Search results for: lactic acid bacteria

Authors: Reenu Yadav, Abhay Asthana, S. K. Yadav

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Purpose: The aim of the present work was to develop an oral gel for brushing with an antimicrobial activity which will cure/protect from various periodontal diseases such as periodontitis, gingivitis, and pyorrhea. Methods: Plant materials procured from local suppliers, extracted and standardized. Screening of antimicrobial activity was carried out with the help of disk diffusion method. The gel was formulated by dried extracts of Beautea monosperma and Cordia obliquus. Gels were evaluated on various parameters and standardization of the formulation was performed. The release of drugs was studied in pH 6.8 using a mastication device.Total phenolic and flavonoid contents were estimated by folin-Ciocalteu and aluminium chloride method, and stability studies were performed (40°C and RH 75% ± 5% for 90 days) to assess the effect of temperature and humidity on the concentration of phenolic and flavonoid contents. The results of accelerated stability conditions were compared with that of samples kept at controlled conditions (RT). The control samples were kept at room temperature (25°C, 35% RH for 180 days). Results: Results are encouraging; extracts possess significant antimicrobial activity at very low concentration (15µg/disc, 20µg/disc and 15 µg/ disc) on oral pathogenic bacteria. The formulation has optimal characteristics, as well as has a pleasant appearance, fragrance, texture, and taste, is highly acceptable by the volunteers. The diffusion coefficient values ranged from 0.6655 to 0.9164. Since the R values of korsmayer papas were close to 1, Drug release from formulation follows matrix diffusion kinetics. Hence, diffusion was the mechanism of the drug release. Formulation follows non-Fickian transport mechanism. Most Formulations released 50 % of their contents within 25-30 minutes. Results obtained from the accelerated stability studies are indicative of a slight reduction in flavonoids and phenolic contents with time on long time storage. When measured degradation under ambient conditions, degradation was significantly lower than in accelerated stability study. Conclusion: Plant extracts possess compounds with antimicrobial properties can be used as. Developed formulation will cure/protect from various periodontal diseases. Further development and evaluations oral gel including the isolated compounds on the commercial scale and their clinical and toxicological studies are the future challenges.

Keywords: herbal gel, dental care, ambient conditions, commercial scale

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Authors: Shadia Al-Bahlani, Khadija Al-Bulushi, Zuweina Al-Hadidi, Buthaina Al-Dhahl, Nadia Al-Abri

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Breast cancer is the most common cancer in women worldwide. Triple-Negative Breast Cancer (TNBC) is an aggressive type of breast cancer, which is defined by the absence of Estrogen (ER), Progesterone (PR) and human epidermal growth factor (Her-2) receptors. The calpain system plays an important role in many cellular processes including apoptosis, necrosis, cell signaling and proliferation. However, the role of calpain in cisplatin (CDDP)-induced apoptosis in TNBC cells is not fully understood. Here, TNBC (MDA-MB231) cells were treated with different concentration of CDDP (0, 20 & 40 µM) and calpain activation and apoptosis were measured by western blot and Hoechst Stain respectively. In addition, calpain modulation by either activation and/or inhibition and its effect on CDDP-induced apoptosis were assessed by the same above approaches. Our findings showed that CDDP induced endoplasmic reticulum stress and thus Calcium release and subsequently activate calpain α-fodrin cleavage indicated by the increase in GRP78 and Calmodulin protein expression and respectively in MDA-MB231 cells. It also induced apoptosis as measured by Hoechst stain and caspase-12 cleavage. Calpain activation by both Cyclopiazonic acid and Thapsigargin showed similar effect and enhanced the sensitivity of these cells to CDDP treatment. On the other hand, calpain inhibition by either specific siRNA and/or exogenous inhibitor (Calpeptin) had an adverse effect where it attenuated calpain activation and thus CDDP- induced apoptosis in these cells. Altogether, these findings suggested that calpain activation play an essential role in sensitizing the TNBC cells to CDDP-induced apoptosis. This might lead to the discovery of novel treatment to over this aggressive type of breast cancer.

Keywords: calpain, cisplatin, apoptosis, breast cancer

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Authors: Khin Thanda Win, Chunying Zhang, Sanghyeob Lee

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Mildew resistance locus o (Mlo), a plant-specific gene family with seven-transmembrane (TM), plays an important role in plant resistance to powdery mildew (PM). PM caused by Podosphaera xanthii is a widespread plant disease and probably represents the major fungal threat for many Cucurbits. The recent Cucurbita maxima genome sequence data provides an opportunity to identify and characterize the MLO gene family in this species. Total twenty genes (designated CmaMLO1 through CmaMLO20) have been identified by using an in silico cloning method with the MLO gene sequences of Cucumis sativus, Cucumis melo, Citrullus lanatus and Cucurbita pepo as probes. These CmaMLOs were evenly distributed on 15 chromosomes of 20 C. maxima chromosomes without any obvious clustering. Multiple sequence alignment showed that the common structural features of MLO gene family, such as TM domains, a calmodulin-binding domain and 30 important amino acid residues for MLO function, were well conserved. Phylogenetic analysis of the CmaMLO genes and other plant species reveals seven different clades (I through VII) and only clade IV is specific to monocots (rice, barley, and wheat). Phylogenetic and structural analyses provided preliminary evidence that five genes belonged to clade V could be the susceptibility genes which may play the importance role in PM resistance. This study is the first comprehensive report on MLO genes in C. maxima to our knowledge. These findings will facilitate the functional analysis of the MLOs related to PM susceptibility and are valuable resources for the development of disease resistance in pumpkin.

Keywords: Mildew resistance locus o (Mlo), powdery mildew, phylogenetic relationship, susceptibility genes

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Authors: Maliheh Raji, Hossein Abolghasemi, Jaber Safdari, Ali Kargari

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Dysprosium is a rare earth element which is essential for many growing high-technology applications. Dysprosium along with neodymium plays a significant role in different applications such as metal halide lamps, permanent magnets, and nuclear reactor control rods preparation. The purification and separation of rare earth elements are challenging because of their similar chemical and physical properties. Among the various methods, membrane processes provide many advantages over the conventional separation processes such as ion exchange and solvent extraction. In this work, selective extraction and separation of dysprosium from aqueous solutions containing an equimolar mixture of dysprosium and neodymium by emulsion liquid membrane (ELM) was investigated. The organic membrane phase of the ELM was a nanofluid consisting of multiwalled carbon nanotubes (MWCNT), Span80 as surfactant, Cyanex 272 as carrier, kerosene as base fluid, and nitric acid solution as internal aqueous phase. Factors affecting separation of dysprosium such as carrier concentration, MWCNT concentration, feed phase pH and stripping phase concentration were analyzed using Taguchi method. Optimal experimental condition was obtained using analysis of variance (ANOVA) after 10 min extraction. Based on the results, using MWCNT nanofluid in ELM process leads to increase the extraction due to higher stability of membrane and mass transfer enhancement and separation factor of 6 for dysprosium over neodymium can be achieved under the optimum conditions. Additionally, demulsification process was successfully performed and the membrane phase reused effectively in the optimum condition.

