics substantially improved, and the coating of supports with multifunctional polymers in immobilization processes is a promising alternative in order to extend the useful life of the biocatalysts, for example, the coating with polyethyleneimine (PEI). PEI is a flexible polymer that suits the structure of the enzyme, giving greater stability, especially for multimeric enzymes such as β-galactosidases and also protects it from environmental variations, for example, pH and temperature. In addition, it can substantially improve the immobilization parameters and also the efficiency of enzymatic reactions. In this context, the aim of the present work was first to develop biocatalysts of β-galactosidase from Kluyveromyces lactis immobilized on PEI coated agarose, determining the immobilization parameters, its operational and thermal stability, and then to apply it in the hydrolysis of lactose and synthesis of lactulose, using whey as a substrate. This immobilization strategy was chosen in order to improve the catalytic efficiency of the enzyme in the transgalactosylation reaction for the production of prebiotics, and there are few studies with β-galactosidase from this strain. The immobilization of β-galactosidase in agarose previously functionalized with 48% (w/v) glycidol and then coated with 10% (w/v) PEI solution was evaluated using an enzymatic load of 10 mg/g of protein. Subsequently, the hydrolysis and transgalactosylation reactions were conducted at 50 °C, 120 RPM for 20 minutes, using whey (66.7 g/L of lactose) supplemented with 133.3 g/L fructose at a ratio of 1:2 (lactose/fructose). Operational stability studies were performed in the same conditions for 10 cycles. Thermal stabilities of biocatalysts were conducted at 50 ºC in 50 mM phosphate buffer, pH 6.6, with 0.1 mM MnCl2. The biocatalysts whose supports were coated were named AGA_GLY_PEI_GAL, and those that were not coated were named AGA_GLY_GAL. The coating of the support with PEI considerably improved immobilization yield (2.6-fold), the biocatalyst activity (1.4-fold), and efficiency (2.2-fold). The biocatalyst AGA_GLY_PEI_GAL was better than AGA_GLY_GAL in hydrolysis and transgalactosylation reactions, converting 88.92% of lactose at 5 min of reaction and obtaining a residual concentration of 5.24 g/L. Besides that, it was produced 13.90 g/L lactulose in the same time interval. AGA_GLY_PEI_GAL biocatalyst was stable during the 10 cycles evaluated, converting approximately 80% of lactose and producing 10.95 g/L of lactulose even after the tenth cycle. However, the thermal stability of AGA_GLY_GAL biocatalyst was superior, with a half-life time 5 times higher, probably because the enzyme was immobilized by covalent bonding, which is stronger than adsorption (AGA_GLY_PEI_GAL). Therefore, the strategy of coating the supports with PEI has proven to be effective for the immobilization of β-galactosidase from K. lactis, considerably improving the immobilization parameters, as well as the enzyme, catalyzed reactions. In addition, the use of whey as a raw material for lactulose production has proved to be an industrially advantageous alternative.

Keywords: β-galactosidase, immobilization, lactulose, polyethylenimine, whey

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Authors: Katarzyna Bialik-Wąs, Klaudia Pluta, Dagmara Malina, Małgorzata Miastkowska

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In recent years, biocompatible hydrogels have been used more and more in medical applications, especially as modern dressings and drug delivery systems. The main goal of this research was the characteristics of bio-hybrid hydrogel materials incorporated with the nanocarrier-drug system, which enable the release in a gradual and prolonged manner, up to 7 days. Therefore, the use of such a combination will provide protection against mechanical damage and adequate hydration. The proposed bio-hybrid hydrogels are characterized by: transparency, biocompatibility, good mechanical strength, and the dual release system, which allows for gradual delivery of the active substance, even up to 7 days. Bio-hybrid hydrogels based on sodium alginate (SA), poly(vinyl alcohol) (PVA), glycerine, and Aloe vera solution (AV) were obtained through the chemical crosslinking method using poly(ethylene glycol) diacrylate as a crosslinking agent. Additionally, a nanocarrier-drug system was incorporated into SA/PVA/AV hydrogel matrix. Here, studies were focused on the release profiles of active substances from bio-hybrid hydrogels using the USP4 method (DZF II Flow-Through System, Erweka GmbH, Langen, Germany). The equipment incorporated seven in-line flow-through diffusion cells. The membrane was placed over support with an orifice of 1,5 cm in diameter (diffusional area, 1.766 cm²). All the cells were placed in a cell warmer connected with the Erweka heater DH 2000i and the Erweka piston pump HKP 720. The piston pump transports the receptor fluid via seven channels to the flow-through cells and automatically adapts the setting of the flow rate. All volumes were measured by gravimetric methods by filling the chambers with Milli-Q water and assuming a density of 1 g/ml. All the determinations were made in triplicate for each cell. The release study of the model active substance was carried out using a regenerated cellulose membrane Spectra/Por®Dialysis Membrane MWCO 6-8,000 Carl Roth® Company. These tests were conducted in buffer solutions – PBS at pH 7.4. A flow rate of receptor fluid of about 4 ml /1 min was selected. The experiments were carried out for 7 days at a temperature of 37°C. The released concentration of the model drug in the receptor solution was analyzed using UV-Vis spectroscopy (Perkin Elmer Company). Additionally, the following properties of the modified materials were studied: physicochemical, structural (FT-IR analysis), morphological (SEM analysis). Finally, the cytotoxicity tests using in vitro method were conducted. The obtained results exhibited that the dual release system allows for the gradual and prolonged delivery of the active substances, even up to 7 days.

