Search results for: enzymatic pretreatment

Authors: Ouchemoukh Salim, Amessis-Ouchemoukh Nadia, Guenaoui Nawel, Moumeni Lynda, Zaidi Hicham, Otmani Amar, Sadou Dyhia

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Honey is a hive food rich in carbohydrates and water and it also has a lot of nutrients (enzymes, minerals, organic acids, phytochemicals...). It is used in different nutritional and therapeutic fields. Algerian honey was studied for its physicochemical parameters, nutritional values (moisture, brix, pH, electrical conductivity, and amounts of HMF, proteins, proline, total phenolic compounds and flavonoids) and some biological activities (antioxidant, anti-inflammatory and enzymatic anti-browning). The antioxidant activities of the samples were estimated using different methods (ABTS, DPPH free radicals scavenging, reducing power, and chelating ferrous activity). All honeys were acidic (3.45≤pH≤4.65). The color varied from mimosa yellow to dark brown. The specific rotation was levorotatory in most honey samples, and the electrical conductivity, hydroxymethylfurfural, and proline values agreed with the international honey requirements. For anti-inflammatory activity, the results showed that the inhibiting capacity of the denaturation of the BSA of the honey analyzed varied from 15 to 75 % with a maximum of activity at the concentration of 0,5 mg/ml. All honey exhibited enzymatic anti-browning on different slices of fruits. In fact, the results showed that the controls have the greatest browning unit compared to the honeys studied and PPO and POD enzymes had the lowest enzyme activity. High significant correlations were found between the color of honey, its antioxidant content and its biological activities (antioxidant, anti-inflammatory and enzymatic anti-browning). The dark color of honey is a good indicator of the best biological properties, therefore, the best nutritional and therapeutic values.

Keywords: honey, physico-chemical parameters, bioactive compounds, biological properties

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Authors: S. K. Thind, Aparjot Kaur

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Heat stress is one of the major environmental factors drastically reducing wheat production. Crop heat tolerance can be enhanced by preconditioning of plants by exogenous application of osmoprotectants. Presently, the effect of trehalose pretreatment (at 1 mM, and 1.5 nM) under heat stress of 35±2˚C (moderate) and 40±2˚ (severe) for four and eight hour was conducted in wheat (Tricticum aestivum L.) genotypes viz. HD2967, PBW 175, PBW 343, PBW 621, and PBW 590. Heat stress affects wide spectrum of physiological processes within plants that are irreversibly damaged by stress. Membrane thermal stability (MTS) and cell viability was significantly decreased under heat stress for eight hours. Pretreatment with trehalose improved MTS and cell viability under stress and this effect was more promotory with higher concentration. Thermal stability of photosynthetic apparatus differed markedly between genotypes and Hill reaction activity was recorded more in PBW621 followed by C306 as compared with others. In all genotypes photolysis of water showed decline with increase in temperature stress. Trehalose pretreatment helped in sustaining Hill reaction activity probably by stabilizing the photosynthetic apparatus against heat-induced photo inhibition. Both plant growth and development were affected by temperature in both shoot and root under heat stress. The reduction was compensated partially by trehalose (1.5 mM) application. Adaption to heat stress is associated with the metabolic adjustment which led to accumulation of soluble sugars including non-reducing and reducing for their role in adaptive mechanism. Higher acid invertase activity in shoot of tolerant genotypes appeared to be a characteristic for stress tolerance. As sucrose synthase play central role in sink strength and in studied wheat genotype was positively related to dry matter accumulation. The duration of heat stress for eight hours had more severe effect on these parameters and trehalose application at 1.5 mM ameliorated it to certain extent.

