Search results for: enzymatic fermentation

Authors: Ozcan B., Koca B., Tuzcuoglu E., Cavusoglu S., Efe A., Bayraktar S.

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A washing machine is mainly used for removing any types of dirt and stains and also eliminating malodorous substances from textile surfaces. Stains originate from various sources from the human body to environmental contamination. Therefore, there are various methods for removing them. They are roughly classified into four different groups: oily (greasy) stains, particulate stains, enzymatic stains and bleachable (oxidizable) stains. Oily stains on clothes surfaces are a common result of being in contact with organic substances of the human body (e.g. perspiration, skin shedding and sebum) or by being exposed to an oily environmental pollutant (e.g. oily foods). Studies showed that human sebum is major component of oily soil found on the garments, and if it is aged under the several environmental conditions, it can generate obstacle yellow stains on the textile surface. In this study, a parametric study was carried out to investigate the key factors affecting the cleaning performance (specifically sebum removal performance) of a washing machine. These parameters are mechanical agitation percentage of tumble, consumed water and total washing period. A full factorial design of the experiment is used to capture all the possible parametric interactions using Minitab 2021 statistical program. Tests are carried out with commercial liquid detergent and 2 different types of sebum-soiled cotton and cotton + polyester fabrics. Parametric results revealed that for both test samples, increasing the washing time and the mechanical agitation could lead to a much better removal result of sebum. However, for each sample, the water amount had different outcomes. Increasing the water amount decreases the performance of cotton + polyester fabrics, while it is favorable for cotton fabric. Besides this, it was also discovered that the type of textile can greatly affect the sebum removal performance. Results showed that cotton + polyester fabrics are much easier to clean compared to cotton fabric

Keywords: laundry, washing machine, low-temperature washing, cold wash, washing efficiency index, sustainability, cleaning performance, stain removal, oily soil, sebum, yellowing

Procedia PDF Downloads 134

Authors: Narayan Moger, Divya B., Preethi Jambagi, Krishnaveni C. K., Apsana M. R., B. R. Patil, Basvaraj Bagewadi

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Salinity is one of the major threats to world food security. Considering the requirement for salt tolerant crop plants in the present study was undertaken to clone and transferred the salt tolerant ectA gene from marine ecosystem into agriculture crop system to impart salinity tolerance. Ectoine is the compatible solute which accumulates in the cell membrane, is known to be involved in salt tolerance activity in most of the Halophiles. The present situation is insisting to development of salt tolerant transgenic lines to combat abiotic stress. In this background, the investigation was conducted to develop transgenic tomato lines by cloning and transferring of ectA gene is an ectoine derivative capable of enzymatic action for the production of acetyl-diaminobutyric acid. The gene ectA is involved in maintaining the osmotic balance of plants. The PCR amplified ectA gene (579bp) was cloned into T/A cloning vector (pTZ57R/T). The construct pDBJ26 containing ectA gene was sequenced by using gene specific forward and reverse primers. Sequence was analyzed using BLAST algorithm to check similarity of ectA gene with other isolates. Highest homology of 99.66 per cent was found with ectA gene sequences of isolates Halomonas elongata with the available sequence information in NCBI database. The ectA gene was further sub cloned into pRI101-AN plant expression vector and transferred into E. coli DH5α for its maintenance. Further pDNM27 was mobilized into A. tumefaciens LBA4404 through tri-parental mating system. The recombinant Agrobacterium containing pDNM27 was transferred into tomato plants through In planta plant transformation method. Out of 300 seedlings, co-cultivated only twenty-seven plants were able to well establish under the greenhouse condition. Among twenty-seven transformants only twelve plants showed amplification with gene specific primers. Further work must be extended to evaluate the transformants at T1 and T2 generations for ectoine accumulation, salinity tolerance, plant growth and development and yield.

Keywords: salinity, computable solutes, ectA, transgenic, in planta transformation

Procedia PDF Downloads 76

Authors: Marwa Brahim, Nicolas Brosse, Nadia Boussetta, Nabil Grimi, Eugene Vorobiev

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Rapeseed plant is an established product in France which is mainly dedicated to oil production. However, the economic potential of residues from this industry (rapeseed hulls, rapeseed cake, rapeseed straw etc.), has not been fully exploited. Currently, only low-grade applications are found in the market. As a consequence, it was deemed of interest to develop a technological platform aiming to convert rapeseed residues into value- added products. Specifically, a focus is given on the conversion of rapeseed straw into valuable molecules (e.g. lignin, glucose). Existing pretreatment technologies have many drawbacks mainly the production of sugar degradation products that limit the effectiveness of saccharification and fermentation steps in the overall scheme of the lignocellulosic biorefinery. In addition, the viability of fractionation strategies is a challenge in an environmental context increasingly standardized. Hence, the need to find cleaner alternatives with comparable efficiency by implementing physical phenomena that could destabilize the structural integrity of biomass without necessarily using chemical solvents. To meet environmental standards increasingly stringent, the present work aims to study the new pretreatment strategies involving lower consumption of chemicals with an attenuation of the severity of the treatment. These strategies consist on coupling physical treatments either high voltage electrical discharges or ultrasounds to conventional chemical pretreatments (soda and organosolv). Ultrasounds treatment is based on the cavitation phenomenon, and high voltage electrical discharges cause an electrical breakdown accompanied by many secondary phenomena. The choice of process was based on a technological feasibility study taking into account the economic profitability of the whole chain after products valorization. Priority was given to sugars valorization into bioethanol and lignin sale.

Keywords: high voltage electrical discharges, organosolv, pretreatment strategies, rapeseed straw, soda, ultrasounds

Procedia PDF Downloads 359

Authors: Mohamed Benaddou, Mohammed Diouri

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country, such as Morocco, generates a high quantity of agricultural and food industry residues. A large portion of these residues is disposed of by burning or landfilling. The valorization of this waste biomass as feed is an interesting alternative because it is therefore considered among the best sources of cheap carbohydrates. However, its nutritional yield without any pre-treatment is very low because lignin protects cellulose, the carbohydrate used as a source of energy by ruminants. Fungal treatment is an environmentally friendly, easy and inexpensive method. This study investigated the treatment of wheat straw (WS), cedar sawdust (CS) and olive pomace (OP) with fungi selected according to the source of Carbon for improving its digestibility. Two were selected in a culture medium in which cellulose was the only source of Carbon: Cosmospora Viridescens (C.vir) and Penicillium crustosum (P.crus), two were selected in a culture medium in which lignin is the only source of Carbon: Fusarium oxysporum (F.oxy) and Fusarium sp. (F. Sp), and two in a culture medium where cellulose and lignin are the two sources of Carbon at the same time: Fusarium solani (F. solani) and Penicillium chrysogenum (P.chryso). P.chryso degraded more CS cellulose. It is very important to notice that the delignification by F. Solani reached 70% after 12 weeks of treatment of wheat straw. Ligninase enzymatic was detected in F.solani, F.sp, F.oxysporum, which made it possible to delignify the treated substrates. Delignification by C.vir is negligible in all three substrates after 12 weeks of treatment. P.crus and P.chryso degraded the lignin very slightly in WC (it did not exceed 12% after 12 weeks of treatment) but in OP this delignification is slight reaching 25% and 13% for P.chryso and P.crus successively. P.chryso allowed 30% degradation of lignin from 4 weeks of treatment. The degradation of the lignin was able to reach the maximum within 8 weeks of treatment for most of the fungi except F. solani who continued the treatment after this period. Digestibility variation (IVTD.variation) is highly very significant from fungus to fungi, duration to time, substrate to substrate and its interactions (P <0.001). indeed, all the fungi increased digestibility after 12 weeks of treatment with a difference in the degree of this increase. F.solani and F.oxy increased digestibility more than the others. this digestibility exceeded 50% in CS and O.P but did not exceed 20% for WS after treatment with F.oxy. IVTD.Var was not exceeded 20% in W.S.cedar treated with P.chryso but reached 45% after 8 weeks of treatment in W.straw.