Keywords: wound dressings, SA/PVA hydrogels, nanocarrier-drug system, USP4 method

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Authors: Marco Ranzato, Maguelone Vignes

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The co-design process recently run by the trans-disciplinary urban laboratory Metrolab Brussels for outlining the architecture of a future integrated health centre in Brussels (Belgium) has highlighted that a buffer place open to the local community is the appropriate cornerstone around which organizing a space where diverse professionals and patients are together. In the context of the migrants 'crisis' in Europe, the growing number of vulnerable people in Brussels and the increasing complexity of the health and welfare systems, the NGO Doctors of the World (DoW) has launched a project funded by The European Regional Development Fund, and aiming to create a new community centre combining social and health services in a poor but changing neighborhood of Brussels. Willing not to make a 'ghetto' of this new integrated service, the NGO looks at hosting different publics in order to make the poorest, marginal and most vulnerable people access to a regular kind of service. As a trans-disciplinary urban research group, Metrolab has been involved in the process of co-designing the architecture of the future centre with a set of various health professionals, social workers, and patients’ representatives. Metrolab drawn on the participants’ practice experiences and knowledge of hosting different kinds of publics and professions in a same structure in order to imagine what rooms should fit into the centre, what atmosphere they should convey, how should they be interrelated and organized, and, concurrently, how the building should fit into the urban frame of its neighborhood. The result is that, in order for an integrated health centre framed in the landscape of a disadvantaged neighborhood to function, it has to work as social community centre offering accessibility and conviviality to diverse social groups. This paper outlines the methodology that Metrolab used to design and conduct, in close collaboration with DoW, a series of 3 workshops. Through sketching and paper modeling, the methodology made participants talk about their experience by projecting them into a situation. It included a combination of individual and collective work in order to sharp participants’ eyes on architectural forms, explicit their thoughts and experience through inter-subjectivity and imagine solutions to the challenges they raised. Such a collaborative method encompasses several challenges about patients’ participation and representation, replicability of the conditions of success and the plurality of the research findings communication formats. This paper underlines how this participatory process has contributed to build knowledge on the few-documented topic of the architecture of community health centres. More importantly, the contribution builds on this participatory process to discuss the importance of adapting the architecture of the new integrated health centre to the changing population of Brussels and to the issues of its specific neighborhood.

Keywords: co-design, health, social innovation, urban lab

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Authors: Busra Gundede, Ozal Mutlu, Nagihan Gulsoy

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Background: Over the years, material research has led to the development of numerous metals and alloys for using in biomedical applications. One of the major tasks of biomaterial research is the functionalization of the material surface to improve the biocompatibility according to a specific application. 316L and 316L alloys are excellent for various bio-applications. This research was investigated the effect of titanium (Ti), niobium (Nb), and zirconium (Zr) additives on injection molded austenitic grade 316L stainless steels in vitro biocompatibility. For this purpose, cytotoxic tests were performed to evaluate the potential biocompatibility of the specimens. Materials and Methods: 3T3 fibroblast were cultivated in DMEM supplemented with 10% fetal bovine serum and %1 penicillin-streptomycin at 37°C with 5% CO2 and 95%humidity. Trypsin/EDTA solution was used to remove cells from the culture flask. Cells were reseeded at a density of 1×105cell in 25T flasks. The medium change took place every 3 days. The trypan blue assay was used to determine cell viability. Cell viability is calculated as the number of viable cells divided by the total number of cells within the grids on the cell counter machine counted the number of blue staining cells and the number of total cells. Cell viability should be at least 95% for healthy log-phase cultures. MTT assay was assessed for 96-hours. Cells were cultivated in 6-well flask within 5 ml DMEM and incubated as same conditions. 0,5mg/ml MTT was added for 4-hours and then acid-isoprohanol was added for solubilize to formazan crystals. Cell morphology after 96h was investigated by SEM. The medium was removed, samples were washed with 0.15 M PBS buffer and fixed for 12h at 4- 8°C with %2,5 gluteraldehyte. Samples were treated with 1% osmium tetroxide. Samples were then dehydrated and dried, mounted on appropriate stubs with colloidal silver and sputter-coated with gold. Images were collected using a scanning electron microscope. ROS assay is a cell viability test for in vitro studies. Cells were grown for 96h, ROS solution added on cells in 6 well plate flask and incubated for 1h. Fluorescence signal indicates ROS generation by cells. Results: Trypan Blue exclusion assay results were 96%, 92%, 95%, 90%, 91% for negative control group, 316L, 316L-Ti, 316L-Nb and 316L-Zr, respectively. Results were found nearly similar to each other when compared with control group. Cell viability from MTT analysis was found to be 100%, 108%, 103%, 107%, and 105% for the control group, 316L, 316L-Ti, 316L-Nb and 316L-Zr, respectively. Fluorescence microscopy analysis indicated that all test groups were same as the control group in ROS assay. SEM images demonstrated that the attachment of 3T3 cells on biomaterials. Conclusion: We, therefore, concluded that Ti, Nb and Zr additives improved physical properties of 316L stainless. In our in vitro experiments showed that these new additives did not modify the cytocompatibility of stainless steel and these additives on 316L might be useful for biomedical applications.