Keywords: emulsion liquid membrane, MWCNT nanofluid, separation, Taguchi method

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Authors: Thidarat Imyen, Paisan Kongkachuichay

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Cu-Zn/core-shell Al-MCM-41 catalyst with various copper species, prepared by a combination of three methods—substitution, ion-exchange, and impregnation, was studied for the selective catalytic reduction (SCR) of NO with NH3 at 300 °C for 150 min. In order to investigate the effects of Zn introduction on the nature of the catalyst, Cu/core-shell Al-MCM-41 and Zn/core-shell Al-MCM-41 catalysts were also studied. The roles of Zn promoter in the acidity and the NOx adsorption properties of the catalysts were investigated by in situ Fourier transform infrared spectroscopy (FTIR) of NH3 and NOx adsorption, and temperature-programmed desorption (TPD) of NH3 and NOx. The results demonstrated that the acidity of the catalyst was enhanced by the Zn introduction, as exchanged Zn(II) cations loosely bonded with Al-O-Si framework could create Brønsted acid sites by interacting with OH groups. Moreover, Zn species also provided the additional sites for NO adsorption in the form of nitrite (NO2–) and nitrate (NO3–) species, which are the key intermediates for SCR reaction. In addition, the effect of Zn on the nature of copper was studied by in situ FTIR of CO adsorption and in situ X-ray adsorption near edge structure (XANES). It was found that Zn species hindered the reduction of Cu(II) to Cu(0), resulting in higher Cu(I) species in the Zn promoted catalyst. The Cu-Zn/core-shell Al-MCM-41 exhibited higher catalytic activity compared with that of the Cu/core-shell Al-MCM-41 for the whole reaction time, as it possesses the highest amount of Cu(I) sites, which are responsible for SCR catalytic activity. The Cu-Zn/core-shell Al-MCM-41 catalyst also reached the maximum NO conversion of 100% with the average NO conversion of 76 %. The catalytic performance of the catalyst was further improved by using Zn promoter in the form of ZnO instead of reduced Zn species. The Cu-ZnO/core-shell Al-MCM-41 catalyst showed better catalytic performance with longer working reaction time, and achieved the average NO conversion of 81%.

Keywords: Al-MCM-41, copper, nitrogen oxide, selective catalytic reduction, zinc

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Authors: Elif Coskun Daggecen, Seyma Dokucu, Yekta Gezginc, Ismail Akyol

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Fermented dairy food quality is mainly determined by the sensory perception and influenced by many factors. Today, starter cultures for fermented foods are being developed to have a constant quality in these foods. Streptococcus thermophilus is one of the main species of most a starter cultures of yogurt fermentation. This species produces lactate by lactose fermentation from pyruvate. On the other hand, a small amount of pyruvate can alternatively be converted to various typical yoghurt flavor compounds such as diacetyl, acetoin, acetaldehyde, or acetic acid, for which the activity of three genes are shown to be especially important; ldh, nox and als. Up to date, commercially produced yoghurts have not yet met the desired aromatic properties that Turkish consumers find in traditional homemade yoghurts. Therefore, it is important to select starters carrying favorable metabolic characteristics from natural isolates. In this study, 30 strains of Str. Thermophilus were isolated from traditional Turkish yoghurts obtained from different regions of the country. In these strains, transcriptional levels of ldh, nox and als genes were determined via a newly developed qPCR protocol, which is a more reliable and precision method for analyzing the quantitative and qualitative expression of specific genes in different experimental conditions or in different organisms compared to conventional analytical methods. Additionally, the metabolite production potentials of the isolates were measured. Of all the strains examined, 60% were found to carry the metabolite production potential and the gene activity which appeared to be suitable to be used as a starter culture. Probable starter cultures were determined according to real-time PCR results.

Keywords: gene expression, RT-PCR, starter culture, Streptococcus thermophilus

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Authors: Aydan Atalar, Nurcan Cetinkaya

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Corn stover is leftover of the leaves, stalk, husks and tassels in the field after harvesting the grain combined. Corn stover is a low-quality roughage but has mostly been used as roughage source for feeding ruminant animals in developing countries including Turkey; however, it can also be used to make biofuels as in developed countries. The objectives of the present study were to improve the digestibility of corn stover by the treatment of white rod fungus mainly Pleurotus osteritus (PO), Pleurotus eryingii (PE) and Lantinula edudes (LE) at different incubation times and also to determine the most effective fungus and incubation time to prepare fermeted corn stover for ruminant nutrition. The choped corn stover was treated with PO, PE and LE and incubated for 10, 20, 30 and 40 days in incubator at 26 0C. After each incubation time dry matter(DM), organic matter(OM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), neutral detergent lignin (ADL), in-vitro true dry matter digestibility (IVTDMD) and organic matter digestibility (IVTOMD) were determined. The mean IVTDMD and IVTOMD levels were increased by PO, PE and LE treatments in increasing order of incubation times. The obtained IVTDM values were 59.45, 60.51, 60.82 and 60.18 %; 59.45, 70.55, 67.18 and 66.96 %; 59.45, 70.55, 67.18 and 66,96 %; 59.45, 74.90, 69.18 % ; 59.45, 76.50, 71.24 and 73.04 for control, PO, PE and LE treatments at 0, 10, 20, 30 and 40 days incubation times respectively. The obtained IVTOMD values were 56.45,60.26,60.82and 60.18 %; 56.45, 68.70, 67.18 and 66.96 %; 56.45, 71.26, 69.18 and 69.28 %; 56.45, 73.23, 71.24 and 73.04 % for control, PO, PE and LE treatments at 0, 10, 20, 30 and 40 days incubation times respectively. The most effective fungus was PO and the incubation time was 30 days. In conclusion, PO treatment of corn stover with 30 days incubation may be used to prepare fermented corn stover for ruminant nutrition.

Keywords: biological treatment, corn stover, digestibility, Lantinula edudes, Pleurotus eryingii, Pleurotus osteritus

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Authors: Radhika Jain, Sangeeta Goomer

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In the present study an attempt has been made to re-engineer traditional wadi into wholesome ready-to-use cereal-pulse-based chunks rich in protein quality and fibre content. Chunks were made using extrusion-dehydration combination. Two formulations i.e., whole green gram dhal with instant oats and washed green gram dhal with whole oats were formulated. These chunks are versatile in nature as they can be easily incorporated in day-to-day home-made preparations such as pulao, potato curry and kadhi. Cereal-pulse ratio was calculated using NDpCal%. Limiting amino acids such as lysine, tryptophan, methionine, cysteine and threonine were calculated for maximum amino acid profile in cereal-pulse combination. Time-temperature combination for extrusion at 130^oC and dehydration at 65^oC for 7 hours and 15 minutes were standardized to obtain maximum protein and fibre content. Proximate analysis such as moisture, fat and ash content were analyzed. Protein content of formulation was 62.10% and 68.50% respectively. Fibre content of formulations was 2.99% and 2.45%, respectively. Using a 5-point hedonic scale, consumer preference trials of 102 consumers were conducted and analyzed. Evaluation of chunks prepared in potato curry, kadi and pulao showed preferences for colour 82%, 87%, 86%, texture and consistency 80%, 81%, 88%, flavour and aroma 74%, 82%, 86%, after taste 70%, 75%, 86% and overall acceptability 77%, 75%, 88% respectively. High temperature inactivates antinutritional compounds such as trypsin inhibitors, lectins, saponins etc. Hence, availability of protein content was increased. Developed products were palatable and easy to prepare.