Keywords: lignin, cellulose, digestibility, fungi, treatment, lignocellulosic biomass

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Authors: Shilpi Malik, Ramneek Kaur, Archita Gupta, Deepshikha Yadav, Ashwani Mathur, Manisha Singh

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Glucosamine (GS) is the most abundant naturally occurring amino monosaccharide and is normally produced in human body via cellular glucose metabolism. It is regarded as the building block of cartilage matrix and is also an essential component of cartilage matrix repair mechanism. Besides that, it can also be explored for its prebiotic potential as many bacterial species are known to utilize the amino sugar by acquiring them to form peptidoglycans and lipopolysaccharides in the bacterial cell wall. Glucosamine can therefore be considered for its fermentation by bacterial species present in the gut. Current study is focused on exploring the potential of glucosamine as prebiotic. The studies were done to optimize considerable concentration of GS to reach GI tract and being fermented by the complex gut microbiota and food grade GS was added to various Simulated Fluids of Gastro-Intestinal Tract (GIT) such as Simulated Saliva, Gastric Fluid (Fast and Fed State), Colonic fluid, etc. to detect its degradation. Since it was showing increase in microbial growth (CFU) with time, GS was Further, encapsulated to increase its residential time in the gut, which exhibited improved resistance to the simulated Gut conditions. Moreover, prepared microspehres were optimized and characterized for their encapsulation efficiency and toxicity. To further substantiate the prebiotic activity of Glucosamine, studies were also performed to determine the effect of Glucosamine on the known probiotic bacterial species, i.e. Lactobacillus delbrueckii (MTCC 911) and Bifidobacteriumbifidum (MTCC 5398). Culture conditions for glucosamine will be added in MRS media in anaerobic tube at 0.20%, 0.40%, 0.60%, 0.80%, and 1.0%, respectively. MRS media without GS was included in this experiment as the control. All samples were autoclaved at 118° C for 15 min. Active culture was added at 5% (v/v) to each anaerobic tube after cooling to room temperature and incubated at 37° C then determined biomass and pH and viable count at incubation 18h. The experiment was completed in triplicate and the results were presented as Mean ± SE (Standard error).The experimental results are conclusive and suggest Glucosamine to hold prebiotic properties.

Keywords: gastro intestinal tract, microspheres, peptidoglycans, simulated fluid

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Authors: K. Sakthivel

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With the continuous increase in population worldwide, stress increased among agricultural peoples, so it is necessary to increase the yield of agro-products. But it is not possible to meet fully with the traditionally adopted ways of using pesticides and herbicides. Today, agriculture and horticulture has realized the need of tomorrow and opting for various technologies to get higher overall yield, quality agro-products. Most of today’s synthetic polymers are produced from petrochemical bi-products and are not biodegradable. Persistent polymers generate significant sources of environmental pollution, harming wildlife when they are disposed in nature. The disposal of non degradable plastic bags adversely affects human and wild life. Moreover incineration of plastic waste presents environmental issues as well, since it yields toxic emissions. Material incineration is also limited due to the difficulties to find accurate and economically viable outlets. In addition plastic recycling shows a negative eco balance due to the necessity in nearly all cases to wash the plastic waste as well as the energy consumption during the recycling process phases. As plastics represent a large part of the waste collection at the local regional and national levels institutions are aware of the significant savings that compostable or biodegradable materials would generate. Polylactic acid (PLA), which is one of the most important biocompatible polyesters that are derived from annually renewable biomass such as corn and wheat, has attracted much attention for automotive parts and also can be applied in agro textiles. The manufacturing method of PLA is the ring-opening polymerization of the dimeric cyclic ester of lactic acid, lactide. For the stereo complex PLA, we developed by the four unit processes, fermentation, separation, lactide conversion, and polymerization. Then the polymer is converted into mulching film and applied in agriculture field. PLA agro textiles have better tensile strength, tearing strength and with stand from UV rays than polyester agro textile and polypropylene-based products.

Keywords: biodegradation, environment, mulching film, PLA, technical textiles

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Authors: Noura El-Ahmady El-Naggar

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Among the antitumour drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product(1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen nutritional variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4.7H2O, NaCl and FeSO4. 7H2O) were screened using Plackett–Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age and agitation speed) were further optimized by the central composite face-centered design -response surface methodology. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase in the culture filtrate of Streptomyces olivaceus NEAE-119: Dextrose 3g, starch 20g, L-asparagine 10g, KNO3 1g, K2HPO4 1g, MgSO4.7H2O 0.1g, NaCl 0.1g, pH 7, temperature 37°C, agitation speed 200 rpm/min, inoculum size 4%, v/v, inoculum age 72 h and fermentation period 5 days.

Keywords: Streptomyces olivaceus NEAE-119, glutaminase free L-asparaginase, production, Plackett-Burman design, central composite face-centered design, 16S rRNA, scanning electron microscope

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Authors: Sofiqul Islam, V. Murugan, Prema Kumari, Hari

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Gentamicin is a broad spectrum water-soluble aminoglycoside antibiotics produced by the fermentation process of microorganism known as Micromonospora purpurea. It is widely used for the treatment of infection caused by both gram positive and gram negative bacteria. Gentamicin consists of a mixture of aminoglycoside components like C1, C1a, C2a, and C2. The molecular structure of Gentamicin and its related substances showed that it has lack of presence of chromophore group in the molecule due to which the detection of such components were quite critical and challenging. In this study, a simple Reversed Phase-High Performance Liquid Chromatographic (RP-HPLC) method using ultraviolet (UV) detector was developed and validated for quantification of the related substances present in Gentamicin drug substances. The method was achieved by using Thermo Scientific Hypersil Gold analytical column (150 x 4.6 mm, 5 µm particle size) with isocratic elution composed of methanol: water: glacial acetic acid: sodium hexane sulfonate in the ratio 70:25:5:3 % v/v/v/w as a mobile phase at a flow rate of 0.5 mL/min, column temperature was maintained at 30 °C and detection wavelength of 330 nm. The four components of Gentamicin namely Gentamicin C1, C1a, C2a, and C2 were well separated along with the related substance present in Gentamicin. The Limit of Quantification (LOQ) values were found to be at 0.0075 mg/mL. The accuracy of the method was quite satisfactory in which the % recovery was resulted between 95-105% for the related substances. The correlation coefficient (≥ 0.995) shows the linearity response against concentration over the range of Limit of Quantification (LOQ). Precision studies showed the % Relative Standard Deviation (RSD) values less than 5% for its related substance. The method was validated in accordance with the International Conference of Harmonization (ICH) guideline with various parameters like system suitability, specificity, precision, linearity, accuracy, limit of quantification, and robustness. This proposed method was easy and suitable for use for the quantification of related substances in routine analysis of Gentamicin formulations.

Keywords: reversed phase-high performance liquid chromatographic (RP-HPLC), high performance liquid chromatography, gentamicin, isocratic, ultraviolet

Procedia PDF Downloads 157

Authors: Wighens Ngoie Ilunga, Pamela J. Welz, Olewaseun O. Oyekola, Daniel Ikhu-Omoregbe

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Currently, most sludge from the wastewater treatment plants of edible oil factories is disposed to landfills, but landfill sites are finite and potential sources of environmental pollution. Production of biodiesel from wastewater sludge can contribute to energy production and waste minimization. However, conventional biodiesel production is energy and waste intensive. Generally, biodiesel is produced from the transesterification reaction of oils with alcohol (i.e., Methanol, ethanol) in the presence of a catalyst. Homogeneously catalysed transesterification is the conventional approach for large-scale production of biodiesel as reaction times are relatively short. Nevertheless, homogenous catalysis presents several challenges such as high probability of soap. The current study aimed to reuse wastewater sludge from the edible oil industry as a novel feedstock for both monounsaturated fats and bioethanol for the production of biodiesel. Preliminary results have shown that the fatty acid profile of the oilseed wastewater sludge is favourable for biodiesel production with 48% (w/w) monounsaturated fats and that the residue left after the extraction of fats from the sludge contains sufficient fermentable sugars after steam explosion followed by an enzymatic hydrolysis for the successful production of bioethanol [29% (w/w)] using a commercial strain of Saccharomyces cerevisiae. A novel nano-magnetic catalyst was synthesised from mineral processing alkaline tailings, mainly containing dolomite originating from cupriferous ores using a modified sol-gel. The catalyst elemental chemical compositions and structural properties were characterised by X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infra-red (FTIR) and the BET for the surface area with 14.3 m²/g and 34.1 nm average pore diameter. The mass magnetization of the nano-magnetic catalyst was 170 emu/g. Both the catalytic properties and reusability of the catalyst were investigated. A maximum biodiesel yield of 78% was obtained, which dropped to 52% after the fourth transesterification reaction cycle. The proposed approach has the potential to reduce material costs, energy consumption and water usage associated with conventional biodiesel production technologies. It may also mitigate the impact of conventional biodiesel production on food and land security, while simultaneously reducing waste.