Keywords: 316L stainles steel, biocompatibility, cell culture, Ti, Nb, Zr

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Authors: Yana Posmitnaya, Galina Rudnitskaya, Tatyana Lukashenko, Anton Bukatin, Anatoly Evstrapov

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An important area of biological and medical research is the study of genetic mutations and polymorphisms that can alter gene function and cause inherited diseases and other diseases. The following methods to analyse DNA fragments are used: capillary electrophoresis and electrophoresis on microfluidic chip (MFC), mass spectrometry with electrophoresis on MFC, hybridization assay on microarray. Electrophoresis on MFC allows to analyse small volumes of samples with high speed and throughput. A soft lithography in polydimethylsiloxane (PDMS) was chosen for operative fabrication of MFCs. A master-form from silicon and photoresist SU-8 2025 (MicroChem Corp.) was created for the formation of micro-sized structures in PDMS. A universal topology which combines T-injector and simple cross was selected for the electrophoretic separation of the sample. Glass K8 and PDMS Sylgard® 184 (Dow Corning Corp.) were used for fabrication of MFCs. Electroosmotic flow (EOF) plays an important role in the electrophoretic separation of the sample. Therefore, the estimate of the quantity of EOF and the ways of its regulation are of interest for the development of the new methods of the electrophoretic separation of biomolecules. The following methods of surface modification were chosen to change EOF: high-frequency (13.56 MHz) plasma treatment in oxygen and argon at low pressure (1 mbar); 1% aqueous solution of polyvinyl alcohol; 3% aqueous solution of Kolliphor® P 188 (Sigma-Aldrich Corp.). The electroosmotic mobility was evaluated by the method of Huang X. et al., wherein the borate buffer was used. The influence of physical and chemical methods of treatment on the wetting properties of the PDMS surface was controlled by the sessile drop method. The most effective way of surface modification of MFCs, from the standpoint of obtaining the smallest value of the contact angle and the smallest value of the EOF, was the processing with aqueous solution of Kolliphor® P 188. This method of modification has been selected for the treatment of channels of MFCs, which are used for the separation of mixture of oligonucleotides fluorescently labeled with the length of chain with 10, 20, 30, 40 and 50 nucleotides. Electrophoresis was performed on the device MFAS-01 (IAI RAS, Russia) at the separation voltage of 1500 V. 6% solution of polydimethylacrylamide with the addition of 7M carbamide was used as the separation medium. The separation time of components of the mixture was determined from electropherograms. The time for untreated MFC was ~275 s, and for the ones treated with solution of Kolliphor® P 188 – ~ 220 s. Research of physical-chemical methods of surface modification of MFCs allowed to choose the most effective way for reducing EOF – the modification with aqueous solution of Kolliphor® P 188. In this case, the separation time of the mixture of oligonucleotides decreased about 20%. The further optimization of method of modification of channels of MFCs will allow decreasing the separation time of sample and increasing the throughput of analysis.

Keywords: electrophoresis, microfluidic chip, modification, nucleic acid, polydimethylsiloxane, soft lithography

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Authors: Norihiro Kato, Yuriko Takayama

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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

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Authors: Zafar Tabrez, Nizamuddin Khan

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Agriculture in India in the pre-green revolution period was mostly controlled by terrain, climate and edaphic factors. But after the introduction of innovative factors and technological inputs, green revolution occurred and agricultural scene witnessed great change. In the development of India’s agriculture, speedy, and extensive introduction of technological change is one of the crucial factors. The technological change consists of adoption of farming techniques such as use of fertilisers, pesticides and fungicides, improved variety of seeds, modern agricultural implements, improved irrigation facilities, contour bunding for the conservation of moisture and soil, which are developed through research and calculated to bring about diversification and increase of production and greater economic return to the farmers. The green revolution in India took place during late 60s, equipped with technological inputs like high yielding varieties seeds, assured irrigation as well as modern machines and implements. Initially the revolution started in Punjab, Haryana and western Uttar Pradesh. With the efforts of government, agricultural planners, as well as policy makers, the modern technocratic agricultural development scheme was also implemented and introduced in backward and marginal regions of the country later on. Agriculture sector occupies the centre stage of India’s social security and overall economic welfare. The country has attained self-sufficiency in food grain production and also has sufficient buffer stock. Our first Prime Minister, Jawaharlal Nehru said ‘everything else can wait but not agriculture’. There is still a continuous change in the technological inputs and cropping patterns. Keeping these points in view, author attempts to investigate extensively the mechanization of agriculture and the change by selecting western Trans-Ghaghara plain as a case study and block a unit of the study. It includes the districts of Gonda, Balrampur, Bahraich and Shravasti which incorporate 44 blocks. It is based on secondary sources of data by blocks for the year 1997 and 2007. It may be observed that there is a wide range of variations and the change in farm mechanization, i.e., agricultural machineries such as ploughs, wooden and iron, advanced harrow and cultivator, advanced thrasher machine, sprayers, advanced sowing instrument, and tractors etc. It may be further noted that due to continuous decline in size of land holdings and outflux of people for the same nature of works or to be employed in non-agricultural sectors, the magnitude and direction of agricultural systems are affected in the study area which is one of the marginalized regions of Uttar Pradesh, India.

Keywords: agriculture, technological inputs, farm mechanization, food production, cropping pattern

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Authors: Yuriko Takayama, Norihiro Kato