Keywords: heat stress, Triticum aestivum, trehalose, membrane thermal stability, triphenyl tetrazolium chloride, reduction test, growth, sugar metabolism

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Authors: M. François, L. Sigot, C. Vallières

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Volatile Organic Compounds (VOCs) are a real health issue, particularly in domestic indoor environments. Among these VOCs, acetaldehyde is frequently monitored in dwellings ‘air, especially due to smoking and spontaneous emissions from the new wall and soil coverings. It is responsible for respiratory complaints and is classified as possibly carcinogenic to humans. Adsorption processes are commonly used to remove VOCs from the air. Metal-Organic Frameworks (MOFs) are a promising type of material for high adsorption performance. These hybrid porous materials composed of metal inorganic clusters and organic ligands are interesting thanks to their high porosity and surface area. The HKUST-1 (also referred to as MOF-199) is a copper-based MOF with the formula [Cu₃(BTC)₂(H₂O)₃]n (BTC = benzene-1,3,5-tricarboxylate) and exhibits unsaturated metal sites that can be attractive sites for adsorption. The objective of this study is to investigate the impact of HKUST-1 pretreatment on acetaldehyde adsorption. Thus, dynamic adsorption experiments were conducted in 1 cm diameter glass column packed with 2 cm MOF bed height. MOF were sieved to 630 µm - 1 mm. The feed gas (Co = 460 ppmv ± 5 ppmv) was obtained by diluting a 1000 ppmv acetaldehyde gas cylinder in air. The gas flow rate was set to 0.7 L/min (to guarantee a suitable linear velocity). Acetaldehyde concentration was monitored online by gas chromatography coupled with a flame ionization detector (GC-FID). Breakthrough curves must allow to understand the interactions between the MOF and the pollutant as well as the impact of the HKUST-1 humidity in the adsorption process. Consequently, different MOF water content conditions were tested, from a dry material with 7 % water content (dark blue color) to water saturated state with approximately 35 % water content (turquoise color). The rough material – without any pretreatment – containing 30 % water serves as a reference. First, conclusions can be drawn from the comparison of the evolution of the ratio of the column outlet concentration (C) on the inlet concentration (Co) as a function of time for different HKUST-1 pretreatments. The shape of the breakthrough curves is significantly different. The saturation of the rough material is slower (20 h to reach saturation) than that of the dried material (2 h). However, the breakthrough time defined for C/Co = 10 % appears earlier in the case of the rough material (0.75 h) compared to the dried HKUST-1 (1.4 h). Another notable difference is the shape of the curve before the breakthrough at 10 %. An abrupt increase of the outlet concentration is observed for the material with the lower humidity in comparison to a smooth increase for the rough material. Thus, the water content plays a significant role on the breakthrough kinetics. This study aims to understand what can explain the shape of the breakthrough curves associated to the pretreatments of HKUST-1 and which mechanisms take place in the adsorption process between the MOF, the pollutant, and the water.

Keywords: acetaldehyde, dynamic adsorption, HKUST-1, pretreatment influence

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Authors: Joseph A. Bentil, Anders Thygesen, Lene Langea, Moses Mensah, Anne Meyer

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The study investigated the saccharification potential of in-house enzymes produced from a white-rot basidiomycete strain, Polyporus ciliatus CBS 366.74. The in-house enzymes were produced by growing the fungus on mono and composite substrates of cocoa pod husk (CPH) and green seaweed (GS) (Ulva lactuca sp.) with and without the addition of 25mM ammonium nitrate at 4%w/v substrate concentration in submerged condition for 144 hours. The crude enzyme extracts preparations (CEE 1-5 and CEE 1-5+AN) obtained from the fungal cultivation process were sterile-filtered and used as enzyme sources for enzymatic hydrolysis of hydrothermally pretreated corn stover using a commercial cocktail enzyme, Cellic Ctec3, as benchmark. The hydrolysis was conducted at 50ᵒC with 50mM sodium acetate buffer, pH 5 based on enzyme dosages of 5 and 10 CMCase Units/g biomass at 1%w/v dry weight substrate concentration at time points of 6, 24, and 72 hours. The enzyme activity profile of the in-house enzymes varied among the growth substrates with the composite substrates (50-75% GS and AN inclusion), yielding better enzyme activities, especially endoglucanases (0.4-0.5U/mL), β-glucosidases (0.1-0.2 U/mL), and xylanases (3-10 U/mL). However, nitrogen supplementation had no significant effect on enzyme activities of crude extracts from 100% GS substituted substrates. From the enzymatic hydrolysis, it was observed that the in-house enzymes were capable of hydrolysing the pretreated corn stover at varying degrees; however, the saccharification yield was less than 10%. Consequently, theoretical glucose yield was ten times lower than Cellic Ctec3 at both dosage levels. There was no linear correlation between glucose yield and enzyme dosage for the in-house enzymes, unlike the benchmark enzyme. It is therefore recommended that the in-house enzymes are used to complement the dosage of commercial enzymes to reduce the cost of biomass saccharification.