Keywords: biodiesel, bioethanol, edible oil wastewater sludge, nano-magnetism

Procedia PDF Downloads 138

Authors: Goksen Arik, Mihriban Korukluoglu

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Lactic acid bacteria (LAB) and some yeast species are common microorganisms found in dairy products and most of them are responsible for the fermentation of foods. Such cultures are isolated and used as a starter culture in the food industry because of providing standardisation of the final product during the food processing. Choice of starter culture is the most important step for the production of fermented food. Isolated LAB and yeast cultures which have the ability to create a biofilm layer can be preferred as a starter in the food industry. The biofilm formation could be beneficial to extend the period of usage time of microorganisms as a starter. On the other hand, it is an undesirable property in pathogens, since biofilm structure allows a microorganism become more resistant to stress conditions such as antibiotic presence. It is thought that the resistance mechanism could be turned into an advantage by promoting the effective microorganisms which are used in the food industry as starter culture and also which have potential to stimulate the gastrointestinal system. Development of the biofilm layer is observed in some LAB and yeast strains. The resistance could make LAB and yeast strains dominant microflora in the human gastrointestinal system; thus, competition against pathogen microorganisms can be provided more easily. Based on this circumstance, in the study, 10 LAB and 10 yeast strains were isolated from various dairy products, such as cheese, yoghurt, kefir, and cream. Samples were obtained from farmer markets and bazaars in Bursa, Turkey. As a part of this research, all isolated strains were identified and their ability of biofilm formation was detected with two different methods and compared with each other. The first goal of this research was to determine whether isolates have the potential for biofilm production, and the second was to compare the validity of two different methods, which are known as “Tube method” and “96-well plate-based method”. This study may offer an insight into developing a point of view about biofilm formation and its beneficial properties in LAB and yeast cultures used as a starter in the food industry.

Keywords: biofilm, dairy products, lactic acid bacteria, yeast

Procedia PDF Downloads 253

Authors: Muhammad Aown Sammar Raza

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Environmental stresses are one of the major reasons for poor crop yield across the globe. Among the various environmental stresses, drought stress is the most damaging one, especially in arid and semi-arid regions. Wheat is the major staple food of many countries of the world, which is badly affected by drought stress. In order to fulfill the dietary needs of increasing population with depleting water resources there is a need to adopt technologies which result in sufficient crop yield with less water consumption. One of them is partial root zone drying. Keeping in view these conditions, a wire house experiment was conducted at agronomic research area of University College of Agriculture and Environmental Sciences, The Islamia University Bahawalpur during 2015, to screen out the different wheat varieties for partial root zone drying (PRD). Five approved local wheat varieties (V1= Galaxy-2013, V2= Punjab-2011, V3 = Faisalabad-2008, V4 = Lasani-2008 and V5 = V.8200) and two irrigation levels (I1= control irrigation and I2 = PRD irrigation) with completely randomized design having four replications were used in the experiment. Among the varieties, Galaxy-2013 performed the best and attained maximum plant height, leaf area, stomatal conductance, photosynthesis, total sugars, proline contents and antioxidant enzymes activities and minimum values of growth and physiological parameters were recorded in variety V.8200. For irrigation levels, higher values of growth, physiological and water related parameters were recorded in control treatment (I1) except leaf water potential, osmotic potential, total sugars and proline contents. However, enzyme activities were higher under PRD treatment for all varieties. It was concluded that Galaxy-2013 is the most compatible and V.8200 is the most susceptible variety for PRD, respectively and more quality traits and enzymatic activities were recorded under PRD irrigation as compared to control treatment.

Keywords: antioxidant enzymes activities, osmolytes concentration, partial root zone drying, photosynthetic rate, water relations, wheat

Procedia PDF Downloads 236

Authors: Virve Pääkkönen, Heidi M. Cuffaro, Leo Tjäderhane

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Objectives: Odontoblasts are the outermost cell layer of dental pulp and form the dentin. Importance of bacterial products, e.g. lipoteichoic acid (LTA), a cell wall component of Gram-positive bacteria and lipopolysaccharide (LPS), a cell wall component of Gram-negative bacteria, have been indicated in the pathogenesis of pulpitis. Gram-positive bacteria are more prevalent in superficial carious lesion while the amount gram-negative is higher in the deep lesions. Objective of this study was to investigate the effect of these bacterial products on inflammatory response of pulp tissue. Interleukins (IL) were of special interest. Various ILs have been observed in the dentin-pulp complex of carious tooth in vivo. Methods: Tissue culture method was used for testing the effect of LTA and LPS on human odontoblasts. Enzymatic isolation technique was used to extract living odontoblasts for cell cultures. DNA microarray and quantitative PCR (qPCR) were used to characterize the changes in the expression profile of the tissue cultured odontoblasts. Laser microdissection was used to cut healthy and affected dentin and odontoblast layer directly under carious lesion for experiments. Cytokine array detecting 80 inflammatory cytokines was used to analyze the protein content of conditioned culture media as well as dentin and odontoblasts from the carious teeth. Results: LPS caused increased gene expression IL-1α, and -8 and decrease of IL-1β, 12 , -15 and -16 after 1h treatment, while after 24h treatment decrease of IL-8, -11 and 23 mRNAs was observed. LTA treatment caused cell death in the tissue cultured odontoblasts but in in the cell culture but not in cell culture. Cytokine array revealed at least 2-fold down-regulation of IL-1β, -10 and -12 in response to LPS treatment. Cytokine array of odontoblasts of carious teeth, as well as LPS-treated tissue-cultured odontoblasts, revealed increased protein amounts of IL-16, epidermal growth factor (EGF), angiogenin and IGFBP-1 as well as decreased amount of fractalkine. In carious dentin, increased amount of IL-1β, EGF and fractalkine was observed, as well as decreased level of GRO-1 and HGF. Conclusion: LPS caused marked changes in the expression of inflammatory cytokines in odontoblasts. Similar changes were observed in the odontoblasts cut directly under the carious lesion. These results help to shed light on the inflammatory processes happening during caries.