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Quorum sensing (QS) is a cell-to-cell communication system in bacteria to regulate expression of target genes. In gram-negative bacteria, activation on QS is controlled by a concentration increase of N-acylhomoserine lactone (AHL), which can diffuse in and out of the cell. Effective control of QS is expected to avoid virulence factor production in infectious pathogens, biofilm formation, and antibiotic production because various cell functions in gram-negative bacteria are controlled by AHL-mediated QS. In this research, we applied cyclodextrins (CDs) as artificial hosts for the AHL signal to reduce the AHL concentration in the culture broth below its threshold for QS activation. The AHL-receptor complex induced under the high AHL concentration activates transcription of the QS-target gene. Accordingly, artificial reduction of the AHL concentration is one of the effective strategies to inhibit the QS. A hydrophobic cavity of the CD can interact with the acyl-chain of the AHL due to hydrophobic interaction in aqueous media. We studied N-hexanoylhomoserine lactone (C6HSL)-mediated QS in Serratia marcescens; accumulation of C6HSL is responsible for regulation of the expression of pig cluster. Inhibitory effects of added CDs on QS were demonstrated by determination of prodigiosin amount inside cells after reaching stationary phase, because production of prodigiosin depends on the C6HSL-mediated QS. By adding approximately 6 wt% hydroxypropyl-β-CD (HP-β-CD) in Luria-Bertani (LB) medium prior to inoculation of S. maecescens AS-1, the intracellularly accumulated prodigiosin was drastically reduced to 7-10%, which was determined after the extraction of prodigiosin in acidified ethanol. The AHL retention ability of HP-β-CD was also demonstrated by Chromobacterium violacuem CV026 bioassay. The CV026 strain is an AHL-synthase defective mutant that activates QS solely by adding AHLs from outside of cells. A purple pigment violacein is induced by activation of the AHL-mediated QS. We demonstrated that the violacein production was effectively suppressed when the C6HSL standard solution was spotted on a LB agar plate dispersing CV026 cells and HP-β-CD. Physico-chemical analysis was performed to study the affinity between the immobilized CD and added C6HSL using a quartz crystal microbalance (QCM) sensor. The COOH-terminated self-assembled monolayer was prepared on a gold electrode of 27-MHz AT-cut quartz crystal. Mono(6-deoxy-6-N, N-diethylamino)-β-CD was immobilized on the electrode using water-soluble carbodiimide. The C6HSL interaction with the β-CD cavity was studied by injecting the C6HSL solution to a cup-type sensor cell filled with buffer solution. A decrement of resonant frequency (ΔFs) clearly showed the effective C6HSL complexation with immobilized β-CD and its stability constant for MBP-SpnR-C6HSL complex was on the order of 102 M-1. The CD has high potential for engineered control of QS because it is safe for human use.

Keywords: acylhomoserine lactone, cyclodextrin, intracellular signaling, quorum sensing

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Authors: Thauane Silva, Gabrielle do Vale, Andre Ferreira, Marilda Siqueira, Thiago Moreno L. Souza, Milene D. Miranda

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The exposure of A(H1N1)pdm09-infected epithelial cells (HeLa) to HIV-1 viral particles, or its gp120, enhanced interferon-induced transmembrane protein (IFITM3) content, a viral restriction factor (RF), resulting in a decrease in influenza replication. The gp120 binds to CCR5 (R5) or CXCR4 (X4) cell receptors during HIV-1 infection. Then, it is possible that the endogenous ligands of these receptors also modulate the expression of IFITM3 and other cellular factors that restrict influenza virus replication. Thus, the aim of this study is to analyze the role of cellular receptors R5 and X4 in modulating RFs in order to inhibit the replication of the influenza virus. A549 cells were treated with 2x effective dose (ED50) of endogenous R5 or X4 receptor agonists, CCL3 (20 ng/ml), CCL4 (10 ng/ml), CCL5 (10 ng/ml) and CXCL12 (100 ng/mL) or exogenous agonists, gp120 Bal-R5, gp120 IIIB-X4 and its mutants (5 µg/mL). The interferon α (10 ng/mL) and oseltamivir (60 nM) were used as a control. After 24 h post agonists exposure, the cells were infected with virus influenza A(H3N2) at 2 MOI (multiplicity of infection) for 1 h. Then, 24 h post infection, the supernatant was harvested and, the viral titre was evaluated by qRT-PCR. To evaluate IFITM3 and SAM and HD domain containing deoxynucleoside triphosphate triphosphohydrolase 1 (SAMHD1) protein levels, A549 were exposed to agonists for 24 h, and the monolayer was lysed with Laemmli buffer for western blot (WB) assay or fixed for indirect immunofluorescence (IFI) assay. In addition to this, we analyzed other RFs modulation in A549, after 24 h post agonists exposure by customized RT² Profiler Polymerase Chain Reaction Array. We also performed a functional assay in which SAMHD1-knocked-down, by single-stranded RNA (siRNA), A549 cells were infected with A(H3N2). In addition, the cells were treated with guanosine to assess the regulatory role of dNTPs by SAMHD1. We found that R5 and X4 agonists inhibited influenza replication in 54 ± 9%. We observed a four-fold increase in SAMHD1 transcripts by RFs mRNA quantification panel. After 24 h post agonists exposure, we did not observe an increase in IFITM3 protein levels through WB or IFI assays, but we observed an upregulation up to three-fold in the protein content of SAMHD1, in A549 exposed to agonists. Besides this, influenza replication enhanced in 20% in cell cultures that SAMDH1 was knockdown. Guanosine treatment in cells exposed to R5 ligands further inhibited influenza virus replication, suggesting that the inhibitory mechanism may involve the activation of the SAMHD1 deoxynucleotide triphosphohydrolase activity. Thus, our data show for the first time a direct relationship of SAMHD1 and inhibition of influenza replication, and provides perspectives for new studies on the signaling modulation, through cellular receptors, to induce proteins of great importance in the control of relevant infections for public health.

Keywords: chemokine receptors, gp120, influenza, virus restriction factors

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Authors: Shanlong Lu, Jiming Jin, Xiaochun Wang