Keywords: extrusion, NDpCal%, protein quality, wadi

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Authors: Pradeep Lamichhane

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Plasma-assisted nitrogen fixation is a process by which atomic nitrogen generated by plasma is converted into ammonia (NH₃) or related nitrogenous compounds. Nitrogen fixation is essential to plant because fixed inorganic nitrogen compounds are required to them for the biosynthesis of all nitrogen-containing organic compounds, such as amino acids and proteins, nucleoside triphosphates and nucleic acid. Most of our atmosphere is composed of nitrogen; however, the plant cannot absorb it directly from the air ambient. As a portion of the nitrogen cycle, nitrogen fixation fundamental for agriculture and the manufacture of fertilizer. In this study, plasma-assisted nitrogen fixation was performed by exposing a non-thermal atmospheric pressure nitrogen plasma generated a sinusoidal power supply (with an applied voltage of 10 kV and frequency of 33 kHz) on a water surface. Besides this, UV excitation of water molecules at the water interface was also done in order to disassociate water. Hydrogen and hydroxyl radical obtained from this UV photolysis electrochemically combine with nitrogen atom obtained from plasma. As a result of this, nitrogen fixation on plasma-activated water (PAW) significantly enhanced. The amount of nitrogen-based products like NOₓ and ammonia (NH₃) synthesized by this combined process of UV and plasma are 1.4 and 2.8 times higher than those obtained by plasma alone. In every 48 hours, 20 ml of plasma-activated water (pH≈3.15) for 10 minutes with moderate concentrations of NOₓ, NH₃ and hydrogen peroxide (H₂O₂) was irrigated on each corn plant (Zea Mays). It was found that the PAW has shown a significant impact on seeds germination rate and improved seedling growth. The result obtained from this experiment suggested that crop yield could increase in a short duration. In the future, this experiment could open boundless opportunities in plasma agriculture to mobilize nitrogen because nitrite, nitrate, and ammonia are more suitable for plant uptake.

Keywords: plasma-assisted nitrogen fixation, nitrogen plasma, UV excitation of water, ammonia synthesis

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Authors: Abduraheem K., Suboohi Nasrin

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Herbaria are specimens of preserved plants, which are very delicate and cellulosic in nature. While these collections are very useful for the enrichment of knowledge and are considered as natural heritage of our entire world, it is very important to preserve and conserve them. The significance is not only to prevent the herbaria from the deterioration of biological agencies but also to preserve its colours and retain natural colour. Colour is not only characteristic of a plant, but it can also help to identify closely related species or to distinguish a plant from a collection of herbaria. Keeping this in mind, a selective solution has been prepared for the conservation and preservation of herbarium in the present study. In this, the quantity of all the selected chemicals, i.e., formaldehyde and copper sulphate was kept constant, and the solution was prepared by dissolving it in distilled water by increasing the amount of picric acid (1, 2, 3, 4, and 5 ml). Fresh specimens of roses and bougainvillea were washed with distilled water and kept in the above solution for 10 to 15 minutes at room temperature. After 10 minutes, the specimen was removed from the solution, dried with the help of paper, and then pressed under the plant press. Blotting sheets were used to absorb the moisture content and were changed every 2 to 3 days to protect against fungal growth. The results revealed that all solutions had insecticidal properties and protected the herbarium specimen against pests. While in the case of colour retention, solution-1 and 2 were not satisfactory colour preservation, and solutions-3 and 5 maintained the colour of rose and bougainvillea leaves for 15 to 20 days and for a month, respectively. After that, the colour begins to fade, and the process is faster in rose leaves than in bougainvillea. And it was also observed that the colour of young leaves started to fade before that of older leaves. When the leaves of rose and bougainvillea are treated with Solution-4, then the colour of rose leaves is maintained for six months.

Keywords: solutions, colour retention, preservation and conservation, leaves of roses and bougainvillea

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Authors: Marwa A. A. Ragab, Amira F. El-Yazbi, Amr El-Hawiet

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The analysis of heavy metals in honey samples is crucial. When found in honey, they denote environmental pollution. Some of these heavy metals as lead either present at low or high concentrations are considered to be toxic. Other heavy metals, for example, copper and zinc, if present at low concentrations, they considered safe even vital minerals. On the contrary, if they present at high concentrations, they are toxic. Their voltammetric determination in honey represents a challenge due to the presence of other electro-active components as vitamins, which may overlap with the peaks of the metal, hindering their accurate and precise determination. The simultaneous analysis of some vitamins: nicotinic acid (B3) and riboflavin (B2), and heavy metals: lead, cadmium, and zinc, in honey samples, was addressed. The analysis was done in 0.1 M Potassium Chloride (KCl) using a hanging mercury drop electrode (HMDE), followed by chemometric manipulation of the voltammetric data using the derivative method. Then the derivative data were convoluted using discrete Fourier functions. The proposed method allowed the simultaneous analysis of vitamins and metals though their varied responses and sensitivities. Although their peaks were overlapped, the proposed chemometric method allowed their accurate and precise analysis. After the chemometric treatment of the data, metals were successfully quantified at low levels in the presence of vitamins (1: 2000). The heavy metals limit of detection (LOD) values after the chemometric treatment of data decreased by more than 60% than those obtained from the direct voltammetric method. The method applicability was tested by analyzing the selected metals and vitamins in real honey samples obtained from different botanical origins.

Keywords: chemometrics, overlapped voltammetric peaks, derivative and convoluted derivative methods, metals and vitamins

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Authors: Kubra Dogan, Fatih Tornuk

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Due to increasing consumer demand for the high-quality food products and awareness regarding the health benefits of different nutrients in food minimal processing becomes more popular in modern food preservation. To date, heat treatment is often used for inactivation of spoilage microorganisms in foods. However, it may cause significant changes in the quality and nutritional properties of food. In order to overcome the detrimental effects of heat treatment, several alternatives of non-thermal microbial inactivation processes have been investigated. Ultraviolet (UV) inactivation is a promising and feasible method for better quality and longer shelf life as an alternative to heat treatment, which aims to inhibit spoilage and pathogenic microorganisms and to inactivate the enzymes in vegetable juice production. UV-C is a sub-class of UV treatment which shows the highest microcidal effect between 250-270 nm. The wavelength of 254 nm is used for the surface disinfection of certain liquid food products such as vegetable juice. Effects of UV-C treatment on microbiological load and quality parameter of vegetable juice which is a mix of celery, carrot, lemon and orange was investigated. Our results showed that storing of UV-C applied vegetable juice for three months, reduced the count of TMAB by 3.5 log cfu/g and yeast-mold by 2 log cfu/g compared to control sample. Total phenolic content was found to be 514.3 ± 0.6 mg gallic acid equivalent/L, and there wasn’t a significant difference compared to control. The present work suggests that UV-C treatment is an alternative method for disinfection of vegetable juice since it enables adequate microbial inactivation, longer shelf life and has minimal effect on degradation of quality parameters of vegetable juice.