Keywords: enzyme production, hydrolysis yield, feedstock, enzyme blend, Polyporus ciliatus

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Authors: Salwa Karboune, Amanda Waglay

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Recent studies indicate that health-promoting functional ingredients and nutraceuticals can help support and improve the overall public health, which is timely given the aging of the population and the increasing cost of health care. The development of novel ‘natural’ functional ingredients is increasingly challenging. Biocatalysis offers powerful approaches to achieve this goal. Our recent research has been focusing on the development of innovative biocatalytic approaches towards the isolation of protein isolates from potato by-products and the generation of peptides. Potato is a vegetable whose high-quality proteins are underestimated. In addition to their high proportion in the essential amino acids, potato proteins possess angiotensin-converting enzyme-inhibitory potency, an ability to reduce plasma triglycerides associated with a reduced risk of atherosclerosis, and stimulate the release of the appetite regulating hormone CCK. Potato proteins have long been considered not economically feasible due to the low protein content (27% dry matter) found in tuber (Solanum tuberosum). However, potatoes rank the second largest protein supplying crop grown per hectare following wheat. Potato proteins include patatin (40-45 kDa), protease inhibitors (5-25 kDa), and various high MW proteins. Non-destructive techniques for the extraction of proteins from potato pulp and for the generation of peptides are needed in order to minimize functional losses and enhance quality. A promising approach for isolating the potato proteins was developed, which involves the use of multi-enzymatic systems containing selected glycosyl hydrolase enzymes that synergistically work to open the plant cell wall network. This enzymatic approach is advantageous due to: (1) the use of milder reaction conditions, (2) the high selectivity and specificity of enzymes, (3) the low cost and (4) the ability to market natural ingredients. Another major benefit to this enzymatic approach is the elimination of a costly purification step; indeed, these multi-enzymatic systems have the ability to isolate proteins, while fractionating them due to their specificity and selectivity with minimal proteolytic activities. The isolated proteins were used for the enzymatic generation of active peptides. In addition, they were applied into a reduced gluten cookie formulation as consumers are putting a high demand for easy ready to eat snack foods, with high nutritional quality and limited to no gluten incorporation. The addition of potato protein significantly improved the textural hardness of reduced gluten cookies, more comparable to wheat flour alone. The presentation will focus on our recent ‘proof-of principle’ results illustrating the feasibility and the efficiency of new biocatalytic processes for the production of innovative functional food ingredients, from potato by-products, whose potential health benefits are increasingly being recognized.

Keywords: biocatalytic approaches, functional ingredients, potato proteins, peptides

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Authors: Sudkate Chaiyo, Weena Siangproh, Orawon Chailapakul, Kurt Kalcher

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In the present work, a simple and sensitive non-enzymatic electrochemical detection of glucose in disposable paper-based sensor was developed at ionic liquid/graphene/cobalt phthalocyanine composite (IL/G/CoPc) modified electrode. The morphology of the fabricated composite was characterized and confirmed by scanning electron microscopy and UV-Vis spectroscopy. The UV-Vis spectroscopy results confirmed that the G/CoPc composite formed via the strong π–π interaction between CoPc and G. Amperometric i-t technique was used for the determination of glucose. The response of glucose was linear over the concentration ranging from 10 µM to 1.5 mM. The response time of the sensor was found as 30 s with a limit of detection of 0.64 µM (S/N=3). The fabricated sensor also exhibited its good selectivity in the presence of common interfering species. In addition, the fabricated sensor exhibited its special advantages such as low working potential, good sensitivity along with good repeatability and reproducibility for the determination of glucose.