Keywords: inflammation, interleukin, lipoteichoic acid, odontoblasts

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Authors: Bengi Hakguder Taze, Sevcan Unluturk

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Şalak apricot (Prunus armeniaca L., cv. Şalak) is a specific variety grown in Igdir, Turkey. The fruit has distinctive properties distinguish it from other cultivars, such as its unique size, color, taste and higher water content. Drying is the widely used method for preservation of apricots. However, fresh consumption is preferred for Şalak apricot instead of drying due to its low dry matter content. Higher amounts of water in the structure and climacteric nature make the fruit sensitive against rapid quality loss during storage. Hence, alternative processing methods need to be introduced to extend the shelf life of the fresh produce. Mild heat (MH) treatment is of great interest as it can reduce the microbial load and inhibit enzymatic activities. Therefore, the aim of this study was to evaluate the impact of mild heat treatment on the natural microflora found on Şalak apricot surfaces and some physical quality parameters of the fruit, such as color and firmness. For this purpose, apricot samples were treated at different temperatures between 40 and 60 ℃ for different periods ranging between 10 to 60 min using a temperature controlled water bath. Natural flora on the fruit surfaces was examined using standard plating technique both before and after the treatment. Moreover, any changes in color and firmness of the fruit samples were also monitored. It was found that control samples were initially containing 7.5 ± 0.32 log CFU/g of total aerobic plate count (TAPC), 5.8±0.31 log CFU/g of yeast and mold count (YMC), and 5.17 ± 0.22 log CFU/g of coliforms. The highest log reductions in TAPC and YMC were observed as 3.87-log and 5.8-log after the treatments at 60 ℃ and 50 ℃, respectively. Nevertheless, the fruit lost its characteristic aroma at temperatures above 50 ℃. Furthermore, great color changes (ΔE ˃ 6) were observed and firmness of the apricot samples was reduced at these conditions. On the other hand, MH treatment at 41 ℃ for 10 min resulted in 1.6-log and 0.91-log reductions in TAPC and YMC, respectively, with slightly noticeable changes in color (ΔE ˂ 3). In conclusion, application of temperatures higher than 50 ℃ caused undesirable changes in physical quality of Şalak apricots. Although higher microbial reductions were achieved at those temperatures, temperatures between 40 and 50°C should be further investigated considering the fruit quality parameters. Another strategy may be the use of high temperatures for short time periods not exceeding 1-5 min. Besides all, MH treatment with UV-C light irradiation can be also considered as a hurdle strategy for better inactivation results.

Keywords: color, firmness, mild heat, natural flora, physical quality, şalak apricot

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Authors: Caroline M. Spagnol, Vera L. B. Isaac, Marcos A. Corrêa, Hérida R. N. Salgado

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Phenolic compounds are abundant in the Brazilian plant kingdom and they are part of a large and complex group of organic substances. Cinnamic acids are part of this group of organic compounds, and caffeic acid (CA) is one of its representatives. Antioxidants are compounds which act as free radical scavengers and, in other cases, such as metal chelators, both in the initiation stage and the propagation of oxidative process. The tyrosinase, polyphenol oxidase, is an enzyme that acts at various stages of melanin biosynthesis within the melanocytes and is considered a key molecule in this process. Some phenolic compounds exhibit inhibitory effects on melanogenesis by inhibiting the tyrosinase enzymatic activity and therefore has been the subject of studies. However, few studies have reported the effectiveness of these products and their safety. Objectives: To assess the inhibitory activity of tyrosinase, the antioxidant activity of CA and its cytotoxic potential. The method to evaluate the inhibitory activity of tyrosinase aims to assess the reduction transformation of L-dopa into dopaquinone reactions catalyzed by the enzyme. For evaluating the antioxidant activity was used the analytical methodology of DPPH radical inhibition. The cytotoxicity evaluation was carried out using the MTT method (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide), a colorimetric assay which determines the amount of insoluble violet crystals formed by the reduction of MTT in the mitochondria of living cells. Based on the results obtained during the study, CA has low activity as a depigmenting agent. However, it is a more potent antioxidant than ascorbic acid (AA), since a lower amount of CA is sufficient to inhibit 50% of DPPH radical. The results are promising since CA concentration that promoted 50% toxicity in HepG2 cells (IC50=781.8 μg/mL) is approximately 330 to 400 times greater than the concentration required to inhibit 50% of DPPH (IC50 DPPH= 2.39 μg/mL) and ABTS (IC50 ABTS= 1.96 μg/mL) radicals scavenging activity, respectively. The maximum concentration of caffeic acid tested (1140 mg /mL) did not reach 50% of cell death in HaCat cells. Thus, it was concluded that the caffeic acid does not cause toxicity in HepG2 and HaCat cells in the concentrations required to promote antioxidant activity in vitro, and it can be applied in topical products.

Keywords: caffeic acid, antioxidant, cytotoxicity, cosmetic

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Authors: F. A. M. Safwan, H. N. N. Dilrukshi, P. U. S. Peiris

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Less availability of high quality green forages leads to low productivity of national dairy herd of Sri Lanka. Growing grass and fodder to suit the production system is an efficient and economical solution for this problem. CO-3 is placed in a higher category, especially on tillering capacity, green forage yield, regeneration capacity, leaf to stem ratio, high crude protein content, resistance to pests and diseases and free from adverse factors along with other fodder varieties grown within the country. An experiment was designed to determine the effect of organic sheep manure, inorganic fertilizers and liquid sheep manure on growth, yield and nutritive value of CO-3. The study was consisted with three treatments; sheep manure (T1), recommended inorganic fertilizers (T2) and liquid sheep manure (T3) which was prepared using bucket fermentation method and each treatment was consisted with three replicates and those were assigned randomly. First harvest was obtained after 40 days of plant establishment and number of leaves (NL), leaf area (LA), tillering capacity (TC), fresh weight (FW) and dry weight (DW) were recorded and second harvest was obtained after 30 days of first harvest and same set of data were recorded. SPSS 16 software was used for data analysis. For proximate analysis AOAC, 2000 standard methods were used. Results revealed that the plants treated with T1 recorded highest NL, LA, TC, FW and DW and were statistically significant at first and second harvest of CO-3 (p˂ 0.05) and it was found that T1 was statistically significant from T2 and T3. Although T3 was recorded higher than the T2 in almost all growth parameters; it was not statistically significant (p ˃0.05). In addition, the crude protein content was recorded highest in T1 with the value of 18.33±1.61 and was lowest in T2 with the value of 10.82±1.14 and was statistically significant (p˂ 0.05). Apart from this, other proximate composition crude fiber, crude fat, ash, moisture content and dry matter were not statistically significant between treatments (p ˃0.05). In accordance with the results, it was found that the organic fertilizer is the best fertilizer for CO-3 in terms of growth parameters and crude protein content.

Keywords: fertilizer, growth parameters, Hybrid Napier CO-3, proximate composition

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Authors: Hoda M. Eid, Abir Nachar, Farah Thong, Gary Sweeney, Pierre S. Haddad

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Caffeic acid methyl ester (CAME) and caffeic ethyl esters (CAEE) were previously reported to potently stimulate glucose uptake in cultured C2C12 skeletal muscle cells via insulin-independent mechanisms involving the activation of adenosine monophosphate-activated protein kinase (AMPK). In the present study, we investigated the effect of the two compounds on the translocation of glucose transporter GLUT4 in L6 skeletal muscle cells. The cells were treated with the optimum non-toxic concentration (50 µM) of either CAME or CAEE for 18 h. Levels of GLUT4myc at the cell surface were measured by O-phenylenediamine dihydrochloride (OPD) assay. The effects of CAME and CAEE on GLUT1 and GLUT4 protein content were also measured by western immunoblot. Our results show that CAME and CAEE significantly increased glucose uptake, GLUT4 translocation and GLUT4 protein content. Furthermore, the effect of the two CA esters on two insulin-sensitive cell lines: H4IIE rat hepatoma and 3T3-L1 adipocytes were investigated. CAME and CAEE reduced the enzymatic activity of the key hepatic gluconeogenic enzyme glucose-6-phosphatase in a concentration-dependent manner. In addition, they exerted a concentration-dependent antiadipogenic effect on 3T3-L1 cells. Mitotic clonal expansion (MCE), a prerequisite for adipocytes differentiation was also concentration-dependently inhibited. The two compounds abrogated lipid droplet accumulation, blocked MCE and maintained cells in fibroblast-like state when applied at the maximum non-toxic concentration (100 µM). In addition, the expression of the early key adipogenic transcription factors CCAAT enhancer-binding protein beta (C/EBP-β) and the master regulator of adipogenesis peroxisome-proliferator-activated receptor gamma (PPAR-γ) were inhibited. We, therefore, conclude that CAME and CAEE exert pleiotropic benefits in several insulin-sensitive cell lines through insulin-independent mechanisms involving AMPK, hence they may treat obesity, diabetes and other metabolic diseases.