Keywords: heat treatment, phenolic content, shelf life, ultraviolet (UV-C), vegetable juice

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Authors: Benyapa Suksuwan

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Introduction: Oxidative stress is identified as the root cause of the development and progression of several diseases as the disproportion of free radicals in the body leads to tissue or cell damage. Polyphenols are the most common antioxidant found in plants and are efficient in capturing oxidative free radicals. Aim of the Study: This study focused on the antioxidant activity of polyphenols extracted from Phyllanthus Emblica L. as oxidative stress plays a vital role in developing and progressing many diseases, including cardiovascular diseases and cancer. Materials and Methods: The plant was extracted using a mixture solvent (ethyl alcohol: water in ratio 8:2). The total phenolic content of P. Emblica extract was determined using the Folin-Cioucalteu method and calculated as gallic acid equivalents (GAE) and various antioxidant assays DPPH and ABTS radical scavenging capacity assays. Results and Discussion: The findings exhibited a strong correlation between antioxidant activity and the total phenol contents. In addition, the IC₅₀ of P. Emblica extract via DPPH and ABTS assays were 68.10 μg/mL ± 0.455, and 49.24 μg/mL ± 0.716, respectively. Furthermore, P. Emblica extract showed antioxidant activities in a concentration-dependent manner. Vitamin C was used as a positive control in the DPPH assay, while Trolox was used as a positive control in the ABTS assay. Conclusions: In conclusion, P. Emblica extract consisted of a high amount of total phenolic content, which possesses potent antioxidant activity. However, further antioxidant activity assays using human cell lines such as SOD, ROS, and RNS scavenging assays and in vitro antioxidant experiments should be performed in order.

Keywords: antioxidant, ABTS scavenging, DPPH scavenging assay, total phenol contents assay, Phyllanthus Emblica L

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Authors: F. Al-Sheikh, C. Moralejo, M. Pritzker, W. A. Anderson, A. Elkamel

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Ammonia in mining wastewater is a significant problem, and treatment can be especially difficult in cold climates where biological treatment is not feasible. An adsorption process is one of the alternative processes that can be used to reduce ammonia concentrations to acceptable limits, and therefore a LEWATIT resin strongly acidic H+ form ion exchange resin and a Bowie Chabazite Na form AZLB-Na zeolite were tested to assess their effectiveness. For these adsorption tests, two packed bed columns (a mini-column constructed from a 32-cm long x 1-cm diameter piece of glass tubing, and a 60-cm long x 2.5-cm diameter Ace Glass chromatography column) were used containing varying quantities of the adsorbents. A mining wastewater with ammonia concentrations of 22.7 mg/L was fed through the columns at controlled flowrates. In the experimental work, maximum capacities of the LEWATIT ion exchange resin were 0.438, 0.448, and 1.472 mg/g for 3, 6, and 9 g respectively in a mini column and 1.739 mg/g for 141.5 g in a larger Ace column while the capacities for the AZLB-Na zeolite were 0.424, and 0.784 mg/g for 3, and 6 g respectively in the mini column and 1.1636 mg/g for 38.5 g in the Ace column. In the theoretical work, Thomas, Adams-Bohart, and Yoon-Nelson models were constructed to describe a breakthrough curve of the adsorption process and find the constants of the above-mentioned models. In the regeneration tests, 5% hydrochloric acid, HCl (v/v) and 10% sodium hydroxide, NaOH (w/v) were used to regenerate the LEWATIT resin and AZLB-Na zeolite with 44 and 63.8% recovery, respectively. In conclusion, continuous flow adsorption using a LEWATIT ion exchange resin and an AZLB-Na zeolite is efficient when using a co-flow technique for removal of the ammonia from wastewater. Thomas, Adams-Bohart, and Yoon-Nelson models satisfactorily fit the data with R² closer to 1 in all cases.

Keywords: AZLB-Na zeolite, continuous adsorption, Lewatit resin, models, regeneration

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Authors: Gloria Pinilla, Jose Manuel Baena, Patricia Gálvez-Martín, Juan Antonio Marchad

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Cartilage is a dense connective tissue with limited self-repair properties. Currently, the therapeutic use of autologous or allogenic chondrocytes makes up an alternative therapy to the pharmacological treatment. The design of a bioprinted 3D cartilage with chondrocytes and biodegradable biomaterials offers a new therapeutic alternative able of bridging the limitations of current therapies in the field. We have developed an enhanced printing processes-Injection Volume Filling (IVF) to increase the viability and survival of the cells when working with high-temperature thermoplastics without the limitation of the scaffold geometry in contact with cells. We have demonstrated the viability of the printing process using chondrocytes for cartilage regeneration. This development will accelerate the clinical uptake of the technology and overcomes the current limitation when using thermoplastics as scaffolds. An alginate-based hydrogel combined with human chondrocytes (isolated from osteoarthritis patients) was formulated as bioink-A and the polylactic acid as bioink-B. The bioprinting process was carried out with the REGEMAT V1 bioprinter (Regemat 3D, Granada-Spain) through a IVF. The printing capacity of the bioprinting plus the viability and cell proliferation of bioprinted chondrociytes was evaluated after five weeks by confocal microscopy and Alamar Blue Assay (Biorad). Results showed that the IVF process does not decrease the cell viability of the chondrocytes during the printing process as the cells do not have contact with the thermoplastic at elevated temperatures. The viability and cellular proliferation of the bioprinted artificial 3D cartilage increased after 5 weeks. In conclusion, this study demonstrates the potential use of Regemat V1 for 3D bioprinting of cartilage and the viability of bioprinted chondrocytes in the scaffolds for application in regenerative medicine.