Keywords: glucose, paper-based sensor, ionic liquid/graphene/cobalt phthalocyanine composite, electrochemical detection

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Authors: Vanitha Varadharaj, Vijalakshmi Krishnamurthy

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Annona squamosa Linn, commonly known as Sugar apple, belonging to the family Annonaceae, is said to show varied medicinal effects, including insecticide, antiovulatory and abortifacient. The alkaloid and flavonoids present in Annona squamosa leaf has proved to have antioxidant activity. The present work has been planned to investigate the effect of ethanolic leaf extract of Annona squamosa leaf on Den Induced wistar albino rats. The study was carried out to analyze the biochemical Parmeters like Total Proteins, Bilirubin, Enzymatic and Non –Enzymatic enzymes, Marker enzymes and Tumor markers in serum and also the histopathological studies in liver is carried out in control and DEN induced rats. Supplementation of ELAS (Ethanolic Leaf Extract Of Annona squamosa) reduced the liver weight and also reduced the tumour incidence. Chemoprevention group showed near normal values of bilirubin when compared with the control rats. Total protein was decreased in the cancer bearing group and on treatment with the extract the levels of protein were restored. Both in pre and post treatment group, the activities of enzymatic antioxidants such as superoxide dismutase, catalase, and Glutathione peroxidase were increased but in pre treated animals it was more effective than post treated animals. The non- enzymatic antioxidants such as vitamin C and vitamin E were brought back to normal level significantly in post and pre treated animals. Activities of marker enzymes such as SGOT, SGPT, ALP, γ GT were significantly elevated in the serum of cancer animals and the values returned to normal after treatment with the extract suggesting the hepato protective effect of the extract. Lipid peroxide was found to be elevated in the cancer induced group. This condition was brought back to the normal in the pre and post treated animals with ELAS. Histological examination also confirmed the anti- carcinogenic potential of ELAS, Cancer induced groups had a triple fold increase in their AFP values when compared to other groups. DEN treatment increased the level of AFP expression while ELAS partially counteracted the effect of it. So the scientific validation obtained from this study may pave way to many budding scientists to find new drugs from Annona squamosa for various ailments.

Keywords: annona squamosa, biochemical parmeters, cancer, leaf extract

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Authors: Mohamed Bakry Mohamed Ali, Mohamed Ehab El-Din Mustafa, Joseph Naiem Sabet Aziz, Sarah Mahmoud Ali Kaooh

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Background: Acute lung injury (ALI) is one of the major challenges in intensive care medicine. The most common extrapulmonary cause of ALI is sepsis, accounting more than 30% of the cases in humans. Lipopolysaccharide (LPS) has gained wide acceptance as a clinically relevant model of ALI. Lipopolysaccharide is a glycoprotein forming the major constituent of bacterial endotoxin. Losartan is angiotensin II type 1 (AT1) receptor antagonists. It is widely used for management of hypertension. It was recently suggested that losartan protects against septic ALI. It would thereby prevent LPS-induced ALI. Aim of the work and design of the experiment: This work investigated the injurious effect of lipopolysaccharide (LPS) and ALI on adult male albino rat at 24 hours and 14 days of LPS administration and the possible protective role of losartan pretreatment. LPS has deteriorated animal survival and behavior. It increased lung weight and induced lung histological damage. These changes could be much reduced by the losartan pretreatment. Conclusion: Administration of losartan before LPS could largely reduce these LPS/ ALI induced short and long term alterations. It could be recommended that patients susceptible to developing ALI, as in ICU, should receive a protective dose of angitensin II type 1 (AT1) receptor blocker as losartan.

Keywords: acute lung injury (ALI), lipopolysaccharide (LPS), losartan

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Authors: Joyce Cruz Ferraz Dutra, Marcele Fonseca Passos, Rubens Maciel Filho, Douglas Fernandes Barbin, Gustavo Mockaitis

