Search results for: biological affection

Authors: Paula K. Novelli, Margarida M. Barros, Luciana F. Flueri

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Utilization of agricultural by-products, wheat ban and soybean bran, as substrate for solid state fermentation (SSF) was studied, aiming the achievement of different enzymes from Aspergillus sp. with distinct biological characteristics and its application and improvement on animal nutrition. Aspergillus niger and Aspergillus oryzea were studied as they showed very high yield of phytase and protease production, respectively. Phytase activity was measure using p-nitrophenilphosphate as substrate and a standard curve of p-nitrophenol, as the enzymatic activity unit was the quantity of enzyme necessary to release one μmol of p-nitrophenol. Protease activity was measure using azocasein as substrate. Activity for phytase and protease substantially increased when the different biochemical characteristics were considered in the study. Optimum pH and stability of the phytase produced by A. niger with wheat bran as substrate was between 4.0 - 5.0 and optimum temperature of activity was 37oC. Phytase fermented in soybean bran showed constant values at all pHs studied, for optimal and stability, but low production. Phytase with both substrates showed stable activity for temperatures higher than 80oC. Protease from A. niger showed very distinct behavior of optimum pH, acid for wheat bran and basic for soybean bran, respectively and optimal values of temperature and stability at 50oC. Phytase produced by A. oryzae in wheat bran had optimum pH and temperature of 9 and 37oC, respectively, but it was very unstable. On the other hand, proteases were stable at high temperatures, all pH’s studied and showed very high yield when fermented in wheat bran, however when it was fermented in soybean bran the production was very low. Subsequently the upscale production of phytase from A. niger and proteases from A. oryzae were applied as an enzyme additive in fish fed for digestibility studies. Phytases and proteases were produced with stable enzyme activity of 7,000 U.g-1 and 2,500 U.g-1, respectively. When those enzymes were applied in a plant protein based fish diet for digestibility studies, they increased protein, mineral, energy and lipids availability, showing that these new enzymes can improve animal production and performance. In conclusion, the substrate, as well as, the microorganism species can affect the biochemical character of the enzyme produced. Moreover, the production of these enzymes by SSF can be up to 90% cheaper than commercial ones produced with the same fungi species but submerged fermentation. Add to that these cheap enzymes can be easily applied as animal diet additives to improve production and performance.

Keywords: agricultural by-products, animal nutrition, enzymes production, solid state fermentation

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Authors: Davendra S. Malik

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Bhimtal lake is located between 290 21’ N latitude and 790 24’ E longitude, at an elevation of 1332m above mean sea level in the Kumaun region of Uttarakhand of Indian subcontinent. The lake surface area is decreasing in water area, depth level in relation to ecological and biological characteristics due to climatic variations, invasive land use pattern, degraded forest zones and changed agriculture pattern in lake catchment basin. The present study is focused on long and short term effects of climate change on aquatic biodiversity and productivity of Bhimtal lake. The meteorological data of last fifteen years of Bhimtal lake catchment basin revealed that air temperature has been increased 1.5 to 2.1oC in summer, 0.2 to 0.8 C in winter, relative humidity increased 4 to 6% in summer and rainfall pattern changed erratically in rainy seasons. The surface water temperature of Bhimtal lake showed an increasing pattern as 0.8 to 2.6 C, pH value decreased 0.5 to 0.2 in winter and increased 0.4 to 0.6 in summer. Dissolved oxygen level in lake showed a decreasing trend as 0.7 to 0.4mg/l in winter months. The mesotrophic nature of Bhimtal lake is changing towards eutrophic conditions and contributed for decreasing biodiversity. The aquatic biodiversity of Bhimtal lake consisted mainly phytoplankton, zooplankton, benthos and fish species. In the present study, a total of 5 groups of phytoplankton, 3 groups of zooplankton, 11 groups of benthos and 15 fish species were recorded from Bhimtal lake. The comparative data of biodiversity of Bhimtal lake since January, 2000 indicated the changing pattern of phytoplankton biomass were decreasing as 1.99 and 1.08% of Chlorophyceae and Bacilleriophyceae families respectively. The biomass of Cynophyceae was increasing as 0.45% and contributing the algal blooms during summer season in lake. The biomass of zooplankton and benthos were found decreasing in winter season and increasing during summer season. The endemic fish species (18 no.) were found in year 2000-05, as while the fish species (15 no.) were recorded in present study. The relative fecundity of major fish species were observed decreasing trends during their breeding periods in lake. The natural and anthropogenic factors were identified as ecological threats for existing aquatic biodiversity of Bhimtal lake. The present research paper emphasized on the effect of changing pattern of different climatic variables on species composition, biomass of phytoplankton, zooplankton, benthos, and fishes in Bhimtal lake of Kumaun region. The present research data will be contributed significantly to assess the changing pattern of aquatic biodiversity and productivity of Bhimtal lake with different time scale.

Keywords: aquatic biodiversity, Bhimtal lake, climate change, lake ecology

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Authors: Maria David, Tunde A. Tasko, Katalin Hejja-Nagy, Laszlo Dorner

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The authors have been concerned with research on learning since 1998. Recently, the focus of our interest is how prevalent use of information and communication technology (ICT) influences students' learning abilities, skills of self-regulated learning and learning efficiency. Nowadays, there are three dominant theories about the psychic effects of ICT use: According to social optimists, modern ICT devices have a positive effect on thinking. As to social pessimists, this effect is rather negative. And, regarding the views of biological optimists, the change is obvious, but these changes can fit into the mankind's evolved neurological system as did writing long ago. Mentality of 'digital natives' differ from that of elder people. They process information coming from the outside world in an other way, and different experiences result in different cerebral conformation. In this regard, researchers report about both positive and negative effects of ICT use. According to several studies, it has a positive effect on cognitive skills, intelligence, school efficiency, development of self-regulated learning, and self-esteem regarding learning. It is also proven, that computers improve skills of visual intelligence such as spacial orientation, iconic skills and visual attention. Among negative effects of frequent ICT use, researchers mention the decrease of critical thinking, as permanent flow of information does not give scope for deeper cognitive processing. Aims of our present study were to uncover developmental characteristics of self-regulated learning in different age groups and to study correlations of learning efficiency, the level of self-regulated learning and frequency of use of computers. Our subjects (N=1600) were primary and secondary school students and university students. We studied four age groups (age 10, 14, 18, 22), 400 subjects of each. We used the following methods: the research team developed a questionnaire for measuring level of self-regulated learning and a questionnaire for measuring ICT use, and we used documentary analysis to gain information about grade point average (GPA) and results of competence-measures. Finally, we used computer tasks to measure cognitive abilities. Data is currently under analysis, but as to our preliminary results, frequent use of computers results in shorter response time regarding every age groups. Our results show that an ordinary extent of ICT use tend to increase reading competence, and had a positive effect on students' abilities, though it didn't show relationship with school marks (GPA). As time passes, GPA gets worse along with the learning material getting more and more difficult. This phenomenon draws attention to the fact that students are unable to switch from guided to independent learning, so it is important to consciously develop skills of self-regulated learning.

Keywords: digital natives, ICT, learning efficiency, reading competence, self-regulated learning

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Authors: I. Cerrillo, A. Carrillo-Vico, M. A. Ortega, B. Escudero-López, N. Álvarez-Sánchez, F. Martín, M. S. Fernández-Pachón

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Melatonin is a bioactive compound involved in multiple biological activities such as glucose tolerance, circadian rhythm regulation, antioxidant defense or immune system action. In elderly subjects the intake of foods and drinks rich in melatonin is very important due to its endogenous level decreases with age. Alcoholic fermentation is a process carried out in fruits, vegetables and legumes to obtain new products with improved bioactive compounds profile in relation to original substrates. Alcoholic fermentation process carried out by Saccharomycetaceae var. Pichia kluyveri induces an important synthesis of melatonin in orange juice. A novel beverage derived of fermented orange juice could be a promising source of this bioactive compound. The aim of the present study was to determine whether the acute intake of fermented orange juice increase the levels of urinary 6-sulfatoxymelatonin in healthy humans. Nine healthy volunteers (7 women and 2 men), aged between 20 and 25 years old and BMI of 21.1  2.4 kg/m2, were recruited. On the study day, participants ingested 500 mL of fermented orange juice. The first urine collection was made before fermented orange juice consumption (basal). The rest of urine collections were made in the following time intervals after fermented orange juice consumption: 0-2, 2-5, 5-10, 10- 15 and 15-24 hours. During the experimental period only the consumption of water was allowed. At lunch time a meal was provided (60 g of white bread, two slices of ham, a slice of cheese, 125 g of sweetened natural yoghurt and water). The subjects repeated the protocol with orange juice following a 2-wk washout period between both types of beverages. The levels of 6-sulfatoxymelatonin (6-SMT) were measured in urine recollected at different time points using the Melatonin-Sulfate Urine ELISA (IBL International GMBH, Hamburg, Germany). Levels of 6-SMT were corrected to those of creatinine for each sample. A significant (p < 0.05) increase in urinary 6-SMT levels was observed between 2-5 hours after fermented orange juice ingestion with respect to basal values (increase of 67,8 %). The consumption of orange juice did not induce any significant change in urinary 6-SMT levels. In addition, urinary 6-SMT levels obtained between 2-5 hours after fermented orange juice ingestion (115,6 ng/mg) were significantly different (p < 0.05) from those of orange juice (42,4 ng/mg). The enhancement of urinary 6-SMT after the ingestion of 500 mL of fermented orange juice in healthy humans compared to orange juice could be an important advantage of this novel product as an excellent source of melatonin. Fermented orange juice could be a new functional food, and its consumption could exert a potentially positive effect on health in both the maintenance of health status and the prevention of chronic diseases.