Keywords: cartilage regeneration, bioprinting, bioink, scaffold, chondrocyte

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Authors: Przemyslaw Nogly, Tobias Weinert, Daniel James, Sergio Carbajo, Dmitry Ozerov, Antonia Furrer, Dardan Gashi, Veniamin Borin, Petr Skopintsev, Kathrin Jaeger, Karol Nass, Petra Bath, Robert Bosman, Jason Koglin, Matthew Seaberg, Thomas Lane, Demet Kekilli, Steffen Brünle, Tomoyuki Tanaka, Wenting Wu, Christopher Milne, Thomas A. White, Anton Barty, Uwe Weierstall, Valerie Panneels, Eriko Nango, So Iwata, Mark Hunter, Igor Schapiro, Gebhard Schertler, Richard Neutze, Jörg Standfuss

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Ultrafast isomerization of retinal is the primary step in a range of photoresponsive biological functions including vision in humans and ion-transport across bacterial membranes. We studied the sub-picosecond structural dynamics of retinal isomerization in the light-driven proton pump bacteriorhodopsin using an X-ray laser. Twenty snapshots with near-atomic spatial and temporal resolution in the femtosecond regime show how the excited all-trans retinal samples conformational states within the protein binding pocket prior to passing through a highly-twisted geometry and emerging in the 13-cis conformation. The aspartic acid residues and functional water molecules in proximity of the retinal Schiff base respond collectively to formation and decay of the initial excited state and retinal isomerization. These observations reveal how the protein scaffold guides this remarkably efficient photochemical reaction.

Keywords: bacteriorhodopsin, free-electron laser, retinal isomerization mechanism, time-resolved crystallography

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Authors: Katarzyna Miśkiewicz, Magdalena Lasoń-Rydel, Małgorzata Krępska, Katarzyna Sieczyńska, Iwona Masłowska-Lipowicz, Katarzyna Ławińska

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The aim of the study was to analyse a selection of 12 pasteurised products made from fruit and vegetables, such as fruit juices, fruit drinks, jams, marmalades and jam produced by small regional factories as well as large food corporations. The research was carried out as part of the project "Innovative system of healthy and regional food distribution", funded by the Ministry of Education and Science (Poland), which aims to create an economically and organisationally strong agri-food industry in Poland through effective cooperation between scientific and socio-economic actors. The main activities of the project include support for the creation of new distribution channels for regional food products and their easy access to a wide group of potential customers while maintaining the highest quality standards. One of the key areas of the project is food quality analyses conducted to indicate the competitive advantage of regional products. Presented here are studies on the content of sugars, vitamin C, preservatives, synthetic colours, sweeteners, sodium and potassium, as well as studies on the microbiological purity of selected products made from fruit and vegetables. The composition of products made from fruit and vegetables varies greatly and depends on both the type of raw material and the way it is processed. Of the samples tested, fruit drinks contained the least amount of sugars, and jam and marmalade made by large producers and bought in large chain stores contained the most. However, the low sugar content of some fruit drinks is due to the presence of the sweetener sucralose in their composition. The vitamin C content of the samples varied, being higher in products where it was added during production. All products made in small local factories were free of food additives such as preservatives, sweeteners and synthetic colours, indicating their superiority over products made by large producers. Products made in small local factories were characterised by a relatively high potassium content. The microbiological purity of commercial products was confirmed - no Salmonella spp. were detected, and the number of mesophilic bacteria, moulds, yeasts, and β-glucuronidase-positive E. coli was below the limit of quantification.

Keywords: fruit and vegetable products, sugars, food additives, HPLC, ICP-OES

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Authors: Sara Przetocka, Krzysztof Żak, Grzegorz Dubin, Tadeusz Holak

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Toll-like receptors (TLRs) play central role in the innate immune response and inflammation by recognizing pathogen-associated molecular patterns (PAMPs). A fundamental basis of TLR signalling is dependent upon the recruitment and association of adaptor molecules that contain the structurally conserved Toll/interleukin-1 receptor (TIR) domain. MyD88 (myeloid differentiation primary response gene 88) is the universal adaptor for TLRs and cooperates with Mal (MyD88 adapter-like protein, also known as TIRAP) in TLR4 response which is predominantly used in inflammation, host defence and carcinogenesis. Up to date two possible models of MyD88, Mal and TLR4 interactions have been proposed. The aim of our studies is to confirm or abolish presented models and accomplish the full structural characterisation of TIR domains interaction. Using molecular cloning methods we obtained several construct of MyD88 and Mal TIR domain with GST or 6xHis tag. Gel filtration method as well as pull-down analysis confirmed that recombinant TIR domains from MyD88 and Mal are binding in complexes. To examine whether obtained complexes are homo- or heterodimers we carried out cross-linking reaction of TIR domains with BS3 compound combined with mass spectrometry. To investigate which amino acid residues are involved in this interaction the NMR titration experiments were performed. 15N MyD88-TIR solution was complemented with non-labelled Mal-TIR. The results undoubtedly indicate that MyD88-TIR interact with Mal-TIR. Moreover 2D spectra demonstrated that simultaneously Mal-TIR self-dimerization occurs which is necessary to create proper scaffold for Mal-TIR and MyD88-TIR interaction. Final step of this study will be crystallization of MyD88 and Mal TIR domains complex. This crystal structure and characterisation of its interface will have an impact in understanding the TLR signalling pathway and possibly will be used in development of new anti-cancer treatment.

Keywords: cancer, MyD88, TIR domains, Toll-like receptors

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Authors: Niang Mamadou Sileye

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The study turns on the use of the phytase in aquaculture, its zootechnical interests and the possibilities of incorporation in the feed. The goal is to reduce the waste in phosphorus linked to the feeding of fishes, without any loss of zootechnical performances and with a decrease of feed costs. We have studied the literature in order to evaluate the raw materials (total phosphorus, phytate and available phosphorus) used by a company to manufacture feed for rainbow trout; to determine the phosphorus requirements for aquaculture species; to determine the requirements of phosphorus for aquaculture species, to determine the sings of lack of phosphorus for fishes; to study the antagonism between the phosphorus and the calcium and to study also the different forms of waste for the rainbow trout. The results found in the bibliography enable us test several Hypothesis of feed formulation for rainbow trout with different raw materials. This simulation and the calculation for wastes allowed to validate two formulation of feed: a control feed (0.5% of monocalcique phosphate) and a trial feed (supplementation with 0.002% of phytase Ronozyme PL and without inorganic phosphate). The feeds have been produced and sent to a experimental structure (agricultural college of Brehoulou).The result of the formulation give a decrease of the phosphorus waste of 28% for the trial feed compared to the feed. The supplementation enables a gain of 2.3 euro per ton. The partial results of the current test show no significant difference yet for the zootechnical parameters (growth rate, mortality, weight gain and obvious conversion rate) between control feed and the trial one. The waste measures do not show either significant difference between the control feed and the trial one, but however, the average difference would to decrease the wastes of 35.6% thanks to the use of phytase.