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The Tibetan Plateau has the largest number of inland lakes with the highest elevation on the planet. These massive and large lakes are mostly in natural state and are less affected by human activities. Their shrinking or expansion can truly reflect regional climate and environmental changes and are sensitive indicators of global climate change. However, due to the sparsely populated nature of the plateau and the poor natural conditions, it is difficult to effectively obtain the change data of the lake, which has affected people's understanding of the temporal and spatial processes of lake water changes and their influencing factors. By using the MODIS (Moderate Resolution Imaging Spectroradiometer) MOD09Q1 surface reflectance images as basic data, this study produced the 8-day lake water surface data set of the Tibetan Plateau from 2000 to 2012 at 250 m spatial resolution, with a lake water surface extraction method of combined with lake water surface boundary buffer analyzing and lake by lake segmentation threshold determining. Then based on the dataset, the lake water surface variations and their influencing factors were analyzed, by using 4 typical natural geographical zones of Eastern Qinghai and Qilian, Southern Qinghai, Qiangtang, and Southern Tibet, and the watersheds of the top 10 lakes of Qinghai, Siling Co, Namco, Zhari NamCo, Tangra Yumco, Ngoring, UlanUla, Yamdrok Tso, Har and Gyaring as the analysis units. The accuracy analysis indicate that compared with water surface data of the 134 sample lakes extracted from the 30 m Landsat TM (Thematic Mapper ) images, the average overall accuracy of the lake water surface data set is 91.81% with average commission and omission error of 3.26% and 5.38%; the results also show strong linear (R2=0.9991) correlation with the global MODIS water mask dataset with overall accuracy of 86.30%; and the lake area difference between the Second National Lake Survey and this study is only 4.74%, respectively. This study provides reliable dataset for the lake change research of the plateau in the recent decade. The change trends and influencing factors analysis indicate that the total water surface area of lakes in the plateau showed overall increases, but only lakes with areas larger than 10 km2 had statistically significant increases. Furthermore, lakes with area larger than 100 km2 experienced an abrupt change in 2005. In addition, the annual average precipitation of Southern Tibet and Southern Qinghai experienced significant increasing and decreasing trends, and corresponding abrupt changes in 2004 and 2006, respectively. The annual average temperature of Southern Tibet and Qiangtang showed a significant increasing trend with an abrupt change in 2004. The major reason for the lake water surface variation in Eastern Qinghai and Qilian, Southern Qinghai and Southern Tibet is the changes of precipitation, and that for Qiangtang is the temperature variations.

Keywords: lake water surface variation, MODIS MOD09Q1, remote sensing, Tibetan Plateau

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Authors: Arinze Ezieke, Antonio Serrano, William Clarke, Denys Villa-Gomez

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Berry cultivation results in discharge of high organic strength putrescible solid waste which potentially contributes to environmental degradation, making it imperative to assess options for its complete management. Anaerobic digestion (AD) could be an ideal option when the target is energy generation; however, due to berry fruit characteristics high carbohydrate composition, the technology could be limited by its high alkalinity requirement which suggests dosing of additives such as buffers and trace elements supplement. Overcoming this limitation in an economically viable way could entail replacement of synthetic additives with recycled by-product waste. Consequently, ash from co-smouldering of high COD characteristic AD digestate and coco-coir could be a promising material to be used to enhance the AD of berry fruit waste, given its characteristic high pH, alkalinity and metal concentrations which is typical of synthetic additives. Therefore, the aim of the research was to evaluate the stability and process performance from the AD of BFW when ash from co-smoldered digestate and coir are supplemented as alkalinity and trace elements (TEs) source. Series of batch experiments were performed to ascertain the necessity for alkalinity addition and to see whether the alkalinity and metals in the co-smouldered digestate ash can provide the necessary buffer and TEs for AD of berry fruit waste. Triplicate assays were performed in batch systems following I/S of 2 (in VS), using serum bottles (160 mL) sealed and placed in a heated room (35±0.5 °C), after creating anaerobic conditions. Control experiment contained inoculum and substrates only, and inoculum, substrate and NaHCO3 for optimal total alkalinity concentration and TEs assays, respectively. Total alkalinity concentration refers to alkalinity of inoculum and the additives. The alkalinity and TE potential of the ash were evaluated by supplementing ash (22.574 g/kg) of equivalent total alkalinity concentration to that of the pre-determined optimal from NaHCO3, and by dosing ash (0.012 – 7.574 g/kg) of varying concentrations of specific essential TEs (Co, Fe, Ni, Se), respectively. The result showed a stable process at all examined conditions. Supplementation of 745 mg/L CaCO3 NaHCO3 resulted to an optimum TAC of 2000 mg/L CaCO3. Equivalent ash supplementation of 22.574 g/kg allowed the achievement of this pre-determined optimum total alkalinity concentration, resulting to a stable process with a 92% increase in the methane production rate (323 versus 168 mL CH4/ (gVS.d)), but a 36% reduction in the cumulative methane production (103 versus 161 mL CH4/gVS). Addition of ashes at incremental dosage as TEs source resulted to a reduction in the Cumulative methane production, with the highest dosage of 7.574 g/kg having the highest effect of -23.5%; however, the seemingly immediate bioavailability of TE at this high dosage allowed for a +15% increase in the methane production rate. With an increased methane production rate, the results demonstrated that the ash at high dosages could be an effective supplementary material for either a buffered or none buffered berry fruit waste AD system.

Keywords: anaerobic digestion, alkalinity, co-smoldered digestate ash, trace elements

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Authors: Jabulani Gilford Kheswa, Thembelihle Lobi