Keywords: fermented orange juice, functional beverage, healthy human, melatonin

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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: Gergely Horvath

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The study analyzes the special provisions of the highest level of national agricultural legislation in the Fundamental Law of Hungary (25 April 2011) with descriptive, analytic and comparative methods. The agriculturally relevant articles of the constitution are very important, because –in spite of their high level of abstraction– they can determine and serve the practice comprehensively and effectively. That is why the objective of the research is to interpret the concrete sentences and phrases in connection with agriculture compared with the methods of some other relevant constitutions (historical-grammatical interpretation). The major findings of the study focus on searching for the appropriate provisions and approach capable of solving the problems of sustainable food production. The real challenge agricultural law must face with in the future is protecting or conserving its background and subjects: the environment, the ecosystem services and all the 'roots' of food production. In effect, agricultural law is the legal aspect of the production of 'our daily bread' from farm to table. However, it also must guarantee the safe daily food for our children and for all our descendants. In connection with sustainability, this unique, value-oriented constitution of an agrarian country even deals with uncustomary questions in this level of legislation like GMOs (by banning the production of genetically modified crops). The starting point is that the principle of public good (principium boni communis) must be the leading notion of the norm, which is an idea partly outside the law. The public interest is reflected by the agricultural law mainly in the concept of public health (in connection with food security) and the security of supply with healthy food. The construed Article P claims the general protection of our natural resources as a requirement. The enumeration of the specific natural resources 'which all form part of the common national heritage' also means the conservation of the grounds of sustainable agriculture. The reference of the arable land represents the subfield of law of the protection of land (and soil conservation), that of the water resources represents the subfield of water protection, the reference of forests and the biological diversity visualize the specialty of nature conservation, which is an essential support for agrobiodiversity. The mentioned protected objects constituting the nation's common heritage metonymically melt with their protective regimes, strengthening them and forming constitutional references of law. This regimes also mean the protection of the natural foundations of the life of the living and also the future generations, in the name of intra- and intergenerational equity.

Keywords: agricultural law, constitutional values, natural resources, sustainability

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Authors: Kailash Chandra, Devanshu Gupta

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The Himalayan Mountains of India fall under two biogeographic zones Trans Himalaya (TH) and Himalaya and seven biotic provinces (TH-Ladakh Mountains, TH-Tibetan Plateau, TH-Sikkim, North-West Himalaya, West Himalaya, Central Himalaya, and East Himalaya). Because of the extreme environment and altitudinal variations, unique physiography, varied ecological conditions, and different vegetations, the Himalaya exhibit a rich assemblage of life, both flora, and fauna, further subjected to the impacts of climate change. To the authors’ best knowledge, there is no comprehensive account except for sporadic faunal investigations, to assess or interpret the insect diversity and their biogeographic distribution in Indian Himalaya (IH), one of the biodiversity hotspots. Therefore, in this paper, a compelling review of the extensive knowledge of insect diversity of IH is presented for the first time to the best of our knowledge. The inventory of the known insect species of IH was compiled from the exploration cum faunal-study data ready with the zoological survey of India, Kolkata as well as from the information published in the scientific literature till date. The species were listed with their valid names with their distribution in seven biotic provinces of IH. The insect fauna of IH represents about 38% of the identified insect diversity of India. The interpretation of data provided significant information in detecting possible gap areas in the taxonomic representation of different insect orders. Archaeognatha, Zygentoma, Ephemeroptera, Phasmida, Embioptera, Psocoptera, Phthiraptera, Strepsiptera, Megaloptera, Raphidioptera, Siphonaptera, and Mecoptera need revisions, and it is required to collect more samples from remote areas of the region. Scope for finding new taxa even in the most diverse orders, Coleoptera, Lepidoptera, Hymenoptera, Diptera, and Hemiptera cannot be overlooked. Exploration of cold deserts of Trans Himalaya and East Himalaya (Arunachal Pradesh) may result in a good number of new species from these regions. The most notable data was that many of the species recorded from Himalaya are still known from their type localities only, so there is an urgency to revisit and resurvey those collection localities for the evaluation of the status of those species. It is also required to assess and monitor the impact of climate change on the diversity of insects inhabiting in the fragile Himalayan ecosystem. DNA barcoding especially pests and biological control agents to solve the problems of identification in species complexes is also the need of the hour. In a nutshell, it can be concluded that the inventory of insects of this region is extensive but is far from final as every year hundreds of new species are described.

Keywords: catalog, climate change, diversity, DNA barcoding

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Authors: Jacob Lewis Watts, Adam F. A. Pellegrini

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Prairies, grasslands, and forests cover an expansive portion of the world’s surface and contribute significantly to Earth’s carbon cycle. The largest driver of carbon dynamics in some of these ecosystems is fire. As the global climate changes, most fire-dominated ecosystems will experience increased fire frequency and intensity, leading to increased carbon flux into the atmosphere and soil nutrient depletion. The plant communities associated with different fire regimes are important for reassimilation of carbon lost during fire and soil recovery. More frequent fires promote conservative plant functional traits aboveground; however, belowground fine root traits are poorly explored and arguably more important drivers of ecosystem function as the primary interface between the soil and plant. The root economic spectrum (RES) hypothesis describes single-dimensional covariation between important fine-root traits along a range of plant strategies from acquisitive to conservative – parallel to the well-established leaf economic spectrum (LES). However, because of the paucity of root trait data, the complex nature of the rhizosphere, and the phylogenetic conservatism of root traits, it is unknown whether the RES hypothesis accurately describes plant nutrient and water acquisition strategies. This project utilizesplants grown in common garden conditions in the Cambridge University Botanic Garden and a meta-analysis of long-term fire manipulation experiments to examine the belowground physiological traits of fire-adapted and non-fire-adapted herbaceous species to 1) test the RES hypothesis and 2) describe the effect of fire regimes on fine root functional traits – which in turn affect carbon and nutrient cycling. A suite of morphological, chemical, and biological root traits (e.g. root diameter, specific root length, percent N, percent mycorrhizal colonization, etc.) of 50 herbaceous species were measuredand tested for phylogenetic conservatism and RES dimensionality. Fire-adapted and non-fire-adapted plants traits were compared using phylogenetic PCA techniques. Preliminary evidence suggests that phylogenetic conservatism may weaken the single-dimensionality of the RES, suggesting that there may not be a single way that plants optimize nutrient and water acquisition and storage in the complex rhizosphere; additionally, fire-adapted species are expected to be more conservative than non-fire-adapted species, which may be indicative of slower carbon cycling with increasing fire frequency and intensity.

Keywords: climate change, fire regimes, root economic spectrum, fine roots

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Authors: Sofia Lazareva, Artem Smolentsev

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Fluorescent photochromic materials collect strong interest due to their possible application in organic photonics such as optical logic systems, optical memory, visualizing sensors, as well as characterization of polymers and biological systems. In photochromic fluorescence switching systems the emission of fluorophore is modulated between ‘on’ and ‘off’ via the photoisomerization of photochromic moieties resulting in effective resonance energy transfer (FRET). In current work, we have studied both photochromic and fluorescent properties of several diarylethenes. It was found that coloured forms of these compounds are not fluorescent because of the efficient intramolecular energy transfer. Spectral and photochromic parameters of investigated substances have been measured in five solvents having different polarity. Quantum yields of photochromic transformation A↔B ΦA→B and ΦB→A as well as B isomer extinction coefficients were determined by kinetic method. It was found that the photocyclization reaction quantum yield of all compounds decreases with the increase of solvent polarity. In addition, the solvent polarity is revealed to affect fluorescence significantly. Increasing of the solvent dielectric constant was found to result in a strong shift of emission band position from 450 nm (nhexane) to 550 nm (DMSO and ethanol) for all three compounds. Moreover, the emission intensive in polar solvents becomes weak and hardly detectable in n-hexane. The only one exception in the described dependence is abnormally low fluorescence quantum yield in ethanol presumably caused by the loss of electron-donating properties of nitrogen atom due to the protonation. An effect of the protonation was also confirmed by the addition of concentrated HCl in solution resulting in a complete disappearance of the fluorescent band. Excited state dynamics were investigated by ultrafast optical spectroscopy methods. Kinetic curves of excited states absorption and fluorescence decays were measured. Lifetimes of transient states were calculated from the data measured. The mechanism of ring opening reaction was found to be polarity dependent. Comparative analysis of kinetics measured in acetonitrile and hexane reveals differences in relaxation dynamics after the laser pulse. The most important fact is the presence of two decay processes in acetonitrile, whereas only one is present in hexane. This fact supports an assumption made on the basis of steady-state preliminary experiments that in polar solvents occur stabilization of TICT state. Thus, results achieved prove the hypothesis of two channel mechanism of energy relaxation of compounds studied.