Keywords: phosphorus, phytic acid, phytase, need, digestibility, formulation, food, waste, rainbow trout

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Authors: E. Chávez-Vargas, M. de la L. Olvera-Amador, A. Jimenez-Gonzalez, A. Maldonado

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Undoped and fluorine doped zinc oxide (ZnO) thin films were deposited on sodocalcic glass substrates by the ultrasonic chemical spray technique. As the main goal is the manufacturing of transparent electrodes, the effects of both the solution composition and the substrate temperature on both the electrical and optical properties of ZnO thin films were studied. As a matter of fact, the effect of fluorine concentration ([F]/[F+Zn] at. %), solvent composition (acetic acid, water, methanol ratios) and ageing time, regarding solution composition, were varied. In addition, the substrate temperature and the deposition time, regarding the chemical spray technique, were also varied. Structural studies confirm the deposition of polycrystalline, hexagonal, wurtzite type, ZnO. The results show that the increase of ([F]/[F+Zn] at. %) ratio in the solution, decreases the sheet resistance, RS, of the ZnO:F films, reaching a minimum, in the order of 1.6 Ωcm, at 60 at. %; further increase in the ([F]/[F+Zn]) ratio increases the RS of the films. The same trend occurs with the variation in substrate temperature, as a minimum RS of ZnO:F thin films was encountered when deposited at TS= 450 °C. ZnO:F thin films deposited with aged solution show a significant decrease in the RS in the order of 100 ΩS. The transmittance of the films was also favorable affected by the solvent ratio and, more significantly, by the ageing of the solution. The whole evaluation of optical and electrical characteristics of the ZnO:F thin films deposited under different conditions, was done under Haacke’s figure of Merit in order to have a clear and quantitative trend as transparent conductors application.

Keywords: zinc oxide, ZnO:F, TCO, Haacke’s figure of Merit

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Authors: Ume Habiba, Shafaqat Ali, Mujahid Farid, Muhammad Bilal Shakoor

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Copper (Cu) is an essential micronutrient for normal plant growth and development, but in excess, it is also toxic to plants. The present study investigated the influence of ethylenediaminetetraacetic acid (EDTA) in enhancing Cu uptake and tolerance as well as the morphological and physiological responses of Brassica napus L. seedlings under Cu stress. Four-week-old seedlings were transferred to hydroponics containing Hoagland’s nutrient solution. After 2 weeks of transplanting, three levels (0, 50, and 100 μM) of Cu were applied with or without application of 2.5 mM EDTA and plants were further grown for 8 weeks in culture media. Results showed that Cu alone significantly decreased plant growth, biomass, photosynthetic pigments, and gas exchange characteristics. Cu stress also reduced the activities of antioxidants, such as superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT) along with protein contents. Cu toxicity increased the concentration of reactive oxygen species (ROS) as indicated by the increased production of malondialdehyde (MDA) and hydrogen peroxide (H2O2) in both leaves and roots. The application of EDTA significantly alleviated Cu-induced toxic effects in B. napus, showing remarkable improvement in all these parameters. EDTA amendment increased the activity of antioxidant enzymes by decreasing the concentrations of MDA and H2O2 both in leaves and roots of B. napus. Although, EDTA amendment with Cu significantly increased Cu uptake in roots, stems, and leaves in decreasing order of concentration but increased the growth, photosynthetic parameters, and antioxidant enzymes. These results showed that the application of EDTA can be a useful strategy for phytoextraction of Cu by B. napus from contaminated soils.

Keywords: antioxidants, biomass, copper, EDTA, phytoextraction, tolerance

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Authors: Thuy Phuong Nhat Tran, Ashutosh Thakur, Toshiaki Taniike

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Recently, covalently linked graphene oxide frameworks (GOFs) have attracted considerable attention in gas absorbance and water purification as well-defined microporous materials. In spite of their potential advantages such as a controllable pore dimension, adjustable hydrophobicity, and structural stability, these materials have been scarcely employed in heterogeneous catalysis. Here we demonstrate a novel and facile method to synthesize Pd nanoparticles (NPs) confined in a GOF (Pd@GOF). The GOF with uniform interlayer space was obtained by the intercalation of diboronic acid between graphene oxide layers. It was found that Pd NPs were generated inside the graphitic gallery spaces of the GOF, and thus, formed Pd NPs were well-dispersed with a narrow particle size distribution. The synthesized Pd@GOF emerged as an efficient nanocatalyst based on its superior performance (product yield and recyclability) toward Suzuki-Miyaura cross-coupling reaction in both polar and apolar solvents, which has been hardly observed for previously reported graphene-based Pd nanocatalysts. Furthermore, the rational comparison of the catalytic performance between two kinds of Pd@GOF (Pd NPs encapsulated in a diboronic ester-intercalated GOF and in a monoboronic ester-intercalated GOF) firmly confirmed the essential role of a rigid framework design in the stabilization of Pd NPs. Based on these results, the covalently assembled GOF was proposed as a promising scaffold for hosting noble metal NPs to construct desired metal@GOF nanocatalysts with improved activity and durability.

Keywords: graphene oxide framework, palladium nanocatalyst, pore confinement, Suzuki-Miyaura cross-coupling reaction

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Authors: Wittaya Apai, Satippong Rattanakam, Suttinee Likhittragulrung, Nuttanai Tungmunkongvorakul, Sompetch Jaroensuk

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The objective of this study was to find some alternative ways to decrease sulfur dioxide (SO₂) residue problem and prolong storage life in fresh longan for export. Office of Agricultural Research and Development Region 1, Chiang Mai province conducted the research and development from 2016-2018. A grade longan cv. Daw fruit with panicle attached was placed in 11.5 kg commercial perforated plastic basket. They had 5 selected treatments comprising of 3 baskets as replication for each treatment, i.e. 1.5% SO₂ fumigation prior to insert SO₂-generated pads (Uvasys®) (1.5% SO₂+SO₂ pad), dipping in 5% hydrochloric acid (HCl) mixed with 1% sodium metabisulfite (SMS) for 5 min (5% HCl +1% SMS), ozone (O₃) fumigation for 1 hours (h) prior to 1.5% SO₂ fumigation (O₃ 1 h+1.5% SO₂), 1.5% SO₂ fumigation prior to O₃ fumigation for 1 h (1.5% SO₂+O₃ 1 h) and 1.5% SO₂ fumigation alone as commercial treatment (1.5% SO₂). They were stored at 5 ˚C, 90% relative humidity (RH) for 40-80 days. The results found that the possible treatments were 1.5% SO₂+O₃ 1 h and 5% HCl +1% SMS respectively and prevented pericarp browning for 80 days at 5 ºC. There were no significant changes in some parameters in any treatments; 1.5% SO₂+O₃ 1 h and 1.5% SO₂ during storage, i.e., pericarp browning, flesh discoloration, disease incidence (%) and sensory evaluation during storage. Application 1.5% SO₂+O₃ 1 h had a tendency less both SO₂ residue in fruit and disease incidence (%) including brighter pericarp color as compared with commercial 1.5% SO₂ alone. Moreover, HCl 5%+SMS 1% showed the least SO₂ residue in whole fruit below codex tolerance at 50 mg/kg throughout period of time. The fruit treated with 1.5% SO₂+O₃ 1 h, 1.5% SO₂, 5% HCl+1% SMS, O₃ 1 h+1.5% SO₂, and 1.5% SO₂+SO₂ pad could prolong storage life for 40, 40, 40, 30 and 30 days respectively at 5°C, 90% RH. Thus, application 1.5% SO₂+O₃ 1 h and/or 5% HCl +1% SMS could be used for extending shelf life fresh longan exported to restricted countries due to less SO₂ residue and fruit quality was maintained as compared with the conventional method.