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From a cultural perspective, expression of hegemonic masculinity in South Africa continues to escalate among adolescent males who grow up in communities lacking in role models and recreational facilities. However, when the schools are constructive, and peer influence is positive, adolescent male can potentially express character strengths and lead a meaningful life. Drawing from Bronfenbrenner’s Ecological Model and Keyes and Ryff’s six dimensions of psychological well-being and mental health, such youth may exemplify positive self-esteem, problem- focused coping strategies, condom self-efficacy, good leadership skills, enhanced motivation and a positive emotional state, which buffer against risky sexual behaviors. This paper was aimed at investigating the relationships between adolescent males’ sexual behavior and psychological well-being. This study employed a quantitative research to collect data from 54 Xhosa-speaking adolescent males from one school high school in Fort Beaufort, Eastern Cape, South Africa. These learners were from grade nine, ten and eleven with their ages ranging from 14 to 20. Prior the research commenced, the school principal and caregivers of the learners who participated in the study, gave their informed consent. Self- administered closed-ended questionnaire with Section A (that is, biographical information) and Section B with each question rated on the 5–point Likert scale was used. The advantages of questionnaires include a high response rate as they require less time and offer anonymity because participants’ names are not identified. The SPSS version 18 was used for statistical data analysis. The mean age was 16.83 with a standard deviation of 1.611. 44.4% of the participants were from grade 9, 33.3% from grade 10 and 22.2% from grade 11. The Chronbach alpha of 0.79 was yielded, with respect to self- esteem of adolescent males. In this study, 76.9% reported to attend church services whilst 23% indicated not to attend church services. A further 96.2% of adolescent males indicated to have good relations with guardians while only 3.8% had poorer relations. A large proportion of adolescent males (72.9%) indicated to high-quality friendship as opposed to 27.1% who reported being receiving negative guidance from peers. Other findings revealed that 81.1% of the participants’ parents do not drink alcohol, and they cope at school as 79.6% reported protective factors as attributable towards non-engagement to risky sexual practices. As a result, 81.4% of participants reported not to participate in criminal activities although 85% of the participants indicated that in their school there are drugs. It could be speculated from this study that adolescent males whose caregivers are authoritative, find purpose in life and are most likely to be socially and academically competent. This paper leads to further research interest into mental health, coping strategies and sexual decision-making skills of the youth in South Africa.

Keywords: church, mental health, school, sexual behaviour

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Authors: Corentin Jankowiak, Aggelos Zacharopoulos, Caterina Brandoni

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In a context of increasing stress put on the electricity network by the decarbonization of many sectors, energy storage is likely to be the key mitigating element, by acting as a buffer between production and demand. In particular, the highest potential for storage is when connected closer to the loads. Yet, low voltage storage struggles to penetrate the market at a large scale due to the novelty and complexity of the solution, and the competitive advantage of fossil fuel-based technologies regarding regulations. Strong and reliable numerical simulations are required to show the benefits of storage located near loads and promote its development. The present study was restrained from excluding aggregated control of storage: it is assumed that the storage units operate independently to one another without exchanging information – as is currently mostly the case. A computationally light battery model is presented in detail and validated by direct comparison with a domestic battery operating in real conditions. This model is then used to develop Peak-Shaving (PS) control strategies as it is the decentralized service from which beneficial impacts are most likely to emerge. The aggregation of flatter, peak- shaved consumption profiles is likely to lead to flatter and arbitraged profile at higher voltage layers. Furthermore, voltage fluctuations can be expected to decrease if spikes of individual consumption are reduced. The crucial part to achieve PS lies in the charging pattern: peaks depend on the switching on and off of appliances in the dwelling by the occupants and are therefore impossible to predict accurately. A performant PS strategy must, therefore, include a smart charge recovery algorithm that can ensure enough energy is present in the battery in case it is needed without generating new peaks by charging the unit. Three categories of PS algorithms are introduced in detail. First, using a constant threshold or power rate for charge recovery, followed by algorithms using the State Of Charge (SOC) as a decision variable. Finally, using a load forecast – of which the impact of the accuracy is discussed – to generate PS. A performance metrics was defined in order to quantitatively evaluate their operating regarding peak reduction, total energy consumption, and self-consumption of domestic photovoltaic generation. The algorithms were tested on load profiles with a 1-minute granularity over a 1-year period, and their performance was assessed regarding these metrics. The results show that constant charging threshold or power are far from optimal: a certain value is not likely to fit the variability of a residential profile. As could be expected, forecast-based algorithms show the highest performance. However, these depend on the accuracy of the forecast. On the other hand, SOC based algorithms also present satisfying performance, making them a strong alternative when the reliable forecast is not available.

Keywords: decentralised control, domestic integrated batteries, electricity network performance, peak-shaving algorithm

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Authors: Mayoorika Shukla, Pramila Jakhar, Tejendra Dixit, I. A. Palani, Vipul Singh

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Biosensors are analytical devices with wide range of applications in biological, chemical, environmental and clinical analysis. It comprises of bio-recognition layer which has biomolecules (enzymes, antibodies, DNA, etc.) immobilized over it for detection of analyte and transducer which converts the biological signal into the electrical signal. The performance of biosensor primarily the depends on the bio-recognition layer and therefore it has to be chosen wisely. In this regard, nanostructures of metal oxides such as ZnO, SnO2, V2O5, and TiO2, etc. have been explored extensively as bio-recognition layer. Recently, ZnO has the attracted attention of researchers due to its unique properties like high iso-electric point, biocompatibility, stability, high electron mobility and high electron binding energy, etc. Although there have been many reports on usage of ZnO as bio-recognition layer but to the authors’ knowledge, none has ever observed correlation between optical properties like defect suppression and biosensing capability of the sensor. Here, ZnO nanorods (ZNR) have been synthesized by a low cost, simple and low-temperature hydrothermal growth process, over Platinum (Pt) coated glass substrate. The ZNR have been synthesized in two steps viz. initially a seed layer was coated over substrate (Pt coated glass) followed by immersion of it into nutrient solution of Zinc nitrate and Hexamethylenetetramine (HMTA) with in situ addition of KMnO4. The addition of KMnO4 was observed to have a profound effect over the growth rate anisotropy of ZnO nanostructures. Clustered and powdery growth of ZnO was observed without addition of KMnO4, although by addition of it during the growth, uniform and crystalline ZNR were found to be grown over the substrate. Moreover, the same has resulted in suppression of defects as observed by Normalized Photoluminescence (PL) spectra since KMnO4 is a strong oxidizing agent which provides an oxygen rich growth environment. Further, to explore the correlation between defect suppression and biosensing capability of the ZNR Glucose oxidase (Gox) was immobilized over it, using physical adsorption technique followed by drop casting of nafion. Here the main objective of the work was to analyze effect of defect suppression over biosensing capability, and therefore Gox has been chosen as model enzyme, and electrochemical amperometric glucose detection was performed. The incorporation of KMnO4 during growth has resulted in variation of optical and charge transfer properties of ZNR which in turn were observed to have deep impact on biosensor figure of merits. The sensitivity of biosensor was found to increase by 12-18 times, due to variations introduced by addition of KMnO4 during growth. The amperometric detection of glucose in continuously stirred buffer solution was performed. Interestingly, defect suppression has been observed to contribute towards the improvement of biosensor performance. The detailed mechanism of growth of ZNR along with the overall influence of defect suppression on the sensing capabilities of the resulting enzymatic electrochemical biosensor and different figure of merits of the biosensor (Glass/Pt/ZNR/Gox/Nafion) will be discussed during the conference.