Keywords: diarylethenes, fluorescence switching, FRET, photochromism, TICT state

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Authors: A. Kesraoui, M. Seffen

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Applications of adsorption onto activated carbon for water treatment are well known. The process has been demonstrated to be widely effective for removing dissolved organic substances from wastewaters, but this treatment has a major drawback is the high operating cost. The main goal of our research work is to improve the retention capacity of Tunisian biomass for the depollution of industrial wastewater and retention of pollutants considered toxic. The biosorption process is based on the retention of molecules and ions onto a solid surface composed of biological materials. The evaluation of the potential use of these materials is important to propose as an alternative to the adsorption process generally expensive, used to remove organic compounds. Indeed, these materials are very abundant in nature and are low cost. Certainly, the biosorption process is effective to remove the pollutants, but it presents a slow kinetics. The improvement of the biosorption rates is a challenge to make this process competitive with respect to oxidation and adsorption onto lignocellulosic fibers. In this context, the alternating current appears as a new alternative, original and a very interesting phenomenon in the acceleration of chemical reactions. Our main goal is to increase the retention acceleration of dyes (indigo carmine, methylene blue) and phenol by using a new alternative: alternating current. The adsorption experiments have been performed in a batch reactor by adding some of the adsorbents in 150 mL of pollutants solution with the desired concentration and pH. The electrical part of the mounting comprises a current source which delivers an alternating current voltage of 2 to 15 V. It is connected to a voltmeter that allows us to read the voltage. In a 150 mL capacity cell, we plunged two zinc electrodes and the distance between two Zinc electrodes has been 4 cm. Thanks to alternating current, we have succeeded to improve the performance of activated carbon by increasing the speed of the indigo carmine adsorption process and reducing the treatment time. On the other hand, we have studied the influence of the alternating current on the biosorption rate of methylene blue onto Luffa cylindrica fibers and the hybrid material (Luffa cylindrica-ZnO). The results showed that the alternating current accelerated the biosorption rate of methylene blue onto the Luffa cylindrica and the Luffa cylindrica-ZnO hybrid material and increased the adsorbed amount of methylene blue on both adsorbents. In order to improve the removal of phenol, we performed the coupling between the alternating current and the biosorption onto two adsorbents: Luffa cylindrica and the hybrid material (Luffa cylindrica-ZnO). In fact, the alternating current has succeeded to improve the performance of adsorbents by increasing the speed of the adsorption process and the adsorption capacity and reduce the processing time.

Keywords: adsorption, alternating current, dyes, modeling

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Authors: Muhammad Naeem Chaudhry, Fahim Nawaz, Rana Nauman Shabbir

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New strategies aimed at increasing the resilience of crop plants to the negative effects of climate change represent important research priorities of plant scientists. The use of soil microorganisms to alleviate abiotic stresses like drought has gained particular importance in recent past. A field experiment was planned to investigate the effect of phosphorous solubilizing bacteria on yield and seed quality of Camelina (Camelina sativa L.) under water deficit conditions. The study was conducted at Agronomic Research Farm, University College of Agriculture and Environmental Sciences, The Islamia University Bahawalpur, during 4th week of November, 2013. The available seeds of Camelina sativa were inoculated with two bacterial strains (pseudomonas and Bacillus spp.) and grown under various water stress levels i.e. D0, (four irrigations), D3 (three irrigation), D2 (two irrigations), and D1 (one irrigation). The results revealed that drought stress significantly reduced the plant growth and yield, consequently reducing protein contents and oil concentration in camelina. The exposure to drought stress decreased plant height (16%), plant population (27%), number of fertile branches (41-59%), number of pods per plant (35%) and seed per pod (33%). Drought stress also exerted a negative impact on yield characteristics by reducing the 1000-seed weight (65%), final seed yield (52%), biological yield (22%) and harvest index (39%) of camelina. However, the inoculation of seeds with Pseudomonas and Bacillus spp. promoted the plant growth characterized by increased plant height and enhanced plant population. It was noted that inoculation of seeds with Pseudomonas resulted in the maximum plant population (113.4 cm), primary branches (19 plant-1), and number of pods (664 plant-1), whereas Bacillus inoculation resulted in maximum plant height (113.4 cm), seeds per pod (15.9), 1000-seed weight (1.85 g), and seed yield (3378.8 kg ha-1). Moreover, the inoculation with Bacillus also significantly improved the quality attributes of camelina and gave 3.5% and 2.1% higher oil contents than Pseudomonas and control (no-inoculation), respectively. Similarly, the same strain also resulted in maximum protein contents (33.3%). Our results confirmed the hypothesis that inoculation of seeds with phosphorous solubilizing bacterial strains is an effective, viable and environment-friendly approach to improve yield and quality of camelina under water deficit conditions. However, further studies are suggested to investigate the physiological and molecular processes, stimulated by bacterial strains, for increasing drought tolerance in food crops.

Keywords: Camelina, drought stress, phosphate solubilizing bacteria, seed quality

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Authors: Abdolamir Allameh, Shahnaz Esmaeili, Mina Allameh, Safoura Khajeniazi

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Stem cells with therapeutic applications can be isolated from human placenta/umblical cord blood (UCB) as well as the cord tissue (UC). Stem cells in culture are vulnerable to oxidative stress, particularly when subjected to differentiation process. The aim of this study was to examine the chnages in the rate of oxidation that occurs to cellular macromolecules during hepatic differentiation of mononuclear cells (MSCs). In addition, the impact of the hepatic differentiation process of MSC on cellular and biological activity of the cells will be undertaken. For this purpose, first mononuclear cells (MNCs) were isolated from human UCB which was obtained from a healthy full-term infant. The cells were cultured at a density of 3×10⁵ cells/cm² in DMEM- low-glucose culture media supplemented with 20% FBS, 2 mM L-glutamine, 100 μg/ml streptomycin and 100 U/ml penicillin. Cell cultures were then incubated at 37°C in a humidified 5% CO₂ incubator. After removing non-adherent cells by replacing culture medium, fibroblast-like adherent cells were resuspended in 0.25% trypsin-EDTA and plated in 25 cm² flasks (1×10⁴/ml). Characterization of the MSCs was routinely done by observing their morphology and growth curve. MSCs were subjected to a 2-step hepatocyte differentiation protocol in presence of hepatocyte growth factor (HGF), dexamethazone (DEX) and oncostatin M (OSM). The hepatocyte-like cells derived from MSCs were checked every week for 3 weeks for changes in lipid peroxidation, protein carbonyl formation and DNA oxidation i.e., 8-hydroxy-2'-deoxyguanosine (8-OH-dG) assay. During the 3-week differentiation process of MSCs to hepatocyte-like cells we found that expression liver-specific markers such as albumin, was associated with increased levels of lipid peroxidation and protein carbonyl formation. Whereas, undifferentiated MSCs has relatively low levels of lipid peroxidation products. There was a significant increase ( p < 0.05) in lipid peroxidation products in hepatocytes on days 7, 14, and 21 of differentiation. Likewise, the level of protein carbonyls in the cells was elevated during the differentiation. The level of protein carbonyls measured in hepatocyte-like cells obtained 3 weeks after differentiation induction was estimated to be ~6 fold higher compared to cells recovered on day 7 of differentiation. On the contrary, there was a small but significant decrease in DNA damage marker (8-OH-dG) in hepatocytes recovered 3 weeks after differentiation onset. The level of 8-OHdG which was in consistent with formation of reactive oxygen species (ROS). In conclusion, this data suggest that despite the elevation in oxidation of lipid and protein molecules during hepatocyte development, the cells were normal in terms of DNA integrity, morphology, and biologically activity.

Keywords: adult stem cells, DNA integrity, free radicals, hepatic differentiation

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Authors: E. Rommes, Els Toonen, Rahil Roodsaz, Suborna Camellia, Farhana Alam, Saad Khan, Jhalok Ranjon Talukder, Tanveer Hassan, Syeda Farjana Ahmed, Sabina Faiz Rashid

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In Bangladesh, a country in which adults (both parents and teachers) find it particularly hard to speak with youth about sexuality, adolescents seem to struggle with various insecurities about their sexual feelings, thoughts, behavior and physical characteristics. On the basis of a large number of interviews and focus groups with rural and urban Bangla adolescent girls and boys of lower and middle class as part of the large-scale three-year project ‘Breaking the Shame’, we have identified ten sexual themes or ‘myths’ that youth struggle with most. These encompass amongst others beliefs and insecurities on masturbation, discharge, same-sex behavior and feelings, the effects of watching porn and gender norms. We argue that the Internet is a particularly suitable medium to ‘debunk’ those myths, as youth can consult it anonymously and privately and so avoid social shame. Moreover, amongst the myths, we have identified two kinds which may need different debunking techniques. One kind of myth concerns scientifically uncontested, generally biological related information, such as the effects of having sex with a pregnant woman, questions on the effects of a penile or vaginal discharge or questions on the effects of masturbation. The second kind of myths concerns more diverse information sources and deals with e.g. religious or culturally specific norms, such as on the meaning and existence of homosexuality or gender appropriate norms of behavior in Bangladesh. For addressing both kinds of myths, expert information including a wealth of references to information resources needs to be provided, which the Internet is very suitable for. For the second kind of myths, adolescents also need to learn how to deal with sometimes conflicting norms and information sources, and they need to develop and reflect on their own opinions as part of their identity formation. On the basis of a literature review, we thus distinguish general information needs from identity formation needs, which includes the need to be able to relate information and opinions to one’s own opinions and situation. Hence, we argue that youth not only need abstract expert information to be able to debunk sexual myths, but also the option to discuss this information with other adolescents and compare their own situation and opinions with other peers, who in that way serve as ‘warm experts’ for each other. In this paper, we will describe the outcomes of our qualitative study above. In addition, we will present our findings of an intervention by presenting youth with general, uncontested information on the Internet with additional peer discussion options to compare the debunking effects on different kinds of myths.