Keywords: longan, sulfur dioxide, ozone fumigation, sodium metabisulfite

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Authors: Omoruyi Gold Idemudia, Alexander P. Sadimenko

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The synthesis of N-heterocycles with novel properties, having broad spectrum biological activities that may become alternative medicinal drugs, have been attracting a lot of research attention due to the emergence of medicinal drug’s limitations such as disease resistance and their toxicity effects among others. Acylpyrazolones have been employed as pharmaceuticals as well as analytical reagent and their application as coordination complexes with transition metal ions have been well established. By way of a condensation reaction with amines acylpyrazolone ketones form a more chelating and superior group of compounds known as azomethines. 4-propyl-3-methyl-1-phenyl-2-pyrazolin-5-one was reacted with phenylhydrazine to get a new phenylhydrazone which was further reacted with aqueous solutions of palladium and platinum salts, in an effort towards the discovery of transition metal based synthetic drugs. The compounds were characterized by means of analytical, spectroscopic, thermogravimetric analysis TGA, as well as x-ray crystallography. 4-propyl-3-methyl-1-phenyl-2-pyrazolin-5-one phenylhydrazone crystallizes in a triclinic crystal system with a P-1 (No. 2) space group based on x-ray crystallography. The bidentate ON ligand formed a square planar geometry on coordinating with metal ions based on FTIR, electronic and NMR spectra as well as magnetic moments. Reported compounds showed antibacterial activities against the nominated bacterial isolates using the disc diffusion technique at 20 mg/ml in triplicates. The metal complexes exhibited a better antibacterial activity with platinum complex having an MIC value of 0.63 mg/ml. Similarly, ligand and complexes also showed antioxidant scavenging properties against 2, 2-diphenyl-1-picrylhydrazyl DPPH radical at 0.5mg/ml relative to ascorbic acid (standard drug).

Keywords: acylpyrazolone, antibacterial studies, metal complexes, phenylhydrazone, spectroscopy

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Authors: Norah Al Hokbany, Ibrahim Al Jammaz, Basem Al Otaibi, Yousif Al Malki, Subhani M. Okarvi

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Objective: The folate receptor is over-expressed in a wide variety of human tumors. Conjugates of folate have been shown to be selectively taken up by tumor cells via the folate receptor. In an attempt to develop new folate radiotracers with favorable biochemical properties for detecting folate receptor-positive cancers. Methods: we synthesized ⁶⁴Cu-NOTA- and ⁶⁴Cu-NOTAM-folate conjugates using a straightforward and simple one-step reaction. Radiochemical yields were greater than 95% (decay-corrected) with a total synthesis time of less than 20 min. Results: Radiochemical purities were always greater than 98% without high-performance liquid chromatography (HPLC) purification. These synthetic approaches hold considerable promise as a rapid and simple method for ⁶⁴Cu-folate conjugate preparation with high radiochemical yield in a short synthesis time. In vitro tests on the KB cell line showed that significant amounts of the radio conjugates were associated with cell fractions. Bio-distribution studies in nude mice bearing human KB xenografts demonstrated a significant tumor uptake and favorable bio-distribution profile for ⁶⁴Cu-NOTA- and ⁶⁴Cu-NOTAM-folate conjugate. The uptake in the tumors was blocked by the excess injection of folic acid, suggesting a receptor-mediated process. Conclusion: These results demonstrate that the ⁶⁴Cu-NOTAM-folate conjugate may be useful as a molecular probe for the detection and staging of folate receptor-positive cancers, such as ovarian cancer and their metastasis, as well as monitoring tumor response to treatment.

Keywords: folate, receptor, tumor imaging, ⁶⁴Cu-NOTA-folate, PET

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Authors: Muhammad Sajid Arshad, Sadaf Bashir

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In this study, the effects of pomegranate peel based edible coating were determined on safety and quality of chicken nuggets. Four treatment groups were prepared as control (without coating), coating with sodium alginate (SA) (1.5%), pomegranate peel powder (PPP) (1.5%), and combination of SA and PPP. There was a significant variation observed with respect to coating treatments and storage intervals. The chicken nuggets were subjected to refrigerated storage (40C) and were analyzed at regular intervals of 0, 7, 14 1 and 21 days. The microbiological quality was determined by total aerobic and coliform counts. Total aerobic (5.09±0.05 log CFU/g) and coliforms (3.91±0.06 log CFU/g) counts were higher in uncoated chicken nuggets whereas lower was observed in coated chicken nuggets having combination of SA and PPP. Likewise, antioxidants potential of chicken nuggets was observed by assessing total phenolic contents (TPC) and DPPH activity. Higher TPC (135.66 GAE/100g) and DPPH (64.65%) were found in combination with SA and PPP, whereas minimum TPC (91.38) and DPPH (41.48) was observed in uncoated chicken nuggets. Regarding the stability analysis of chicken nuggets, thiobarbituric acid reactive substances (TBARS) and peroxide value (POV) were determined. Higher TBARS (1.62±0.03 MDA/Kg) and POV (0.92±0.03 meq peroxide/kg) were found in uncoated chicken nuggets. Hunter color values were also observed in both uncoated and coated chicken nuggets. Sensorial attributes were also observed by the trained panelists. The higher sensory score for appearance, color, taste, texture and overall acceptability were observed in control (uncoated) while in coated treatments, it was found within acceptable limits. In nutshell, the combination of SA and PPP enhanced the overall quality, antioxidant potential, and stability of chicken nuggets.

Keywords: chicken nuggets, edible coatings, pomegranate peel powder, sodium alginate

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Authors: Arun K. Yadav, Adam J. Carroll, Gonzalo M. Estavillo, Greg J. Rebetzke, Barry J. Pogson

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Water limits crop productivity, so selecting for minimal yield-gap in drier environments is critical to mitigate against climate change and land-use pressures. To date, no markers measured in glasshouses have been reported to predict field-based drought tolerance. In the field, the best measure of drought tolerance is yield-gap; but this requires multisite trials that are an order of magnitude more resource intensive and can be impacted by weather variation. We investigated the responses of relative water content (RWC), stomatal conductance (gs), chlorophyll content and metabolites in flag leaves of commercial wheat (Triticum aestivum L.) cultivars to three drought treatments in the glasshouse and field environments. We observed strong genetic associations between glasshouse-based RWC, metabolites and Yield gap-based Drought Tolerance (YDT): the ratio of yield in water-limited versus well-watered conditions across 24 field environments spanning sites and seasons. Critically, RWC response to glasshouse drought was strongly associated with both YDT (r2 = 0.85, p < 8E-6) and RWC under field drought (r2 = 0.77, p < 0.05). Multiple regression analyses revealed that 98% of genetic YDT variance was explained by drought responses of four metabolites: serine, asparagine, methionine and lysine (R2 = 0.98; p < 0.01). Fitted coefficients suggested that, for given levels of serine and asparagine, stronger methionine and lysine accumulation was associated with higher YDT. Collectively, our results demonstrate that high-throughput, targeted metabolic phenotyping of glasshouse-grown plants may be an effective tool for the selection of wheat cultivars with high YDT in the field.