Keywords: type 2 diabetes mellitus, insulin resistance, GLUT4, Akt, AMPK.

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Authors: Josline Salib, Sayed El-Toumy, Abeer Salama, Enayat Omara, Emad Hassan

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Diabetic nephropathy is an important cause of morbidity and mortality and is now among the most common causes of end-stage renal failure (ESRF) in developed countries. Thus, the aim of the present study was to investigate the phenolic compounds content of Cichorium intybus aerial parts extracts as well as the therapeutic effects on diabetic nephropathy, oxidative stress, and anti-inflammatory by characterizing biochemical, histopathological changes and immunohistochemistry in an experimental diabetic rat model as compared with Amaryl. Ten known compounds of flavonoids, coumarins and phenolic acid derivatives were isolated from the C. intybus aqueous methanolic extract. Structures of the isolated compounds were established by chromatography, UV and 1D⁄2D 1H⁄ 13C spectroscopy. The aqueous methanol extract of C. intybus aerial parts was administered to Streptozotocin diabetes rats at doses (100 and 200 mg/kg) for 21 days. After treatment, blood glucose, serum insulin, urea, creatinine, and TNF-α were evaluated. Enzymatic scavengers including catalase (CAT), glutathione (GSH), malondialdehyde (MDA) and nitric oxide (NO) were determined to evaluate the oxidative status in the renal tissue. Diabetic rats treated with C. intybus extract showed a dose-dependent reduction of fasting blood glucose and kidney antioxidant status in comparison to the diabetic control group. The extract was able to enhance the antioxidant defenses of the kidney by increasing the reduced GSH and CAT content and decreasing MDA content in addition to significantly decreasing kidney nitric oxide content compared to diabetic control rats. Furthermore, the histopathological findings in C. intybus extract administered rats were observed at markedly lesser extent than the diabetic control group. Also, inducible nitric oxide synthase (iNOS) levels were decreased significantly after the administration of high-dose C. intybus extract in diabetic rats. Showing significant antihyperglycemic and antioxidant properties of C. intybus aerial parts extract, which is attributed to its polyphenolic content, may offer a potential source for the treatment of diabetes.

Keywords: antioxidant activity, anti-diabetic nephropathy, cichorium intybus aerial parts, phenolic compounds

Procedia PDF Downloads 113

Authors: Muhammad Ali Siddiquee, Md. Alauddin, Md. Omar Faruque, Zakir Hossain Howlader, Mohammad Asaduzzaman

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Rice bran (RB) and rice bran oil (RBO) are explored as prominent food components worldwide. In this study, fermented rice bran (FRB) was produced by employing edible gram-positive bacteria (Lactobacillus acidophilus, Lactobacillus bulgaricus, and Bifidobacterium bifidum) at 125 x 10⁵ spore g⁻¹ of rice bran, and investigated to evaluate nutritional quality. The crude rice bran oil (CRBO) was extracted from RB, and its quality was also investigated compared to market-available rice bran oil (MRBO) in Bangladesh. We found that fermentation of rice bran with lactic acid bacteria increased total proteins (29.52%), fat (5.38%), ash (48.47%), crude fiber (38.96%), and moisture (61.04%) and reduced the carbohydrate content (36.61%). We also found that essential amino acids (methionine, tryptophan, threonine, valine, leucine, lysine, histidine, and phenylalanine) and non-essential amino acids (alanine, aspartate, glycine, glutamine, proline, serine, and tyrosine) were increased in FRB except methionine and proline. Moreover, total phenolic content, tannin content, flavonoid content, and antioxidant activity were increased in FRB. The RBO analysis showed that γ-oryzanol content (10.00mg/g) was found in CRBO compared to MRBO (ranging from 7.40 to 12.70 mg/g) and Vitamin-E content 0.20% was found higher in CRBO compared to MRBO (ranging 0.097 to 0.12%). The total saturated (25.16%) and total unsaturated fatty acids (74.44%) were found in CRBO, whereas MRBO contained total saturated (22.08 to 24.13%) and total unsaturated fatty acids (71.91 to 83.29%), respectively. The physiochemical parameters were found satisfactory in all samples except acid value and peroxide value higher in CRBO. Finally, animal experiments showed that FRB and CRBO reduce the body weight, glucose, and lipid profile in high-fat diet-induced animal models. Thus, FRB and RBO could be value-added food supplements for human health.

Keywords: fermented rice bran, crude rice bran oil, amino acids, proximate composition, gamma-oryzanol, fatty acids, heavy metals, physiochemical parameters

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Authors: Evgeniya Y. Yuzbasheva, Tigran V. Yuzbashev, Natalia I. Perkovskaya, Elizaveta B. Mostova

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Cell-surface display of lipase is of great interest as it has many applications in the field of biotechnology owing to its unique advantages: simplified product purification, and cost-effective downstream processing. One promising area of application for whole-cell biocatalysts with surface displayed lipase is biodiesel synthesis. Biodiesel is biodegradable, renewable, and nontoxic alternative fuel for diesel engines. Although the alkaline catalysis method has been widely used for biodiesel production, it has a number of limitations, such as rigorous feedstock specifications, complicated downstream processes, including removal of inorganic salts from the product, recovery of the salt-containing by-product glycerol, and treatment of alkaline wastewater. Enzymatic synthesis of biodiesel can overcome these drawbacks. In this study, Lip2p lipase was displayed on Yarrowia lipolytica cells via C- and N-terminal fusion variant. The active site of lipase is located near the C-terminus, therefore to prevent the activity loosing the insertion of glycine-serine linker between Lip2p and C-domains was performed. The hydrolytic activity of the displayed lipase reached 12,000–18,000 U/g of dry weight. However, leakage of enzyme from the cell wall was observed. In case of C-terminal fusion variant, the leakage was occurred due to the proteolytic cleavage within the linker peptide. In case of N-terminal fusion variant, the leaking enzyme was presented as three proteins, one of which corresponded to the whole hybrid protein. The calculated number of recombinant enzyme displayed on the cell surface is approximately 6–9 × 105 molecules per cell, which is close to the theoretical maximum (2 × 106 molecules/cell). Thus, we attribute the enzyme leakage to the limited space available on the cell surface. Nevertheless, cell-bound lipase exhibited greater stability to short-term and long-term temperature treatment than the native enzyme. It retained 74% of original activity at 60°C for 5 min of incubation, and 83% of original activity after incubation at 50°C during 5 h. Cell-bound lipase had also higher stability in organic solvents and detergents. The developed whole-cell biocatalyst was used for recycling biodiesel synthesis. Two repeated cycles of methanolysis yielded 84.1–% and 71.0–% methyl esters after 33–h and 45–h reactions, respectively.

Keywords: biodiesel, cell-surface display, lipase, whole-cell biocatalyst

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Authors: Milda Nainyte, Viktoras Masevicius

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Biological methylation is a methyl group transfer from S-adenosyl-L-methionine (AdoMet) onto N-, C-, O- or S-nucleophiles in DNA, RNA, proteins or small biomolecules. The reaction is catalyzed by enzymes called AdoMet-dependent methyltransferases (MTases), which represent more than 3 % of the proteins in the cell. As a general mechanism, the methyl group from AdoMet replaces a hydrogen atom of nucleophilic center producing methylated DNA and S-adenosyl-L-homocysteine (AdoHcy). Recently, DNA methyltransferases have been used for the sequence-specific, covalent labeling of biopolymers. Two types of MTase catalyzed labeling of biopolymers are known, referred as two-step and one-step. During two-step labeling, an alkylating fragment is transferred onto DNA in a sequence-specific manner and then the reporter group, such as biotin, is attached for selective visualization using suitable chemistries of coupling. This approach of labeling is quite difficult and the chemical hitching does not always proceed at 100 %, but in the second step the variety of reporter groups can be selected and that gives the flexibility for this labeling method. In the one-step labeling, AdoMet analog is designed with the reporter group already attached to the functional group. Thus, the one-step labeling method would be more comfortable tool for labeling of biopolymers in order to prevent additional chemical reactions and selection of reaction conditions. Also, time costs would be reduced. However, effective AdoMet analog appropriate for one-step labeling of biopolymers and containing cleavable bond, required for reduction of PCR interferation, is still not known. To expand the practical utility of this important enzymatic reaction, cofactors with activated sulfonium-bound side-chains have been produced and can serve as surrogate cofactors for a variety of wild-type and mutant DNA and RNA MTases enabling covalent attachment of these chains to their target sites in DNA, RNA or proteins (the approach named methyltransferase-directed Transfer of Activated Groups, mTAG). Compounds containing hex-2-yn-1-yl moiety has proved to be efficient alkylating agents for labeling of DNA. Herein we describe synthetic procedures for the preparation of N-biotinoyl-N’-(pent-4-ynoyl)cystamine starting from the coupling of cystamine with pentynoic acid and finally attaching the biotin as a reporter group. The synthesis of the first AdoMet based cofactor containing a cleavable reporter group and appropriate for one-step labeling was developed.