Keywords: peer discussion, intervention, sexual myths, shame

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Authors: N. Selvi Gunel, L. M. Oktay, H. Memmedov, B. Durmaz, H. Kalkan Yildirim, E. Yildirim Sozmen

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Object: Propolis which consists of compounds that are accepted as antioxidant, antimicrobial, antiseptic, antibacterial, anti-inflammatory, anti-mutagenic, immune-modulator and cytotoxic, is frequently used in current therapeutic applications. However, some of them result in allergic side effects, causing consumption to be restricted. Previously our group has succeeded in producing a new biotechnological product which was less allergenic. In this study, we purpose to optimize production conditions of this biologically-transformed propolis and determine the cytotoxic effects of obtained new products on colon cancer cell line (HCT-116). Method: Firstly, solid propolis samples were dissolved in water after weighing, grinding and sizing (sieve-35mesh) and applied 40 kHz/10 min ultrasonication. Samples were prepared according to inoculation with Lactobacillus plantarum in two different proportions (2.5% and 3.5%). Chromatographic analyzes of propolis were performed by UPLC-MS/MS (Waters, Milford, MA) system. Results were analysed by UPLC-MS/MS system MassLynx™ 4.1 software. HCT-116 cells were treated with propolis examples at 25-1000 µg/ml concentrations and cytotoxicity were measured by using WST-8 assay at 24, 48, and 72 hours. Samples with biological transformation were compared with the non-transformed control group samples. Our experiment groups were formed as follows: untreated (group 1), propolis dissolved in water ultrasonicated at 40 kHz/10 min (group 2), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 2.5% L. plantarum L1 strain (group 3), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 3.5% L. plantarum L3 strain (group 4). Obtained data were calculated with Graphpad Software V5 and analyzed by two-way ANOVA test followed by Bonferroni test. Result: As a result of our study, the cytotoxic effect of propolis samples on HCT-116 cells was evaluated. There was a 7.21 fold increase in group 3 compared to group 2 in the concentration of 1000 µg/ml, and it was a 6.66 fold increase in group 3 compared to group 1 at the end of 24 hours. At the end of 48 hours, in the concentration of 500 µg/ml, it was determined 4.7 fold increase in group 4 compared to group 3. At the same time, in the concentration of 750 µg/ml it was determined 2.01 fold increase in group 4 compared to group 3 and in the same concentration, it was determined 3.1 fold increase in group 4 compared to group 2. Also, at the 72 hours, in the concentration of 750 µg/ml, it was determined 2.42 fold increase in group 3 according to group 2 and in the same time, in the concentration of 1000 µg/ml, it was determined 2.13 fold increase in group 4 according to group 2. According to cytotoxicity results, the group which were ultrasonicated at 40 kHz/10min and inoculated 3.5% L. plantarum L3-strain had a higher cytotoxic effect. Conclusion: It is known that bioavailability of propolis is halved in six months. The data obtained from our results indicated that biologically-transformed propolis had more cytotoxic effect than non-transformed group on colon cancer cells. Consequently, we suggested that L. plantarum-transformation provides both reduction of allergenicity and extension of bioavailability period by enhancing healthful polyphenols.

Keywords: bio-transformation, propolis, colon cancer, cytotoxicity

Procedia PDF Downloads 140

Authors: Mmatladi D. Mnguni, Mustapha Mohammed, George Y. Mahama, Alhassan L. Abdulai, Felix D. Dakora

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Industrial-based nitrogen (N) fertilizers are justifiably credited for the current state of food production across the globe, but their continued use is not sustainable and has an adverse effect on the environment. The search for greener and sustainable technologies has led to an increase in exploiting biological systems such as legumes and organic amendments for plant growth promotion in cropping systems. Although the benefits of legume rotation with cereal crops have been documented, the full benefits of soybean-sorghum rotation systems have not been properly evaluated in Africa. This study explored the benefits of soybean-sorghum rotation through assessing N₂ fixation and water-use efficiency of soybean in rotation with sorghum with and without organic and inorganic amendments. The field trials were conducted from 2017 to 2020. Sorghum was grown on plots previously cultivated to soybean and vice versa. The succeeding sorghum crop received fertilizer amendments [organic fertilizer (5 tons/ha as poultry litter, OF); inorganic fertilizer (80N-60P-60K) IF; organic + inorganic fertilizer (OF+IF); half organic + inorganic fertilizer (HIF+OF); organic + half inorganic fertilizer (OF+HIF); half organic + half inorganic (HOF+HIF) and control] and was arranged in a randomized complete block design. The soybean crop succeeding fertilized sorghum received a blanket application of triple superphosphate at 26 kg P ha⁻¹. Nitrogen fixation and water-use efficiency were respectively assessed at the flowering stage using the ¹⁵N and ¹³C natural abundance techniques. The results showed that the shoot dry matter of soybean plants supplied with HOF+HIF was much higher (43.20 g plant-1), followed by OF+HIF (36.45 g plant⁻¹), and HOF+IF (33.50 g plant⁻¹). Shoot N concentration ranged from 1.60 to 1.66%, and total N content from 339 to 691 mg N plant⁻¹. The δ¹⁵N values of soybean shoots ranged from -1.17‰ to -0.64‰, with plants growing on plots previously treated to HOF+HIF exhibiting much higher δ¹⁵N values, and hence lower percent N derived from N₂ fixation (%Ndfa). Shoot %Ndfa values varied from 70 to 82%. The high %Ndfa values obtained in this study suggest that the previous year’s organic and inorganic fertilizer amendments to sorghum did not inhibit N₂ fixation in the following soybean crop. The amount of N-fixed by soybean ranged from 106 to 197 kg N ha⁻¹. The treatments showed marked variations in carbon (C) content, with HOF+HIF treatment recording the highest C content. Although water-use efficiency varied from -29.32‰ to -27.85‰, shoot water-use efficiency, C concentration, and C:N ratio were not altered by previous fertilizer application to sorghum. This study provides strong evidence that previous HOF+HIF sorghum residues can enhance N nutrition and water-use efficiency in nodulated soybean.

Keywords: ¹³C and ¹⁵N natural abundance, N-fixed, organic and inorganic fertilizer amendments, shoot %Ndfa

Procedia PDF Downloads 170

Authors: V. Stejskal, K. Lundova, R. Sebesta, T. Vanina, S. Roje

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Pure strain of Arctic char (AC) Salvelinus alpinus and hybrid (HB) Salvelinus alpinus vs. Salvelinus fontinalis Mitchill belong to fish, which with great potential for culture in recirculating aquaculture systems (RAS). Aquaculture of these fish currently use flow-through systems (FTS), especially in Nordic countries such as Iceland (biggest producer), Norway, Sweden, and Canada. Four different water saturation regimes included normoxia (NOR), permanent hyperoxia (HYP), intermittent hyperoxia (HYP ± ) and regimes where one day of normoxia was followed by one day of hyperoxia (HYP1/1) were tested during 63 days of experiment in both species in two parallel experiments. Fish were reared in two identical RAS system consisted of 24 plastic round tanks (300 L each), drum filter, biological filter with moving beads and submerged biofilter. The temperature was maintained using flow-through cooler during at level of 13.6 ± 0.8 °C. Different water saturation regimes were achieved by mixing of pure oxygen (O₂) with water in three (one for each hyperoxic regime) mixing tower equipped with flowmeter for regulation of gas inflow. The water in groups HYP, HYP1/1 and HYP± was enriched with oxygen up to saturation of 120-130%. In HYP group was this level kept during whole day. In HYP ± group was hyperoxia kept for daylight phase (08:00-20:00) only and during night time was applied normoxia in this group. The oxygen saturation of 80-90% in NOR group was created using intensive aeration in header tank. The fish were fed with commercial feed to slight excess at 2 h intervals within the light phase of the day. Water quality parameters like pH, temperature and level of oxygen was monitoring three times (7 am, 10 am and 6 pm) per day using handy multimeter. Ammonium, nitrite and nitrate were measured in two day interval using spectrophotometry. Initial body weight (BW) was 40.9 ± 8.7 g and 70.6 ± 14.8 in AC and HB group, respectively. Final survival of AC ranged from 96.3 ± 4.6 (HYP) to 100 ± 0.0% in all other groups without significant differences among these groups. Similarly very high survival was reached in trial with HB with levels from 99.2 ± 1.3 (HYP, HYP1/1 and NOR) to 100 ± 0.0% (HYP ± ). HB fish showed best growth performance in NOR group reached final body weight (BW) 180.4 ± 2.3 g. Fish growth under different hyperoxic regimes was significantly reduced and final BW was 164.4 ± 7.6, 162.1 ± 12.2 and 151.7 ± 6.8 g in groups HY1/1, HYP ± and HYP, respectively. AC showed different preference for hyperoxic regimes as there were no significant difference in BW among NOR, HY1/1 and HYP± group with final values of 72.3 ± 11.3, 68.3 ± 8.4 and 77.1 ± 6.1g. Significantly reduced growth (BW 61.8 ± 6.8 g) was observed in HYP group. It is evident from present study that there are differences between pure bred Arctic char and hybrid in relation to hyperoxic regimes. The study was supported by projects 'CENAKVA' (No. CZ.1.05/2.1.00/01.0024), 'CENAKVA II' (No. LO1205 under the NPU I program), NAZV (QJ1510077) and GAJU (No. 060/2016/Z).