Keywords: biosensors, defects, KMnO4, ZnO nanorods

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Authors: D. P. Houhoula, J. Papaparaskevas, S. Konteles, A. Dargenta, A. Farka, C. Spyrou, M. Ziaka, S. Koussisis, E. Charvalos

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Objectives: Nanotechnology is providing revolutionary opportunities for the rapid and simple diagnosis of many infectious diseases. Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis are important human pathogens. Diagnostic assays for bacterial culture and identification are time consuming and laborious. There is an urgent need to develop rapid, sensitive, and inexpensive diagnostic tests. In this study, a gold nanoprobe strategy developed and relies on the colorimetric differentiation of specific DNA sequences based approach on differential aggregation profiles in the presence or absence of specific target hybridization. Method: Gold nanoparticles (AuNPs) were purchased from Nanopartz. They were conjugated with thiolated oligonucleotides specific for the femA gene for the identification of members of Staphylococcus aureus, the mecA gene for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus, hly gene encoding the pore-forming cytolysin listeriolysin for the identification of Listeria monocytogenes and the invA sequence for the identification of Salmonella enteritis. DNA isolation from Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis cultures was performed using the commercial kit Nucleospin Tissue (Macherey Nagel). Specifically 20μl of DNA was diluted in 10mMPBS (pH5). After the denaturation of 10min, 20μl of AuNPs was added followed by the annealing step at 58oC. The presence of a complementary target prevents aggregation with the addition of acid and the solution remains pink, whereas in the opposite event it turns to purple. The color could be detected visually and it was confirmed with an absorption spectrum. Results: Specifically, 0.123 μg/μl DNA of St. aureus, L.monocytogenes and Salmonella enteritis was serially diluted from 1:10 to 1:100. Blanks containing PBS buffer instead of DNA were used. The application of the proposed method on isolated bacteria produced positive results with all the species of St. aureus and L. monocytogenes and Salmonella enteritis using the femA, mecA, hly and invA genes respectively. The minimum detection limit of the assay was defined at 0.2 ng/μL of DNA. Below of 0.2 ng/μL of bacterial DNA the solution turned purple after addition of HCl, defining the minimum detection limit of the assay. None of the blank samples was positive. The specificity was 100%. The application of the proposed method produced exactly the same results every time (n = 4) the evaluation was repeated (100% repeatability) using the femA, hly and invA genes. Using the gene mecA for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus the method had a repeatability 50%. Conclusion: The proposed method could be used as a highly specific and sensitive screening tool for the detection and differentiation of Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis. The use AuNPs for the colorimetric detection of DNA targets represents an inexpensive and easy-to-perform alternative to common molecular assays. The technology described here, may develop into a platform that could accommodate detection of many bacterial species.

Keywords: gold nanoparticles, pathogens, nanotechnology, bacteria

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Authors: Mansoureh Zangiabadi, Shamil Velji, Rajendra Kelkar, Neal Wade, Volker Pickert

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The EU has set itself a long-term goal of reducing greenhouse gas emissions by 80-95% of the 1990 levels by 2050 as set in the Energy Roadmap 2050. This paper reports on the European Union H2020 funded E-Lobster project which demonstrates tools and technologies, software and hardware in integrating the grid distribution, and the railway power systems with power electronics technologies (Smart Soft Open Point - sSOP) and local energy storage. In this context this paper describes the existing policies and regulatory frameworks of the energy market at European level with a special focus then at National level, on the countries where the members of the consortium are located, and where the demonstration activities will be implemented. By taking into account the disciplinary approach of E-Lobster, the main policy areas investigated includes electricity, energy market, energy efficiency, transport and smart cities. Energy storage will play a key role in enabling the EU to develop a low-carbon electricity system. In recent years, Energy Storage System (ESSs) are gaining importance due to emerging applications, especially electrification of the transportation sector and grid integration of volatile renewables. The need for storage systems led to ESS technologies performance improvements and significant price decline. This allows for opening a new market where ESSs can be a reliable and economical solution. One such emerging market for ESS is R+G management which will be investigated and demonstrated within E-Lobster project. The surplus of energy in one type of power system (e.g., due to metro braking) might be directly transferred to the other power system (or vice versa). However, it would usually happen at unfavourable instances when the recipient does not need additional power. Thus, the role of ESS is to enhance advantages coming from interconnection of the railway power systems and distribution grids by offering additional energy buffer. Consequently, the surplus/deficit of energy in, e.g. railway power systems, is not to be immediately transferred to/from the distribution grid but it could be stored and used when it is really needed. This will assure better energy management exchange between the railway power systems and distribution grids and lead to more efficient loss reduction. In this framework, to identify the existing policies and regulatory frameworks is crucial for the project activities and for the future development of business models for the E-Lobster solutions. The projections carried out by the European Commission, the Member States and stakeholders and their analysis indicated some trends, challenges, opportunities and structural changes needed to design the policy measures to provide the appropriate framework for investors. This study will be used as reference for the discussion in the envisaged workshops with stakeholders (DSOs and Transport Managers) in the E-Lobster project.