Keywords: adoMet analogs, DNA alkylation, cofactor, methyltransferases

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Authors: Aktar-Uz-Zaman, M. Tuhina-Khatun, Mohamed Hanafi Musa

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Three rust diseases: leaf (brown) rust caused by Puccinia triticina Eriks, stripe (yellow) rust caused by Puccinia striiformis West, and stem (black) rust caused by Puccinia graminis f. sp. tritici are economically important diseases of wheat in world wide. Yield loss due to leaf rust is 40% in susceptible cultivars. Yield losses caused by the stem rust pathogens in the mid of 20 century reached 20-30% in Eastern and Central Europe and the most virulent stem rust race Ug99 emerged first in Uganda and after that in Kenya, Ethiopia, Yemen, in the Middle East and South Asia. Yield losses were estimated up to 100%, whereas, up to 80% have been reported in Kenya during 1999. In case of stripe rust, severity level has been recorded 60% - 70% as compared to 100% severity of susceptible check in disease screening nurseries in Kenya. Improvement of resistant varieties or cultivars is the sustainable, economical and environmentally friendly approaches for increasing the global wheat production to suppress the rust diseases. More than 68 leaf rust, 49 stripe rust and 53 stem rust resistance genes have been identified in the global wheat cultivars or varieties using different molecular breeding approaches. Among these, Lr1, Lr9, Lr10, Lr19, Lr21, Lr24, Lr25, Lr28, Lr29, Lr34, Lr35, Lr37, Lr39, Lr47, Lr51, Lr3bg, Lr18, Lr40, Lr46, and Lr50 leaf rust resistance genes have been identified by using molecular, enzymatic and microsatellite markers from African, Asian, European cultivars of hexaploid wheat (Triticum aestivum), durum wheat and diploid wheat species. These genes are located on 20, of the 21 chromosomes of hexaploid wheat. Similarly, Sr1, Sr2, Sr24, and Sr3, Sr31 stem rust resistance genes have been recognized from wheat cultivars of Pakistan, India, Kenya, and Uganda etc. A race of P. striiformis (stripe rust) Yr9, Yr18, and Yr29 was first observed in East Africa, Italy, Pakistan and India wheat cultivars. These stripe rust resistance genes are located on chromosomes 1BL, 4BL, 6AL, 3BS and 6BL in bread wheat cultivars. All these identified resistant genes could be used for notable improvement of susceptible wheat cultivars in the future.

Keywords: hexaploid wheat, resistance genes, rust disease, triticum aestivum

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Authors: Zeineb Baatout, Safa Teka, Nejmeddine Jaballah, Nawfel Sakly, Xiaonan Sun, Mustapha Majdoub

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Currently, the economic interests linked to the development of bio-based materials make biomass one of the most interesting areas for science development. We are interested in the β-cyclodextrin (β-CD), one of the popular bio-sourced macromolecule, produced from the starch via enzymatic conversion. It is a cyclic oligosaccharide formed by the association of seven glucose units. It presents a rigid conical and amphiphilic structure with hydrophilic exterior, allowing it to be water-soluble. It has also a hydrophobic interior enabling the formation of inclusion complexes, which support its application for the elaboration of electrochemical and optical sensors. Nevertheless, the solubility of β-CD in water makes its use as sensitive layer limit and difficult due to their instability in aqueous media. To overcome this limitation, we chose to precede by modification of the hydroxyl groups to obtain hydrophobic derivatives which lead to water-stable sensing layers. Hence, a series of benzylated β-CDs were synthesized in basic aqueous media in one pot. This work reports the synthesis of a new family of substituted amphiphilic β-CDs using a green methodology. The obtained β-CDs showed different degree of substitution (DS) between 0.85 and 2.03. These organic macromolecular materials were soluble in common organic volatile solvents, and their structures were investigated by NMR, FT-IR and MALDI-TOF spectroscopies. Thermal analysis showed a correlation between the thermal properties of these derivatives and the benzylation degree. The surface properties of the thin films based on the benzylated β-CDs were characterized by contact angle measurements and atomic force microscopy (AFM). These organic materials were investigated as sensitive layers, deposited on quartz crystal microbalance (QCM) gravimetric transducer, for humidity sensor at room temperature. The results showed that the performances of the prepared sensors are greatly influenced by the benzylation degree of β-CD. The partially modified β-CD (DS=1) shows linear response with best sensitivity, good reproducibility, low hysteresis, fast response time (15s) and recovery time (17s) at higher relative humidity levels (RH) between 11% and 98% in room temperature.

Keywords: β-cyclodextrin, green synthesis, humidity sensor, quartz crystal microbalance

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Authors: Dibakar Chandra Deka, Anamika Kalita Deka

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Fermented alcoholic beverage production has its own stand among the tribal communities of North-East India. This biological oxidation method is followed by Ahom, Dimasa, Nishi, Miri, Bodo, Rabha tribes of this region. Bodo tribes among them not only prepare fermented alcoholic beverage but also store it for various time periods like 3 months, 6 months, 9 months, 12 months and 15 months etc. They prepare alcoholic beverage Jou (rice beer) following the fermentation of Oryza sativa with traditional yeast culture Amao. Saccharomyces cerevisiae is the main domain strain present in Amao. Dongphangrakep (Scoparia dulcis), Mwkhna (Clerodendrum viscosum), Thalir (Musa balbisina) and Khantal Bilai (Ananas cosmos) are the main plants used for Amao preparation. The stored Jou is known as Joufinai. They store the fermented mixture (rice and Amao) in anaerobic conditions for the preparation of Joufinai. We observed a successive increase in alcohol content from 3 months of storage period with 11.79 ± 0.010 (%, v/v) to 15.48 ± 0.070 (%, v/v) at 15 months of storage by a simple, reproducible and solution based colorimetric method. A positive linear correlation was also observed between pH and ethanol content with storage having correlation coefficient 0.981. Here, we optimised the detection of change in constituents of Joufinai during storage using reverse phase HPLC method. We found acetone, ethanol, acetic acid, glycerol as main constituents present in Joufinai. A very good correlation was observed from 3 months to 15 months of storage periods with its constituents. Increase in glycerol content was also detected with storage periods and hence Joufinai can be use as a precursor of above stated compounds. We also observed antioxidant activities increase from 0.056 ±2.80 mg/mL for 3 months old to 0.078± 5.33 mg/mL (in ascorbic acid equivalents) for 15 month old beverage by DPPH radical scavenging method. Therefore, we aimed for scientific validation of storage procedure used by Bodos in Joufinai production and to convert the Bodos’ traditional alcoholic beverage to a commercial commodity through our study.