Keywords: recirculating aquaculture systems, Salmonidae, hyperoxia, abiotic factors

Procedia PDF Downloads 182

Authors: Helen Nelly Naiga, Jane Frances Zalwango, Saudah N. Kizito, Brian Agaba, Brenda N Simbwa, Maria Goretti Zalwango, Richard Migisha, Benon Kwesiga, Daniel Kadobera, Alex Ario Riolexus, Sarah Paige, Julie R. Harris

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Background: Traditional community beliefs and practices can facilitate the spread of Ebola virus during outbreaks. On September 20, 2022, Uganda declared a Sudan Virus Disease (SVD) outbreak after a case was confirmed in Mubende District. During September–November 2022, the outbreak spread to eight additional districts. We investigated the role of community beliefs and practices in the spread of SUDV in Uganda in 2022. Methods: A qualitative study was conducted in Mubende, Kassanda, and Kyegegwa districts in February 2023. We conducted nine focus group discussions (FGDs) and six key informant interviews (KIIs). FGDs included SVD survivors, household members of SVD patients, traditional healers, religious leaders, and community leaders. Key informants included community, political, and religious leaders, traditional healers, and health workers. We asked about community beliefs and practices to understand if and how they contributed to the spread of SUDV. Interviews were recorded, translated, transcribed, and analyzed thematically. Results: Frequently-reported themes included beliefs that the community deaths, later found to be due to SVD, were the result of witchcraft or poisoning. Key informants reported that SVD patients frequently first consulted traditional healers or spiritual leaders before seeking formal healthcare, and noted that traditional healers treated patients with signs and symptoms of SVD without protective measures. Additional themes included religious leaders conducting laying-on-of-hands prayers for SVD patients and symptomatic contacts, SVD patients and their symptomatic contacts hiding in friends’ homes, and exhumation of SVD patients originally buried in safe and dignified burials, to enable traditional burials. Conclusion: Multiple community beliefs and practices likely promoted SVD outbreak spread during the 2022 outbreak in Uganda. Engaging traditional and spiritual healers early during similar outbreaks through risk communication and community engagement efforts could facilitate outbreak control. Targeted community messaging, including clear biological explanations for clusters of deaths and information on the dangers of exhuming bodies of SVD patients, could similarly facilitate improved control in future outbreaks in Uganda.

Keywords: Ebola, Sudan virus, outbreak, beliefs, traditional

Procedia PDF Downloads 55

Authors: Ruwani N. Nugara, Masashi Inafuku, Kensaku Takara, Hironori Iwasaki, Hirosuke Oku

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Pteryxin from the partially purified hexane phase (HP) of Peucedanum japonicum Thunb (PJT) was identified as the active compound related to anti-obesity. Thus, in this study we investigated the mechanisms related to anti-obesity activity in-vitro. The HP was fractionated, and effect on the triglyceride (TG) content was evaluated in 3T3-L1 and HepG2 cells. Comprehensive spectroscopic analyses were used to identify the structure of the active compound. The dose dependent effect of active constituent on the TG content, and the gene expressions related to adipogenesis, fatty acid catabolism, energy expenditure, lipolysis and lipogenesis (20 μg/mL) were examined in-vitro. Furthermore, higher dosage of pteryxin (50μg/mL) was tested against 20μg/mL in 3T3-L1 adipocytes. The mRNA were subjected to SOLiD next generation sequencer and the obtained data were analyzed by Ingenuity Pathway Analysis (IPA). The active constituent was identified as pteryxin, a known compound in PJT. However, its biological activities against obesity have not been reported previously. Pteryxin dose dependently suppressed TG content in both 3T3-L1 adipocytes and HepG2 hepatocytes (P < 0.05). Sterol regulatory element-binding protein-1 (SREBP1 c), Fatty acid synthase (FASN), and acetyl-CoA carboxylase-1 (ACC1) were downregulated in pteryxin-treated adipocytes (by 18.0, 36.1 and 38.2%; P < 0.05, respectively) and hepatocytes (by 72.3, 62.9 and 38.8%, respectively; P < 0.05) indicating its suppressive effects on fatty acid synthesis. The hormone-sensitive lipase (HSL), a lipid catabolising gene was upregulated (by 15.1%; P < 0.05) in pteryxin-treated adipocytes suggesting improved lipolysis. Concordantly, the adipocyte size marker gene, paternally expressed gene1/mesoderm specific transcript (MEST) was downregulated (by 42.8%; P < 0.05), further accelerating the lipolytic activity. The upregulated trend of uncoupling protein 2 (UCP2; by 77.5%; P < 0.05) reflected the improved energy expenditure due to pteryxin. The 50μg/mL dosage of pteryxin completely suppressed PPARγ, MEST, SREBP 1C, HSL, Adiponectin, Fatty Acid Binding Protein (FABP) 4, and UCP’s in 3T3-L1 adipocytes. The IPA suggested that pteryxin at 20μg/mL and 50μg/mL suppress obesity in two different pathways, whereas the WNT signaling pathway play a key role in the higher dose of pteryxin in preadipocyte stage. Pteryxin in PJT play the key role in regulating lipid metabolism related gene network and improving energy production in vitro. Thus, the results suggests pteryxin as a new natural compound to be used as an anti-obesity drug in pharmaceutical industry.

Keywords: obesity, peucedanum japonicum thunb, pteryxin, food science

Procedia PDF Downloads 453

Authors: Swati Tomar, Sunil Kumar Gupta

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Anammox is a novel and promising technology that has changed the traditional concept of biological nitrogen removal. The process facilitates direct oxidation of ammonical nitrogen under anaerobic conditions with nitrite as an electron acceptor without the addition of external carbon sources. The present study investigated the feasibility of anammox hybrid reactor (AHR) combining the dual advantages of suspended and attached growth media for biodegradation of ammonical nitrogen in wastewater. The experimental unit consisted of 4 nos. of 5L capacity AHR inoculated with mixed seed culture containing anoxic and activated sludge (1:1). The process was established by feeding the reactors with synthetic wastewater containing NH4-H and NO2-N in the ratio 1:1 at HRT (hydraulic retention time) of 1 day. The reactors were gradually acclimated to higher ammonium concentration till it attained pseudo steady state removal at a total nitrogen concentration of 1200 mg/l. During this period, the performance of the AHR was monitored at twelve different HRTs varying from 0.25-3.0 d with increasing NLR from 0.4 to 4.8 kg N/m3d. AHR demonstrated significantly higher nitrogen removal (95.1%) at optimal HRT of 1 day. Filter media in AHR contributed an additional 27.2% ammonium removal in addition to 72% reduction in the sludge washout rate. This may be attributed to the functional mechanism of filter media which acts as a mechanical sieve and reduces the sludge washout rate many folds. This enhances the biomass retention capacity of the reactor by 25%, which is the key parameter for successful operation of high rate bioreactors. The effluent nitrate concentration, which is one of the bottlenecks of anammox process was also minimised significantly (42.3-52.3 mg/L). Process kinetics was evaluated using first order and Grau-second order models. The first-order substrate removal rate constant was found as 13.0 d-1. Model validation revealed that Grau second order model was more precise and predicted effluent nitrogen concentration with least error (1.84±10%). A new mathematical model based on mass balance was developed to predict N2 gas in AHR. The mass balance model derived from total nitrogen dictated significantly higher correlation (R2=0.986) and predicted N2 gas with least error of precision (0.12±8.49%). SEM study of biomass indicated the presence of the heterogeneous population of cocci and rod shaped bacteria of average diameter varying from 1.2-1.5 mm. Owing to enhanced NRE coupled with meagre production of effluent nitrate and its ability to retain high biomass, AHR proved to be the most competitive reactor configuration for dealing with nitrogen laden wastewater.