Keywords: light railway, electrical distribution network, Electrical Energy Storage, policy

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Authors: Nonita Sarin, K.J.Singh, Anuj Kumar, Davinder Singh

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Polymeric 3D hydrogels have pivotal role in bone tissue regeneration applications. Hydrogels behave similar to the living tissues because they have large water imbibing capacity in swollen state and adjust their shape according to the tissues during tissue formation after implantation. On the other hand, hydrogels are very soft, fragile and lack mechanical strength. Incorporation of bioceramics can improve mechanical strength. Furthermore, bioceramics synthesized by sol gel technique may enhance the apatite formation and degradation rates which can lead to the increase in faster rates for new bone and tissue regeneration. Simulated body fluid (SBF) induces the poly-condensation of silanol groups which leads to formation of silica matrix and provide active sites for the precipitation of Ca2+ and PO43- ions to form apatite layer which is similar to mineral form of bone. Therefore, authors have synthesized bioceramic incorporated Polyacrylamide-carboxymethylcellulose hydrogels by free radical polymerization and bioceramic compositions of xSrO-(36-x)CaO-45SiO2-ySeO3-(12-y)P2O5-7MgO (where x=0,4 and y=0,2 mol%) were synthesized by sol gel technique. Bioceramics incorporated in polymer matrix induces quicker apatite formation during immersion in SBF by raising the pH with the release of alkaline ions during ion exchange process and the apatite formation takes place in alkaline medium. The behavior of samples PABC-0 (without bioceramics) and PABC-20 (with 20 wt% bioceramics) were evaluated by X-Ray Diffraction and FTIR. In term of bioactivity, it was observed that PABC-20 has shown hydroxyapatite (HA) formation on 1st day of immersion whereas, PABC-0 was shown apatite formation on 7th day of immersion in SBF. The rapid rate of HA growth on 1st day of immersion in SBF signifies easy regeneration of damaged bone tissues. Degradation studies have been undertaken in Phosphate Buffer Saline and PABC-20 exhibited slower degradation rate up to 9%as compared to PABC-0 up to 18%. Slower degradation rate is suitable for new tissue regeneration and cell attachment. Also, Zeta potential studies have been employed to check the surface charge and it has been observed that samples carry negative charge when immersed in SBF. In addition, the swelling test of the samples have been performed and relative swelling ratio % observed for PABC-0 is 607% and PABC-20 is 305%. This indicates that the incorporation of bioceramics leads to the filling up of the voids in between the polymer matrix which in result reduces porosity and increase the mechanical strength by filling the voids. The porosity of PABC-0 is 84% and PABC-20 is 72%. PABC-20 sample demonstrates that bioceramics incorporation reduce the porosity and improves mechanical strength. Also, maximum in vitro cell viability up to 98% with MG63 cell line has been observed which indicate that the bioceramic incorporated hydrogel(PABC-20) provide the alkaline medium which is suitable environment for cell growth.

Keywords: hydrogels, hydroxyapatite, MG63 cell line, zeta potential

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Authors: Hollie Ashworth, Sarah Heath, Nick Bryan, Liam Abrahamsen, Simon Kellet

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Sellafield has a number of fuel storage ponds, some of which have been open to the air for a number of decades. This has caused corrosion of the fuel resulting in a release of some activity into solution, reduced water clarity, and accumulation of sludge at the bottom of the pond consisting of brucite (Mg(OH)2) and other uranium corrosion products. Both of these phases are also present as colloidal material. 90Sr and 137Cs are known to constitute a small volume of the radionuclides present in the pond, but a large fraction of the activity, thus they are most at risk of challenging effluent discharge limits. Organic molecules are known to be present also, due to the ponds being open to the air, with occasional algal blooms restricting visibility further. The contents of the pond need to be retrieved and safely stored, but dealing with such a complex, undefined inventory poses a unique challenge. This work aims to determine and understand the sorption-desorption interactions of 90Sr and 137Cs to brucite and uranium phases, with and without the presence of organic molecules from chemical degradation and bio-organisms. The influence of organics on these interactions has not been widely studied. Partitioning of these radionuclides and organic molecules has been determined through LSC, ICP-AES/MS, and UV-vis spectrophotometry coupled with ultrafiltration in both binary and ternary systems. Further detailed analysis into the surface and bonding environment of these components is being investigated through XAS techniques and PHREEQC modelling. Experiments were conducted in CO2-free or N2 atmosphere across a high pH range in order to best simulate conditions in the pond. Humic acid used in brucite systems demonstrated strong competition against 90Sr for the brucite surface regardless of the order of addition of components. Variance of pH did have a small effect, however this range (10.5-11.5) is close to the pHpzc of brucite, causing the surface to buffer the solution pH towards that value over the course of the experiment. Sorption of 90Sr to UO2 obeyed Ho’s rate equation and demonstrated a slow second-order reaction with respect to the sharing of valence electrons from the strontium atom, with the initial rate clearly dependent on pH, with the equilibrium concentration calculated at close to 100% sorption. There was no influence of humic acid seen when introduced to these systems. Sorption of 137Cs to UO3 was significant, with more than 95% sorbed in just over 24 hours. Again, humic acid showed no influence when introduced into this system. Both brucite and uranium based systems will be studied with the incorporation of cyanobacterial cultures harvested at different stages of growth. Investigation of these systems provides insight into, and understanding of, the effect of organics on radionuclide partitioning to brucite and uranium phases at high pH. The majority of sorption-desorption work for radionuclides has been conducted at neutral to acidic pH values, and mostly without organics. These studies are particularly important for the characterisation of legacy wastes at Sellafield, with a view to their safe retrieval and storage.

Keywords: caesium, legacy wastes, organics, sorption-desorption, strontium, uranium

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