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The search for alternatives to control hunger in the world, generated a major environmental problem. Intensive systems of fish production can cause an imbalance in the aquatic environment, triggering the phenomenon of eutrophication. Currently, there are many forms of growth control aquatic plants, such as mechanical withdrawal, however some difficulties arise for their final destination. The Egeria densa is a species of submerged aquatic macrophyte-rich in cellulose and low concentrations of lignin. By applying the concept of second generation energy, which uses lignocellulose for energy production, the reuse of these aquatic macrophytes (Egeria densa) in the biofuels production can turn an interesting alternative. In order to make lignocellulose sugars available for effective fermentation, it is important to use pre-treatments in order to separate the components and modify the structure of the cellulose and thus facilitate the attack of the microorganisms responsible for the fermentation. Therefore, the objective of this research work was to evaluate the structural and morphological transformations occurring in the biomass of aquatic macrophytes (E.densa) submitted to a thermal pretreatment. The samples were collected in an intensive fish growing farm, in the low São Francisco dam, in the northeastern region of Brazil. After collection, the samples were dried in a 65 0C ventilation oven and milled in a 5mm micron knife mill. A duplicate assay was carried, comparing the in natural biomass with the pretreated biomass with heat (MT). The sample (MT) was submitted to an autoclave with a temperature of 1210C and a pressure of 1.1 atm, for 30 minutes. After this procedure, the biomass was characterized in terms of degree of crystallinity and morphology, using X-ray diffraction (XRD) techniques and scanning electron microscopy (SEM), respectively. The results showed that there was a decrease of 11% in the crystallinity index (% CI) of the pretreated biomass, leading to the structural modification in the cellulose and greater presence of amorphous structures. Increases in porosity and surface roughness of the samples were also observed. These results suggest that biomass may become more accessible to the hydrolytic enzymes of fermenting microorganisms. Therefore, the morphological transformations caused by the thermal pretreatment may be favorable for a subsequent fermentation and, consequently, a higher yield of biofuels. Thus, the use of thermally pretreated aquatic macrophytes (E.densa) can be an environmentally, financially and socially sustainable alternative. In addition, it represents a measure of control for the aquatic environment, which can generate income (biogas production) and maintenance of fish farming activities in local communities.

Keywords: aquatics macrophyte, biofuels, crystallinity, morphology, pretreatment thermal

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Authors: Yatimah Alias, Tilagam Marimuthu, M. R. Mahmoudian, Sharifah Mohamad

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The rapid growth of science and technology in energy and environmental fields has enlightened the substantial importance of the conducting polymer and metal composite materials engineered at nano-scale. In this study, polypyrrole-cobalt composites (PPy-Co Cs) and polypyrrole-nickel oxide composites (PPy-NiO Cs) were prepared by a simple and facile chemical polymerization method with an aqueous solution of pyrrole monomer in the presence of metal salt. These composites then fabricated into non-enzymatic hydrogen peroxide (H2O2) and glucose sensor. The morphology and composition of the composites are characterized by the Field Emission Scanning Electron Microscope, Fourier Transform Infrared Spectrum and X-ray Powder Diffraction. The obtained results were compared with the pure PPy and metal oxide particles. The structural and morphology properties of synthesized composites are different from those of pure PPy and metal oxide particles, which were attributed to the strong interaction between the PPy and the metal particles. Besides, a favorable micro-environment for the electrochemical oxidation of H2O2 and glucose was achieved on the modified glassy carbon electrode (GCE) coated with PPy-Co Cs and PPy-NiO Cs respectively, resulting in an enhanced amperometric response. Both PPy-Co/GCE and PPy-NiO/GCE give high response towards target analyte at optimum condition of 500 μl pyrrole monomer content. Furthermore, the presence of pyrrole monomer greatly increases the sensitivity of the respective modified electrode. The PPy-Co/GCE could detect H2O2 in a linear range of 20 μM to 80 mM with two linear segments (low and high concentration of H2O2) and the detection limit for both ranges is 2.05 μM and 19.64 μM, respectively. Besides, PPy-NiO/GCE exhibited good electrocatalytic behavior towards glucose oxidation in alkaline medium and could detect glucose in linear ranges of 0.01 mM to 0.50 mM and 1 mM to 20 mM with detection limit of 0.33 and 5.77 μM, respectively. The ease of modifying and the long-term stability of this sensor have made it superior to enzymatic sensors, which must kept in a critical environment.

Keywords: metal oxide, composite, non-enzymatic sensor, polypyrrole

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Authors: Hak-Ryul Kim, Hyung-Geun Lee, Qi Long, Ching Hou

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Structural modification of natural lipids via chemical reaction or microbial bioconversion can change their properties or even create novel functionalities. Enzymatic oxidation of lipids leading to formation of oxylipin is one of those modifications. Hydroxy fatty acids, one of those oxylipins have gained important attentions because of their structural and functional properties compared with other non-hydroxy fatty acids. Recently 7,10-dihydroxy-8(E)-octadecenoic acid (DOD) was produced with high yield from lipid-containing oleic acid by microbial conversion, and the further study confirmed that DOD contained strong antimicrobial activities against a broad range of microorganisms. In this study, we tried to modify DOD molecules by the enzymatic or physical reaction to create new functionality or to enhance the antimicrobial activity of DOD. After modification of DOD molecules by different ways, we confirmed that the antimicrobial activity of DOD was highly enhanced and presented strong antimicrobial activities against multidrug-resistant Staphylococcus aureus, suggesting that DOD and its derivatives can be used as efficient antimicrobial agents for medical and industrial applications.