Keywords: anammox, filter media, kinetics, nitrogen removal

Procedia PDF Downloads 382

Authors: Mirjana Petronijević, Sanja Panić, Aleksandra Cvetanović, Branko Kordić, Nenad Grba

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Enzyme peroxidases are biological catalysts and play a major role in phenolic wastewater treatments and other environmental applications. The most studied species from the peroxidases family is horseradish peroxidase (HRP). In environmental processes, HRP could be used in its free or immobilized form. Enzyme immobilization onto solid support is performed to improve the enzyme properties, prolong its lifespan and operational stability and allow its reuse in industrial applications. One of the enzyme supports of a newer generation is magnetic particles (MPs). Fe₃O₄ MPs are the most widely pursued immobilization of enzymes owing to their remarkable advantages of biocompatibility and non-toxicity. Also, MPs can be easily separated and recovered from the water by applying an external magnetic field. On the other hand, metals and metal oxides are not suitable for the covalent binding of enzymes, so it is necessary to perform their surface modification. Fe₃O₄ MPs functionalization could be performed during the process of their synthesis if it takes place in the presence of plant extracts. Extracts of plant material, such as wild plants, herbs, even waste materials of the food and agricultural industry (bark, shell, leaves, peel), are rich in various bioactive components such as polyphenols, flavonoids, sugars, etc. When the synthesis of magnetite is performed in the presence of plant extracts, bioactive components are incorporated into the surface of the magnetite, thereby affecting its functionalization. In this paper, the suitability of bio-magnetite as solid support for covalent immobilization of HRP across glutaraldehyde was examined. The activity of immobilized HRP at different pH values (4-9) and temperatures (20-80°C) and reusability were examined. Bio-MP was synthesized by co-precipitation method from Fe(II) and Fe(III) sulfate salts in the presence of water extract of the Allium cepa peel. The water extract showed 81% of antiradical potential (according to DPPH assay), which is connected with the high content of polyphenols. According to the FTIR analysis, the bio-magnetite contains oxygen functional groups (-OH, -COOH, C=O) suitable for binding to glutaraldehyde, after which the enzyme is covalently immobilized. The immobilized enzyme showed high activity at ambient temperature and pH 7 (30 U/g) and retained ≥ 80% of its activity at a wide range of pH (5-8) and temperature (20-50°C). The HRP immobilized onto bio-MPs showed remarkable stability towards temperature and pH variations compared to the free enzyme form. On the other hand, immobilized HRP showed low reusability after the first washing cycle enzyme retains 50% of its activity, while after the third washing cycle retains only 22%.

Keywords: bio-magnetite, enzyme immobilization, water extracts, environmental protection

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Authors: Antonina A. Shumakova, Sergey A. Khotimchenko

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The relevance of research is associated, on the one hand, with an ever-increasing volume of production and the expansion of the scope of application of engineered nanomaterials (ENMs), and on the other hand, with the lack of sufficient scientific information on the nature of the interactions of nanoparticles (NPs) with components of biogenic and abiogenic origin. In particular, studying the effect of ENMs (TiO2 NPs, SiO2 NPs, Al2O3 NPs, fullerenol) on the toxicometric characteristics of common contaminants such as lead and cadmium is an important hygienic task, given the high probability of their joint presence in food products. Data were obtained characterizing a multidirectional change in the toxicity of model toxicants when they are co-administered with various types of ENMs. One explanation for this fact is the difference in the adsorption capacity of ENMs, which was further studied in in vitro studies. For this, a method was proposed based on in vitro modeling of conditions simulating the environment of the small intestine. It should be noted that the obtained data are in good agreement with the results of in vivo experiments: - with the combined administration of lead and TiO2 NPs, there were no significant changes in the accumulation of lead in rat liver; in other organs (kidneys, spleen, testes and brain), the lead content was lower than in animals of the control group; - studying the combined effect of lead and Al2O3 NPs, a multiple and significant increase in the accumulation of lead in rat liver was observed with an increase in the dose of Al2O3 NPs. For other organs, the introduction of various doses of Al2O3 NPs did not significantly affect the bioaccumulation of lead; - with the combined administration of lead and SiO2 NPs in different doses, there was no increase in lead accumulation in all studied organs. Based on the data obtained, it can be assumed that at least three scenarios of the combined effects of ENMs and chemical contaminants on the body: - ENMs quite firmly bind contaminants in the gastrointestinal tract and such a complex becomes inaccessible (or inaccessible) for absorption; in this case, it can be expected that the toxicity of both ENMs and contaminants will decrease; - the complex formed in the gastrointestinal tract has partial solubility and can penetrate biological membranes and / or physiological barriers of the body; in this case, ENMs can play the role of a kind of conductor for contaminants and, thus, their penetration into the internal environment of the body increases, thereby increasing the toxicity of contaminants; - ENMs and contaminants do not interact with each other in any way, therefore the toxicity of each of them is determined only by its quantity and does not depend on the quantity of another component. Authors hypothesized that the degree of adsorption of various elements on the surface of ENMs may be a unique characteristic of their action, allowing a more accurate understanding of the processes occurring in a living organism.

Keywords: absorption, cadmium, engineered nanomaterials, lead

Procedia PDF Downloads 87

Authors: Serap Gedikli, Hakan Çakmak, M. Buğra Güldiken, Duygu Yalnızoğlu

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Air, which is necessary for living things to survive; while it carries some useful substances in it, it can also carry foreign particles of different sizes that may be harmful to the health. All airborne organic substances of biological origin, including bacteria, fungi, fungal spores, viruses, pollen, and their components, are called "bioaerosols". Nowadays, everyone spends most of their time in closed areas such as home, workplace, school, etc. Although it is known that outdoor air pollution affects health, it is not known that indoor air pollution has harmful effects in terms of health. In this study, indoor air microbial load and SARS-CoV-2 virus cleaning efficiency of Froumann brand air cleaners were studied. This work in 300 m³, 600 m³, and 1000 m³ completely closed areas without any air circulation with Froumann N80, N90, and N100 air-cleaning devices. Analyzes were performed for both areas at 60 minutes before and after the device was operated using a particle measuring device (Particles Plus 7302) and an air sampler (Mas-100 ECO). The measurements were taken by placing the test equipment 1.5-2 m away from the air cleaner. At the same time, the efficiency of the HEPA filter was evaluated by taking samples from the air outlet point of the HEPA filter using the air sampling device (Mas-100 ECO) after the device was started. Nutrient agar and malt agar are used as total mesophilic bacteria and total fungi. The number of colony-forming units per m³ (cfu/m³) was calculated by counting colonies in Petri dishes after incubation for 48 hours at 37°C for bacteria and 72 hours at 30°C for fungi. The change in the number of colonies and the decrease in the microbial load was calculated as a percentage value. SARS-CoV-2 activity analysis studies were carried out by İnönü University Microbiology Department in accordance with the World Health Organization regulations. Finally, the HEPA filter in the devices used was taken and kept under a certain temperature and humidity, and the change in the microbial load on it was monitored over a 6-month period. At the end of the studies, a 91%-94% reduction was determined in the total mesophilic bacteria count of Frouman brand N80, N90, and N100 model air cleaners. A decrease of 94%-96% was detected in the total number of yeast/molds. HEPA filter efficiency was evaluated, and at the end of the analysis, 98% of the bacterial load and approximately 100% of yeast/mold load at the HEPA filter air outlet point were decreased. According to the SARS- CoV-2 analysis results, when the device is operating at the medium airflow level 3, it can filter virus-carrying aerosols by 99%. As a result, it was determined that the Froumann model air cleaner was effective in controlling and reducing the microbial load in the indoor air.

Keywords: HEPA filter, indoor air quality, microbial load, SARS-CoV-2

Procedia PDF Downloads 204

Authors: Bruno C. Schimming, Leandro L. Martins, PatríCia F. F. Pinheiro, Raquel F. Domeniconi, FabríCio S. Oliveira

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The agouti is a wildlife rodent that can be used as an alternative source of animal protein and this species has been raised in captivity in Brazil with the aim of providing meat. Thus, the knowledge of their reproductive biology and morphology of the reproductive organs is important. The objective of this study was to describe the morphology of epididymis in the Azara’s agouti, by light microscopy. Samples of epididymis were obtained from five adult Azara’s agouti (Dasyprocta azarae) during castration surgery performed at the Municipal Zoo of Catanduva, Brazil. Fragments of the epididymal regions (initial segment, caput, corpus and cauda) were collected. The biological samples were immediately fixed in paraformaldehyde for 24 hours, followed by histologic procedures comprising embedding in ParaplastTM (Sigma, St. Louis, MO, USA), sections of 5 µm, and staining with HE and Masson’s trichrome. The epididymis was a highly convoluted tubule that links the testis to the vas deferens. The epithelium lining was pseudostratified columnar surrounded by a periductal stroma. The epithelium contains several cell types: principal, basal, apical, clear, and hallo cells. Principal cells were the most abundant cell type. There were observed also migratory cells named halo cells. The caput epididymis was divided into two different regions: initial segment and caput. The initial segment has a very wide lumen, a high epithelium with conspicuous microvilli and the lumen was wide with exfoliated material. The other region of the caput epididymis, showed a lower epithelium when compared with the initial segment, large amounts of spermatozoa in the lumen, and a cytoplasmic vacuolization. This region presented many narrows cells. Many spermatozoa appeared in the lumen of corpus epididymis. The cauda region had a lower epithelium than the other epididymal regions in the agouti. The cauda epithelium presented plicae protruding into the lumen. Large amounts of spermatozoa are also present in the lumen. Small microvilli uniformly arranged so as to form a kind of “brush border” are observed on the apical surface of the cauda epithelium. The pattern of the epithelium lining the duct of the agouti epididymis does not differ greatly from that reported to other mammals, such as domestic and wildlife animals. These findings can cooperate with future investigations especially those related to rational exploration of these animals. All experimental procedures were approved by the institutional ethics committee (CEUA 796/2015). This study was supported by FAPESP (Grants 2015/23822-1).