Keywords: Amao, correlation, beverage, joufinai

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Authors: Amr Saeb, Khalid Al-Rubeaan, Mohamed Abouelhoda, Manojkumar Selvaraju, Hamsa Tayeb

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Background: P. mirabilis is a common uropathogenic bacterium that can cause major complications in patients with long-standing indwelling catheters or patients with urinary tract anomalies. In addition, P. mirabilis is a common cause of chronic osteomyelitis in diabetic foot ulcer (DFU) patients. Methodology: P. mirabilis SCDR1 was isolated from a diabetic ulcer patient. We examined P. mirabilis SCDR1 levels of resistance against nano-silver colloids, the commercial nano-silver and silver containing bandages and commonly used antibiotics. We utilized next generation sequencing techniques (NGS), bioinformatics, phylogenetic analysis and pathogenomics in the identification and characterization of the infectious pathogen. Results: P. mirabilis SCDR1 is a multi-drug resistant isolate that also showed high levels of resistance against nano-silver colloids, nano-silver chitosan composite and the commercially available nano-silver and silver bandages. The P. mirabilis-SCDR1 genome size is 3,815,621 bp with G+C content of 38.44%. P. mirabilis-SCDR1 genome contains a total of 3,533 genes, 3,414 coding DNA sequence genes, 11, 10, 18 rRNAs (5S, 16S, and 23S), and 76 tRNAs. Our isolate contains all the required pathogenicity and virulence factors to establish a successful infection. P. mirabilis SCDR1 isolate is a potential virulent pathogen that despite its original isolation site, wound, it can establish kidney infection and its associated complications. P. mirabilis SCDR1 contains several mechanisms for antibiotics and metals resistance including, biofilm formation, swarming mobility, efflux systems, and enzymatic detoxification. Conclusion: P. mirabilis SCDR1 is the spontaneous nano-silver resistant bacterial strain. P. mirabilis SCDR1 strain contains all reported pathogenic and virulence factors characteristic for the species. In addition, it possesses several mechanisms that may lead to the observed nano-silver resistance.

Keywords: Proteus mirabilis, multi-drug resistance, silver nanoparticles, resistance, next generation sequencing techniques, genome analysis, bioinformatics, phylogeny, pathogenomics, diabetic foot ulcer, xenobiotics, multidrug resistance efflux, biofilm formation, swarming mobility, resistome, glutathione S-transferase, copper/silver efflux system, altruism

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Authors: Yogesh Rai, Saurabh Singh, Sanjay Pandey, Dhananjay K. Sah, B. G. Roy, B. S. Dwarakanath, Anant N. Bhatt

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One of the most common signatures of highly malignant gliomas is their capacity to metabolize more glucose to lactic acid than normal brain tissues, even under normoxic conditions (Warburg effect), indicating that aerobic glycolysis is constitutively upregulated through stable genetic or epigenetic changes. However, oxidative phosphorylation (OxPhos) is also required to maintain the mitochondrial membrane potential for tumor cell survival. In the process of tumorigenesis, tumor cells during fastest growth rate exhibit both high glycolytic and high OxPhos. Therefore, metabolically reprogrammed cancer cells with combination of both aerobic glycolysis and altered OxPhos develop a robust metabolic phenotype, which confers a selective growth advantage. In our study, we grew the high glycolytic BMG-1 (glioma) cells with continuous exposure of mitochondrial uncoupler 2, 4, dinitro phenol (DNP) for 10 passages to obtain a phenotype of high glycolysis with enhanced altered OxPhos. We found that OxPhos modified BMG (OPMBMG) cells has similar growth rate and cell cycle distribution but high mitochondrial mass and functional enzymatic activity than parental cells. In in-vitro studies, OPMBMG cells showed enhanced invasion, proliferation and migration properties. Moreover, it also showed enhanced angiogenesis in matrigel plug assay. Xenografted tumors from OPMBMG cells showed reduced latent period, faster growth rate and nearly five folds reduction in the tumor take in nude mice compared to BMG-1 cells, suggesting that robust metabolic phenotype facilitates tumor formation and growth. OPMBMG cells which were found radio-resistant, showed enhanced radio-sensitization by 2-DG as compared to the parental BMG-1 cells. This study suggests that metabolic reprogramming in cancer cells enhances the potential of migration, invasion and proliferation. It also strengthens the cancer cells to escape the death processes, conferring resistance to therapeutic modalities. Our data also suggest that combining metabolic inhibitors like 2-DG with conventional therapeutic modalities can sensitize such metabolically aggressive cancer cells more than the therapies alone.

Keywords: 2-DG, BMG, DNP, OPM-BMG

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Authors: Karol Rodriguez-Peña, Martha L. Macias-Rubalcava, Leticia Rocha-Zavaleta, Sergio Sanchez

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A bioassay-guided study for anti-cancer compounds from endophytes of the Mexican medicinal plant Amphipteryygium adstringens resulted in the isolation of a streptomycete capable of producing a group of compounds with high cytotoxic activity. Microorganisms from surface sterilized samples of various sections of the plant were isolated and all the actinomycetes found were evaluated for their potential to produce compounds with cytotoxic activity against cancer cell lines MCF7 (breast cancer) and HeLa (cervical cancer) as well as the non-tumoural cell line HaCaT (keratinocyte). The most active microorganism was picked for further evaluation. The identification of the microorganism was carried out by 16S rDNA gene sequencing, finding the closest proximity to Streptomyces scabrisporus, but with the additional characteristic that the strain isolated in this study was capable of producing colorful compounds never described for this species. Crude extracts of dichloromethane and ethyl acetate showed IC50 values of 0.29 and 0.96 μg/mL for MCF7, 0.51 and 1.98 μg/mL for HeLa and 0.96 and 2.7 μg/mL for HaCaT. Scaling the fermentation to 10 L in a bioreactor generated 1 g of total crude extract, which was fractionated by silica gel open column to yield 14 fractions. Nine of the fractions showed cytotoxic activity. Fraction 4 was chosen for subsequent purification because of its high activity against cancerous cell lines, lower activity against keratinocytes. HPLC-UV-MS/ESI was used for the evaluation of this fraction, finding at least 10 different compounds with high values of m/z (≈588). Purification of the compounds was carried out by preparative thin-layer chromatography. The prevalent compound was Steffimycin B, a molecule known for its antibiotic and cytotoxic activities and also for its low solubility in aqueous solutions. Along with steffimycin B, another five compounds belonging to the steffimycin family were isolated and at this moment their structures are being elucidated, some of which display better solubility in water: an attractive property for the pharmaceutical industry. As a conclusion to this study, the isolation of endophytes resulted in the discovery of a strain capable of producing compounds with high cytotoxic activity that need to be studied for their possible utilization.

Keywords: amphipterygium adstringens, cytotoxicity, streptomyces scabrisporus, steffimycin

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Authors: G. F. Okunade, M. O. Lawal, R. E. Uwadiae, D. Portnoy