Keywords: biocatalysis, antimicrobial agent, multidrug-resistant bacteria, vegetable oil

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Authors: SunWoo Lee, TaeBum Lee, YoonHwa Park, YooJeong Kim

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Solar lentigines appear predominantly on chronically sun-exposed areas of skin, such as the face and the back of the hands. Among the several ways to lentigines treatment, quality-switched lasers are well-known effective treatment for removing solar lentigines. The present pilot study was therefore designed to assess the efficacy of quality-switched ruby laser treatment of such lentigines compare between pretreatment and posttreatment of skin brightness. Twenty-two adults with chronic sun-damaged skin (mean age 52.8 years, range 37–74 years) were treated at the Korean site. A 694 nm Q-switched ruby laser was used, with the energy density set from 1.4 to 12.5 J/cm2, to treat solar lentigines. Average brightness of skin color before ruby laser treatment was 137.3 and its skin color was brightened after ruby laser treatment by 150.5. Also, standard deviation of skin color was decreased from 17.8 to 16.4. Regarding the multivariate model, age and energy were identified as significant factors for skin color brightness change in lentigo depigmentation by ruby laser treatment. Their respective odds ratios were 1.082 (95% CI, 1.007–1.163), and 1.431 (95% CI, 1.051–1.946). Lentigo depigmentation treatment using ruby lasers resulted in a high performance in skin color brightness. Among the relative factors involve with ruby laser treatment, age and energy were the most effective factors which skin color change to brighter than pretreatment.

Keywords: depigmentation, lentigine, quality switched ruby laser, skin color

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Authors: Fahimeh Helali Soltanahmadi, Alireza Nourabdi

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To investigate the effects of seed pretreatment on drought tolerance, an experimental Marigold (Calendula officinalis L) was tested in 2017-2018. A germination experiment was performed in the laboratory of Urmia University. The experiment was performed as a factorial experiment in a completely randomized design with three replications. The experimental treatments were dry levels of polyethylene glycol in four levels (control, -3, -6 and -9 bar) and treatments of Humic acid (72 mg / l), salicylic acid (2000 μmol) and ascorbic acid (200 ppm), and control were performed. The results of the germination experiment showed that with increasing drought levels caused by polyethylene glycol, germination indices were significantly reduced, so that the highest value obtained from the measured traits was observed in normal irrigation and in priming treatments, and the lowest amount of these traits was observed at the -9 bar drought level due to polyethylene glycol in the control treatment. The results of this study showed an increase in drought tolerance and improved Marigold germination under stress by applying seed priming with salicylic acid, which is recommended in sustainable and organic agriculture.

Keywords: Marigold, organic matter, seed pretreatment, drought stress, allometric coefficient

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Authors: Sonia Mbarki Marian Brestic, Artemio Cerda Naceur Jedidi, Jose Antonnio Pascual Chedly Abdelly

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Monitoring the effect addition bio-amendment as compost to an agricultural soil for growing plant lolium perenne irrigated with a CdCl2 solution at 50 µM on physicochemical soils characteristics and plant production in laboratory condition. Even microbial activity indexes (acid phosphatase, β-glucosidase, urease, and dehydrogenase) was determined. Basal respiration was the most affected index, while enzymatic activities and microbial biomass showed a decrease due to the cadmium treatments. We noticed that this clay soil with higher pH showed inhibition of basal respiration. Our results provide evidence for the importance of ameliorating effect compost on plant growth even when soil was added with cadmium solution at 50 µmoml.l-1. Soil heavy metal concentrations depended on heavy metals types, increased substantially with cadmium increase and with compost addition, but the recorded values were below the toxicity limits in soils and plants except for cadmium.