Keywords: wildlife, testis excurrent ducts, epididymis, morphology

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Authors: Marc Dietrich

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Youth cultures like Punk as much as media relevant to the specific scenes associated with them offer ample opportunity for young people or juvenile adults to construct their personal identities. However, developmental psychology has largely neglected such identity construction processes during the last decades. Such was not always the case: Early developmental psychologists intensely studied youth cultures and their meaningful objects and media in the early 20th century but lost interest when cultural studies and the social sciences occupied the field after World War II. Our project Constructions of Juvenility and Generation(ality), funded by the German Federal Ministry for Education and Research, reintegrates the study of youth cultures and their meaningful objects and media in a developmental psychology perspective. We present an empirical study of the ways in which youth, juvenility, and generation (ality) are constructed and negotiated in underground media like punk fanzines (a portmanteau of fan and magazine), including both semantic and aesthetic aspects of these construction processes within punk culture. The fanzine sample was accessed by the theoretical sampling strategy typical for GTM studies. Acknowledging fanzines as artful self-produced media by scene members for scene members, we conceptualize them as authentic documents of scene norms and values. Drawing on an analysis of both text and (cover) images in Punk fanzines published in Germany (and within a sample dating from 1981 until 2015) using a novel Visual Grounded Theory approach, we found that: a) Juvenility is a highly contested concept in punk culture. Its semantic quality and valuation varies with the perspectives present within the culture (e.g. embryo punks versus older punks); b) Juvenility is constructed as having energy and being socio-critical that does not depend on biological age; c) Juvenility is regarded not an ideal per se in German Punk culture; Punk culture constructs old age in a largely positive way (e.g., as marker of being real and a historical innovator); d) Juvenility is constructed as a habit that should be kept for life as it is constantly adapted to individual biographical trajectories like specific job situations or having a family. Consequently, identity negotiation as documented in the zines attempts to balance subculturally driven perspectives on life and society with the pragmatic requirements of a bourgeois life. The proposed paper will present the main results of this large-scale study of punk fanzines and show how developmental psychology perspectives as represented in the novel methodology applied in it can advance the study of youth cultures.

Keywords: construction of juvenility, developmental psychology, visual GTM, youth culture, fanzines

Procedia PDF Downloads 292

Authors: Benoit Atge, Alice Dhersin, Oscar Da Silva Cacao, Beatrice Martinez, Dominique Ducint, Catherine Verdun-Esquer, Isabelle Baldi, Mathieu Molimard, Antoine Villa, Mireille Canal-Raffin

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Introduction: Healthcare workers' exposure to antineoplastic drugs (AD) is a burning issue for occupational medicine practitioners. Biological monitoring of occupational exposure (BMOE) is an essential tool for assessing AD contamination of healthcare workers. In addition to BMOE, surface sampling is a useful tool in order to understand how workers get contaminated, to identify sources of environmental contamination, to verify the effectiveness of surface decontamination way and to ensure monitoring of these surfaces. The objective of this work was to develop a complete tool including a kit for surface sampling and a quantification analytical method for AD traces detection. The development was realized with the three following criteria: the kit capacity to sample in every professional environment (healthcare services, veterinaries, etc.), the detection of very low AD traces with a validated analytical method and the easiness of the sampling kit use regardless of the person in charge of sampling. Material and method: AD mostly used in term of quantity and frequency have been identified by an analysis of the literature and consumptions of different hospitals, veterinary services, and home care settings. The kind of adsorbent device, surface moistening solution and mix of solvents for the extraction of AD from the adsorbent device have been tested for a maximal yield. The AD quantification was achieved by an ultra high-performance liquid chromatography method coupled with tandem mass spectrometry (UHPLC-MS/MS). Results: With their high frequencies of use and their good reflect of the diverse activities through healthcare, 15 AD (cyclophosphamide, ifosfamide, doxorubicin, daunorubicin, epirubicin, 5-FU, dacarbazin, etoposide, pemetrexed, vincristine, cytarabine, methothrexate, paclitaxel, gemcitabine, mitomycin C) were selected. The analytical method was optimized and adapted to obtain high sensitivity with very low limits of quantification (25 to 5000ng/mL), equivalent or lowest that those previously published (for 13/15 AD). The sampling kit is easy to use, provided with a didactic support (online video and protocol paper). It showed its effectiveness without inter-individual variation (n=5/person; n= 5 persons; p=0,85; ANOVA) regardless of the person in charge of sampling. Conclusion: This validated tool (sampling kit + analytical method) is very sensitive, easy to use and very didactic in order to control the chemical risk brought by AD. Moreover, BMOE permits a focal prevention. Used in routine, this tool is available for every intervention of occupational health.

Keywords: surface contamination, sampling kit, analytical method, sensitivity

Procedia PDF Downloads 132

Authors: John T. Leman, James Gibson, Peter J. Bonitatibus

Abstract:

There have been no potential clinically translatable developments of intravenous CT contrast materials over decades, and iodinated contrast agents (ICA) remain the only FDA-approved media for CT. Small molecule ICA used to highlight vascular anatomy have weak CT signals in large-to-obese patients due to their rapid redistribution from plasma into interstitial fluid, thereby diluting their intravascular concentration, and because of a mismatch of iodine’s K-edge and the high kVp settings needed to image this patient population. The use of ICA is also contraindicated in a growing population of renally impaired patients who are hypersensitive to these contrast agents; a transformative intravenous contrast agent with improved capabilities is urgently needed. Tantalum oxide nanoparticles (TaO NPs) with zwitterionic siloxane polymer coatings have high potential as clinically translatable general-purpose CT contrast agents because of (1) substantially improved imaging efficacy compared to ICA in swine/phantoms emulating medium-sized and larger adult abdomens and superior thoracic vascular contrast enhancement of thoracic arteries and veins in rabbit, (2) promising biological safety profiles showing near-complete renal clearance and low tissue retention at 3x anticipated clinical dose (ACD), and (3) clinically acceptable physiochemical parameters as concentrated bulk solutions(250-300 mgTa/mL). Here, we review requirements for general-purpose intravenous CT contrast agents in terms of patient safety, X-ray attenuating properties and contrast-producing capabilities, and physicochemical and pharmacokinetic properties. We report the current status of a TaO NP-based contrast agent, including chemical process technology developments and results of newly defined scaled-up processes for NP synthesis and purification, yielding reproducible formulations with appropriate size and concentration specifications. We discuss recent results of recent pre-clinical in vitro immunology, non-GLP high dose tolerability in rats (10x ACD), non-GLP long-term biodistribution in rats at 3x ACD, and non-GLP repeat dose in rats at ACD. We also include a discussion of NP characterization, in particular size-stability testing results under accelerated conditions (37C), and insights into TaO NP purity, surface structure, and bonding of the zwitterionic siloxane polymer coating by multinuclear (1H, 13C, 29Si) and multidimensional (2D) solution NMR spectroscopy.

Keywords: nanoparticle, imaging, diagnostic, process technology, nanoparticle characterization

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Authors: Dalita G. S. M. Cavalcante, Elton A. P. dos Reis, Andressa S. Gomes, Caroline S. Danna, Leandra Ernest Kerche-Silva, Eidi Yoshihara, Aldo E. Job

Abstract:

In order to minimize environmental impacts, a composite was developed from mixture of leather shavings (LE) with natural rubber (NR), which patent is already deposited. The new material created can be used in applications such as floors e heels for shoes. Besides these applications, the aim is to use this new material for the production of products for the textile industry, such as boots, gloves and bags. But the question arises, as to biocompatibility of this new material. This is justified because the structure of the leather shavings has chrome. The trivalent chromium is usually not toxic, but the hexavalent chromium can be highly toxic and genotoxic for living beings, causing damage to the DNA molecule and contributing to the formation of cancer. Based on this, the objective of this study is evaluate the possible genotoxic effects of the new composite, using as system - test two cell lines (MRC-5 and CHO-K1) by comet assay. For this, the production of the composite was performed in three proportions: for every 100 grams of NR was added 40 (E40), 50 (E50) or 60 (E60) grams of LE. The latex was collected from the rubber tree (Hevea brasiliensis). For vulcanization of the NR, activators and accelerators were used. The two cell lines were exposed to the new composite in its three proportions using elution method, that is, cells exposed to liquid extracts obtained from the composite for 24 hours. For obtaining the liquid extract, each sample of the composite was crushed into pieces and mixed with an extraction solution. The quantification of total chromium and hexavalent chromium in the extracts were performed by Optical Emission Spectrometry by Inductively Coupled Plasma (ICP-OES). The levels of DNA damage in cells exposed to both extracts were monitored by alkaline version of the comet assay. The results of the quantification of metals in ICP-OES indicated the presence of total chromium in different extracts, but were not detected presence of hexavalent chromium in any extract. Through the comet assay were not found DNA damage of the CHO-K1 cells exposed to both extracts. As for MRC-5, was found a significant increase in DNA damage in cells exposed to E50 and E60. Based on the above data, it can be asserted that the extracts obtained from the composite were highly genotoxic for MRC-5 cells. These biological responses do not appear to be related to chromium metal, since there was a predominance of trivalent chromium in the extracts, indicating that during the production process of the new composite, there was no formation of hexavalent chromium. In conclusion it can infer that the leather shavings containing chromium can be reused, thereby reducing the environmental impacts of this waste. Already on the composite indicates to its incorporation in applications that do not aim at direct contact with the human skin, and it is suggested the chain of composite production be studied, in an attempt to make it biocompatible so that it may be safely used by the textile industry.