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The diversity, physico-chemical, biochemical and genomics assessment of Macrofauna species of Ologe and Badagry Lagoons were carried out between August 2016 and July 2018. The concentrations of Fe, Zn, Mn, Cd, Cr, and Pb in water were determined by Atomic Absorption Spectrophotometer (AAS). Particle size distribution was determined with wet-sieving and sedimentation using hydrometer method. Genomics analyses were carried using 25 P. fusca (quadriseriata) and 25 P.fusca from each lagoon due to abundance in both lagoons all through the two years of collection. DNA was isolated from each sample using the Mag-Bind Blood and Tissue DNA HD 96 kit; a method designed to isolate high quality. The biochemical characteristics were analysed in the dominanat species (P.aurita and T. fuscatus) using ELISA kits. Physico-chemical parameters such as pH, total dissolved solids, dissolved oxygen, conductivity and TDS were analysed using APHA standard protocols. The Physico-chemical parameters of the water quality recorded with mean values of 32.46 ± 0.66mg/L and 41.93 ± 0.65 for COD, 27.28 ± 0.97 and 34.82 ± 0.1 mg/L for BOD, 0.04 ± 4.71 mg/L for DO, 6.65 and 6.58 for pH in Ologe and Badagry lagoons with significant variations (p ≤ 0.05) across seasons. The mean and standard deviation of salinity for Ologe and Badagry Lagoons ranged from 0.43 ± 0.30 to 0.27 ± 0.09. A total of 4210 species belonging to a phylum, two classes, four families and a total of 2008 species in Ologe lagoon while a phylum, two classes, 5 families and a total of 2202 species in Badagry lagoon. The percentage composition of the classes at Ologe lagoon had 99% gastropod and 1% bivalve, while Gastropod contributed 98.91% and bivalve 1.09% in Badagry lagoon. Particle size was distributed in 0.002mm to 2.00mm, particle size distribution in Ologe lagoon recorded 0.83% gravels, 97.83% sand, and 1.33% silt particles while Badagry lagoon recorded 7.43% sand, 24.71% silt, and 67.86% clay particles hence, the excessive dredging activities going on in the lagoon. Maximum percentage of sand (100%) was seen in station 6 in Ologe lagoon while the minimum (96%) was found in station 1. P. aurita (Ologe Lagoon) and T. fuscastus (Badagry Lagoon) were the most abundant benthic species in which both contributed 61.05% and 64.35%, respectively. The enzymatic activities of P. aurita observed with mean values of 21.03 mg/dl for AST, 10.33 mg/dl for ALP, 82.16 mg/dl for ALT and 73.06 mg/dl for CHO in Ologe Lagoon While T. fuscatus observed mean values of Badagry Lagoon) recorded mean values 29.76 mg/dl, ALP with 11.69mg/L, ALT with 140.58 mg/dl and CHO with 45.98 mg/dl. There were significant variations (P < 0.05) in AST and CHO levels of activities in the muscles of the species.

Keywords: benthos, biochemical responses, genomics, metals, particle size

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Authors: Seunghwan Seo, Woo-Seok Lee, Ji-Sun Shin, Young Kyoung Rhee, Chang-Won Cho, Hee-Do Hong, Kyung-Tae Lee

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Doenjang, known as traditional Korean food, is product of a natural mixed fermentation process carried out by lactic acid bacteria (LAB). Lactobacillus sakei K040706 (K040706) has been accepted as the most populous LAB in over ripened doenjang. Recently, we reported the immunostimulatory effects of K040706 in RAW 264.7 macrophages and in a cyclophosphamide-induced mouse model. In this study, we investigated the ameliorative effects of K040706 in a dextran sulfate sodium (DSS)-induced colitis mouse model. We induced colitis using DSS in 5-week-ICR mice over 14 days with or without 0.1, 1 g/kg/day K040706 orally. The body weight, stool consistency, and gross bleeding were recorded for determination of the disease activity index (DAI). At the end of treatment, animals were sacrificed and colonic tissues were collected and subjected to histological experiments and myeloperoxidase (MPO) accumulation, cytokine determination, qRT-PCR and Western blot analysis. Results showed that K040706 significantly attenuated DSS-induced DAI score, shortening of colon length, enlargement of spleen and immune cell infiltrations into colonic tissues. Histological examinations indicated that K040706 suppressed edema, mucosal damage, and the loss of crypts induced by DSS. These results were correlated with the restoration of tight junction protein expression, such as, ZO-1 and occludin in K040706-treated mice. Moreover, K040706 reduced the abnormal secretions and mRNA expressions of pro-inflammatory mediators, such as nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). DSS-induced mRNA expression of intercellular adhesion molecule (ICAM) and vascular cell adhesion molecule (VCAM) in colonic tissues was also downregulated by K040706 treatment. Furthermore, K040706 suppressed the protein and mRNA expression of toll-like receptor 4 (TLR4) and phosphorylation of NF-κB and signal transducer and activator of transcription 3 (STAT3). These results suggest that K040706 has an anti-colitic effect by inhibition of intestinal inflammatory responses in DSS-induced colitic mice.

Keywords: Lactobacillus sakei, NF-κB, STAT3, ulcerative colitis

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Authors: Bahareh Sarmadi, Paul A. Kilmartin, Leandro D. Araújo, Brandt P. Bastow

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Aim: Sauvignon blanc is the most substantial variety cultivated in almost 62% of all producing vineyards of New Zealand. The popularity of New Zealand Sauvignon blanc is due to its unique taste. It is the most famous wine characterized by its aroma profile derived from mercaptans. 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl acetate (3MHA) are two of the most important volatile mercaptans found in Sauvignon blanc wines. “Viticultural” and “Enological” factors such as machine-harvesting, the most common harvesting practice used in New Zealand, can be among the reasons for this distinct flavor. Elemental sulfur is commonly sprayed in the fields to protect berries against powdery mildew. Although it is not the only source of sulfur, this practice creates a source of elemental sulfur that can be transferred into the must and eventually into wines. Despite the clear effects of residual elemental sulfur present in the must on the quality and aroma of the final wines, its measurement before harvest or fermentation is not a regular practice in the wineries. This can be due to the lack of accessible and applicable methods for the equipment at most commercial wineries. This study aims to establish a relationship between the number and frequency of elemental sulfur applications and the concentration of polyfunctional mercaptans in the final wines. Methods: An apparatus was designed to reduce elemental sulfur to sulfide, then an ion-selective electrode to measure sulfide concentration. During harvest 2022, we explored a wider range of residual elemental sulfur levels than what typically applies in the vineyards. This has been done through later manual elemental sulfur applications in the vineyard. Additional sulfur applications were made 20, 10 and 5 days prior to harvesting the treated grapes, covering long and short pre-harvest intervals (PHI). The grapes were processed into juice and fermented into wine; then, they were analyzed to find the correlation between polyfunctional mercaptans concentrations in the wines and residual elemental sulfur in the juice samples. Results: The research showed that higher 3MH/3MHA was formed when elemental sulfur was applied more frequent in the vineyards and supported the proposed pathway in which elemental sulfur is a source of 3MH formation in wines.

Keywords: sauvignon blanc, elemental sulfur, polyfunctional mercaptans, varietal thiols

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Authors: Mario Perez-Won, Roberto Lemus-Mondaca, Constanza Olivares-Rivera, Fernanda Marin-Monardez

Abstract:

This research presents the simultaneous application of high hydrostatic pressure (HHP) and osmotic dehydration (DO) as a pretreatment to hot –air drying of jumbo squid (Dosidicus gigas) cubes. The drying time was reduced to 2 hours at 60ºC and 5 hours at 40°C as compared to the jumbo squid samples untreated. This one was due to osmotic pressure under high-pressure treatment where increased salt saturation what caused an increasing water loss. Thus, a more reduced time during convective drying was reached, and so water effective diffusion in drying would play an important role in this research. Different working conditions such as pressure (350-550 MPa), pressure time (5-10 min), salt concentration, NaCl (10 y 15%) and drying temperature (40-60ºC) were optimized according to kinetic parameters of each mathematical model. The models used for drying experimental curves were those corresponding to Weibull, Page and Logarithmic models, however, the latest one was the best fitted to the experimental data. The values for water effective diffusivity varied from 4.82 to 6.59x10-9 m2/s for the 16 curves (DO+HHP) whereas the control samples obtained a value of 1.76 and 5.16×10-9 m2/s, for 40 and 60°C, respectively. On the other hand, quality characteristics such as color, texture, non-enzymatic browning, water holding capacity (WHC) and rehydration capacity (RC) were assessed. The L* (lightness) color parameter increased, however, b * (yellowish) and a* (reddish) parameters decreased for the DO+HHP treated samples, indicating treatment prevents sample browning. The texture parameters such as hardness and elasticity decreased, but chewiness increased with treatment, which resulted in a product with a higher tenderness and less firmness compared to the untreated sample. Finally, WHC and RC values of the most treatments increased owing to a minor damage in tissue cellular compared to untreated samples. Therefore, a knowledge regarding to the drying kinetic as well as quality characteristics of dried jumbo squid samples subjected to a pretreatment of osmotic dehydration under high hydrostatic pressure is extremely important to an industrial level so that the drying process can be successful at different pretreatment conditions and/or variable processes.

Keywords: diffusion coefficient, drying process, high pressure, jumbo squid, modelling, quality aspects

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