Keywords: drought stress, grain yield, metabolomics, stomatal conductance, wheat

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Authors: M. A. Mamun, M. I. Khan, M. H. Sarker, K. A. Khan, M. Shajahan

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The study was carried out to investigate on an innovative invention, Pathor Kuchi Leaf (PKL) cell, which is fueled with PKL sap of widely available plant called Bryophyllum pinnatum as an energy source for use in PKL battery to generate electricity. This battery, a primary source of electricity, has several order of magnitude longer shelf-lives than the traditional Galvanic cell battery, is still under investigation. In this regard, we have conducted some experiments using various instruments including Atomic Absorption Spectrophotometer (AAS), Ultra-Violet Visible spectrophotometer (UV-Vis), pH meter, Ampere-Volt-Ohm Meter (AVO Meter), etc. The AAS, UV-Vis, and pH-metric analysis data provided that the potential and current were produced as the Zn electrode itself acts as reductant while Cu2+ and H+ ions are behaving as the oxidant. The significant influence of secondary salt on current and potential leads to the dissociation of weak organic acids in PKL juice, and subsequent enrichment to the reactant ions by the secondary salt effects. However, the liquid junction potential was not as great as minimized with the opposite transference of organic acid anions and H+ ions as their dissimilar ionic mobilities. Moreover, the large value of the equilibrium constant (K) implies the big change in Gibbs free energy (∆G), the more electromotive force works in electron transfer during the forward electrochemical reaction which coincides with the fast reduction of the weight of zinc plate, revealed the additional electrical work in the presence of PKL sap. This easily fabricated high-performance PKL battery can show an excellent promise during the off-peak across the countryside.

Keywords: Atomic Absorption Spectrophotometer (AAS), Bryophylum Pinnatum Leaf (BPL), electricity, electrochemistry, organic acids

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Authors: Stafford Stewart, Hui Yu, Rominder Suri

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This paper publishes the results of the first known study conducted in urban Philadelphia waterways that simultaneously utilized anthropogenic chemical and biological markers to identify suitable indicators of untreated human waste and fecal pollution. A total of 13 outfall samples, 30 surface water samples, and 2 groundwater samples were analyzed for fecal contamination and untreated human waste using a suite of 25 chemical markers and 5 bio-markers. Pearson rank correlation tests were conducted to establish associations between the abundances of bio-markers and the concentrations of chemical markers. Results show that 16S rRNA gene of human-associated Bacteroidales (BacH) was very strongly correlated (0.76 – 0.97, p < 0.05) with labile chemical markers acetaminophen, cotinine, estriol, and urobilin. Likewise, human-specific F- RNA coliphages (F-RNA-II) and labile chemical markers, urobilin, ibuprofen, cotinine and estriol, were significantly correlated (0.77 – 0.95, p < 0.05). Similarly, a strong positive correlation (0.67 – 0.91, p < 0.05) was evident between the abundances of bio-markers BacH and F-RNA-II, and the concentrations of the conservative markers, trimethoprim, meprobamate, diltiazem, triclocarban, metformin, sucralose, gemfibrozil, sulfamethoxazole, and carbamazepine. Human mitochondrial DNA (MitoH) correlated moderately with labile markers nicotine and salicylic acid as well as with conservative markers metformin and triclocarban (0.31 – 0.47, p<0.05). This study showed that by associating chemical and biological markers, a robust technique was developed for fingerprinting source-specific untreated waste and fecal contamination in source waters.

Keywords: anthropogenic markers, bacteroidales, fecal pollution, source waters, wastewater

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Authors: Falana Yetunde Olaitan, Ijah U. J. J, Solebo Shakirat O.

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Genetically modified crops are already phenomenally successful and are grown worldwide in more than eighteen countries on more than 67 million hectares. Nigeria, in October 2018, approved Bacillus thuringiensis (Bt) cotton and maize; therefore, the need to carry out environmental risk assessment studies. A total of 15 4L octagonal ceramic pots were filled with 4kg of soil and placed on the bench in 2 rows of 10 pots each and the 3rd row of 5 pots, 1st-row pots were used to plant GM cotton seeds, while the 2nd-row pots were used for non-GM cotton seeds and the 3rd row of 5 pots served as control, all in the screen house. Soil samples for metagenomic DNA extraction were collected at random and at the monthly interval after planting at a distance of 2mm from the plant’s root and at a depth of 10cm using a sterile spatula. Soil samples for physicochemical analysis were collected before planting and after harvesting the GM and non-GM crops as well as from the control soil. The DNA was extracted, quantified and sequenced; Sample 1A (DNA from GM cotton Soil at 1st interval) gave the lowest sequence read with 0.853M while sample 2B (DNA from GM cotton Soil at 2nd interval) gave the highest with 5.785M, others gave between 1.8M and 4.7M. The samples treatment were grouped into four, Group 1 (GM cotton soil from 1 to 3 intervals) had between 800,000 and 5,700,000 strains of microbes (SOM), Group 2 (non GM cotton soil from 1 to 3 intervals) had between 1,400,600 and 4,200,000 SOM, Group 3 (control soil) had between 900,000 and 3,600,000 SOM and Group 4 (initial soil) had between 3,700,000 and 4,000,000 SOM. The microbes observed were predominantly bacteria (including archaea), fungi, dark matter alongside protists and phages. The predominant bacterial groups were the Terrabacteria (Bacillus funiculus, Bacillus sp.), the Proteobacteria (Microvirga massiliensis, sphingomonas sp.) and the Archaea (Nitrososphaera sp.), while the fungi were Aspergillus fischeri and Fusarium falciforme. The comparative analysis between groups was done using JACCARD PERMANOVA beta diversity analysis at P-value not more than 0.76 and there was no significant pair found. The pH for initial, GM cotton, non-GM cotton and control soil were 6.28, 6.26, 7.25, 8.26 and the percentage moisture was 0.63, 0.78, 0.89 and 0.82, respectively, while the percentage Nitrogen was observed to be 17.79, 1.14, 1.10 and 0.56 respectively. Other parameters include, varying concentrations of Potassium (0.46, 1,284.47, 1,785.48, 1,252.83 mg/kg) and Phosphorus (18.76, 17.76, 16.87, 15.23 mg/kg) were recorded for the four treatments respectively. The soil consisted mainly of silt (32.09 to 34.66%) and clay (58.89 to 60.23%), reflecting the soil texture as silty – clay. The results were then tested with ANOVA at less than 0.05 P-value and no pair was found to be significant as well. The results suggest that the GM crops have no significant effect on microbial ecology and physicochemical properties of the soil and, in turn, no direct or indirect effects on human health.

Keywords: genetically modified crop, microbial ecology, physicochemical properties, metagenomics, DNA, soil

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