Keywords: cell line, chrome, genotoxicity, leather, natural rubber

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Authors: Lina Wu

Abstract:

The excessive discharge of nitrogen in sewage greatly intensifies the eutrophication of water bodies and poses a threat to water quality. Nitrogen pollution control has become a global concern. Currently, the problem of water pollution in China is still not optimistic. As a typical high ammonia nitrogen organic wastewater, landfill leachate is more difficult to treat than domestic sewage because of its complex water quality, high toxicity, and high concentration.Many studies have shown that the autotrophic anammox bacteria in nature can combine nitrous and ammonia nitrogen without carbon source through functional genes to achieve total nitrogen removal, which is very suitable for the removal of nitrogen from leachate. In addition, the process also saves a lot of aeration energy consumption than the traditional nitrogen removal process. Therefore, anammox plays an important role in nitrogen conversion and energy saving. The process composed of short-range nitrification and denitrification coupled an ammo ensures the removal of total nitrogen and improves the removal efficiency, meeting the needs of the society for an ecologically friendly and cost-effective nutrient removal treatment technology. Continuous flow process for treating late leachate [an up-flow anaerobic sludge blanket reactor (UASB), anoxic/oxic (A/O)–anaerobic ammonia oxidation reactor (ANAOR or anammox reactor)] has been developed to achieve autotrophic deep nitrogen removal. In this process, the optimal process parameters such as hydraulic retention time and nitrification flow rate have been obtained, and have been applied to the rapid start-up and stable operation of the process system and high removal efficiency. Besides, finding the characteristics of microbial community during the start-up of anammox process system and analyzing its microbial ecological mechanism provide a basis for the enrichment of anammox microbial community under high environmental stress. One research developed partial nitrification-Anammox (PN/A) using an internal circulation (IC) system and a biological aerated filter (BAF) biofilm reactor (IBBR), where the amount of water treated is closer to that of landfill leachate. However, new high-throughput sequencing technology is still required to be utilized to analyze the changes of microbial diversity of this system, related functional genera and functional genes under optimal conditions, providing theoretical and further practical basis for the engineering application of novel anammox system in biogas slurry treatment and resource utilization.

Keywords: nutrient removal and recovery, leachate, anammox, partial nitrification

Procedia PDF Downloads 51

Authors: Carmela Nardelli, Laura Iaffaldano, Valentina Capobianco, Antonietta Tafuto, Maddalena Ferrigno, Angela Capone, Giuseppe Maria Maruotti, Maddalena Raia, Rosa Di Noto, Luigi Del Vecchio, Pasquale Martinelli, Lucio Pastore, Lucia Sacchetti

Abstract:

Maternal obesity and nutrient excess in utero increase the risk of future metabolic diseases in the adult life. The mechanisms underlying this process are probably based on genetic, epigenetic alterations and changes in foetal nutrient supply. In mammals, the placenta is the main interface between foetus and mother, it regulates intrauterine development, modulates adaptive responses to sub optimal in uterus conditions and it is also an important source of human amniotic mesenchymal stem cells (hA-MSCs). We previously highlighted a specific microRNA (miRNA) profiling in amnion from obese (Ob) pregnant women, here we compared the miRNA expression profile of hA-MSCs isolated from (Ob) and control (Co) women, aimed to search for any alterations in metabolic pathways that could predispose the new-born to the obese phenotype. Methods: We isolated, at delivery, hA-MSCs from amnion of 16 Ob- and 7 Co-women with pre-pregnancy body mass index (mean/SEM) 40.3/1.8 and 22.4/1.0 kg/m2, respectively. hA-MSCs were phenotyped by flow cytometry. Globally, 384 miRNAs were evaluated by the TaqMan Array Human MicroRNA Panel v 1.0 (Applied Biosystems). By the TargetScan program we selected the target genes of the miRNAs differently expressed in Ob- vs Co-hA-MSCs; further, by KEGG database, we selected the statistical significant biological pathways. Results: The immunophenotype characterization confirmed the mesenchymal origin of the isolated hA-MSCs. A large percentage of the tested miRNAs, about 61.4% (232/378), was expressed in hA-MSCs, whereas 38.6% (146/378) was not. Most of the expressed miRNAs (89.2%, 207/232) did not differ between Ob- and Co-hA-MSCs and were not further investigated. Conversely, 4.8% of miRNAs (11/232) was higher and 6.0% (14/232) was lower in Ob- vs Co-hA-MSCs. Interestingly, 7/232 miRNAs were obesity-specific, being expressed only in hA-MSCs isolated from obese women. Bioinformatics showed that these miRNAs significantly regulated (P<0.001) genes belonging to several metabolic pathways, i.e. MAPK signalling, actin cytoskeleton, focal adhesion, axon guidance, insulin signaling, etc. Conclusions: Our preliminary data highlight an altered miRNA profile in Ob- vs Co-hA-MSCs and suggest that an epigenetic miRNA-based mechanism of gene regulation could affect pathways involved in placental growth and function, thereby potentially increasing the newborn’s risk of metabolic diseases in the adult life.

Keywords: hA-MSCs, obesity, miRNA, biosystem

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Authors: Ajuna B. Henry

Abstract:

In recent decades, climate change has demanded biological pesticides; more Bt strains are being discovered worldwide, some containing novel insecticidal genes while others have been modified through molecular approaches for increased yield, toxicity, and wider host target. In this study, B. thuringiensis strain AH-2 (Bt-2) was isolated from the soil and tested for insecticidal activity against Aphis gossypii (Hemiptera: Aphididae) and Lepidoptera insect pests: fall webworm (Hyphantria cunea) and diamondback moth (Plutella xylostella). A commercial strain B. thuringiensis subsp. kurstaki (Btk), and a chemical pesticide, imidacloprid (for Hemiptera) and chlorantraniliprole (for Lepidoptera), were used as positive control and the same media (without bacterial inoculum) as a negative control. For aphidicidal activity, Bt-2 caused a mortality rate of 70.2%, 78.1% or 88.4% in third instar nymphs of A. gossypii (3N) at 10%, 25% or 50% culture concentrations, respectively. Moreover, Bt-2 was effectively produced in cost-effective (PB) supplemented with either glucose (PBG) or sucrose (PBS) and maintained high aphicidal efficacy with 3N mortality rates of 85.9%, 82.9% or 82.2% in TSB, PBG or PBS media, respectively at 50% culture concentration. Bt-2 also suppressed adult fecundity by 98.3% compared to only 65.8% suppression by Btk at similar concentrations but was slightly lower than chemical treatment, which caused 100% suppression. Partial purification of 60 – 80% (NH4)2SO4 fraction of Bt-2 aphicidal proteins purified on anion exchange (DEAE-FF) column revealed a 105 kDa aphicidal protein with LC50 = 55.0 ng/µℓ. For Lepidoptera pests, chemical pesticide, Bt-2, and Btk cultures, mortality of 86.7%, 60%, and 60% in 3rd instar larvae of P. xylostella, and 96.7%, 80.0%, and 93.3% in 6th instar larvae of H. cunea, after 72h of exposure. When the entomopathogenic strains were cultured in a cost-effective PBG or PBS, the insecticidal activity in all strains was not significantly different compared to the use of a commercial medium (TSB). Bt-2 caused a mortality rate of 60.0%, 63.3%, and 50.0% against P. xylostella larvae and 76.7%, 83.3%, and 73.3% against H. cunea when grown in TSB, PBG, and PBS media, respectively. Bt-2 (grown in cost-effective PBG medium) caused a dose-dependent toxicity of 26.7%, 40.0%, and 63.3% against P. xylostella and 46.7%, 53.3%, and 76.7% against H. cunea at 10%, 25% and 50% culture concentration, respectively. The partially purified Bt-2 insecticidal proteins fractions F1, F2, F3, and F4 (extracted at different ratios of organic solvent) caused low toxicity (50.0%, 40.0%, 36.7%, and 30.0%) against P. xylostella and relatively high toxicity (56.7%, 76.7%, 66.7%, and 63.3%) against H. cunea at 100 µg/g of artificial diets. SDS-PAGE analysis revealed that a128kDa protein is associated with toxicity of Bt-2. Our result demonstrates a medium and strong larvicidal activity of Bt-2 against P. xylostella and H. cunea, respectively. Moreover, Bt-2 could be potentially produced using a cost-effective PBG medium which makes it an effective alternative biocontrol strategy to reduce chemical pesticide application.

Keywords: biocontrol, insect pests, larvae/nymph mortality, cost-effective media, aphis gossypii, plutella xylostella, hyphantria cunea, bacillus thuringiensi

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