Keywords: compost, enzymatic activity, lolium perenne, bioremediation

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Authors: Muralisankar Thirunavukkarasu, Saravana Bhavan Periyakali, Santhanam Perumal

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The present study was performed to assess the effects of dietary copper (Cu) on growth, biochemical constituents, digestive enzyme activities, enzymatic antioxidant and metabolic enzymes of the freshwater prawn, Macrobrachium rosenbergii post larvae (PL). The Cu was supplemented at 0, 10, 20, 40, 60 and 80 mg kg-1 with the basal diets. Cu supplemented diets were fed to M. rosenbergii PL for a period of 90 days. At the end of the feeding experiment, 40 mg kg-1 Cu supplemented feeds fed PL showed significant (P < 0.05) improvement in survival, growth, digestive enzyme activities and concentrations of biochemical constituents. However, PL fed with 60 to 80 mg Cu kg-1 showed negative performance. Activities of enzymatic antioxidants, metabolic enzymes and lipid peroxidation in the muscle and hepatopancreas showed insignificant alterations (P > 0.05) up to 40 mg kg-1 Cu supplemented feeds fed PL. Whereas, 60 and 80 mg of Cu kg-1 supplemented feeds fed PL showed significant alterations on these antioxidants and metabolic enzymes levels. It indicates that beyond 40 mg Cu kg-1 diets were produced some toxic to M. rosenbergii PL. Therefore, the present study suggests that 40 mg Cu kg-1 can be supplemented in the diets of M. rosenbergii PL for regulating better survival and growth.

Keywords: antioxidants, biochemical constituents, copper, growth, Macrobrachium rosenbergii

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Authors: Kai-Shiang Chang, Shiao-Shing Chen, Saikat Sinha Ray, Hung-Te Hsu

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In recent years, membrane bioreactors (MBR) have been widely utilized as it can effectively replace conventional activated sludge process (CAS). Membrane bioreactor (MBR) is found to be more effective technology compared to other conventional activated sludge process and advanced membrane separation technique. Additionally, as far as the MBR is concerned, it is having excellent control of sludge retention time (SRT) and hydraulic retention time (HRT) and conducive to the retention of high concentration of sludge biomass. The membrane bioreactor (MBR) can effectively reduce footprint in terms of area and omit the secondary processing procedures in the conventional activated sludge process (CAS). Currently, as per the membrane technology, the ceramic membrane is found to have highly strong anti-acid-base properties, and it is more suitable than polymeric membrane while using for backwash and chemical cleaning. This study is based upon the treatment of Cutting Fluid wastewater, as the Cutting Fluid is widely used in the cutting equipment. However, the Cutting Fluid wastewater is very difficult to treat. In this study, the ceramic membrane was used and combine with of MBR system to treat the Cutting Fluid wastewater. In this present study, different kind of chemical coagulants have been utilized for pretreatment purpose in order to get the supernatant and simultaneously this wastewater (supernatant) was treated by MBR process. Nevertheless, ceramic membrane has three advantages such as high mechanical strength, drug resistance and reuse. During the experiment, the backwash technique was used for every interval of 10 minutes in order to avoid fouling of the membrane. In this study, during pretreatment the Chemical Oxygen Demand (COD) removal efficiency was found to be 71-86% and oil removal efficiency was analyzed to be 83-92%. This pretreatment study suggests that it is quiet effective methodology to reduce COD and oil concentration. Finally, In the MBR system when the HRT is more than 7.5 hour, the COD removal efficiency was found to be 87-93% and could achieve 100% oil removal efficiency. Coagulation test series were seen in Refs coagulants for the treatment of wastewater containing cutting oil with better oil and COD removal efficiency. The results also showed that the oil removal efficiency in the MBR system could reduce the oil content to less than 1 mg / L when the oil quality was 126 mg / L. Therefore, in this paper, the performance of membrane bioreactor by utilizing ceramic membrane has been demonstrated for treatment of Cutting Fluid wastewater.

Keywords: membrane bioreactor, cutting fluid, oil, chemical oxygen demand

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Authors: S. Thangapandiyan, S. Miltonprabu

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Fl (Fl) exposure engenders neurodegeneration and induces oxidative stress in the brain. The Neuroprotective role of EGCG on oxidative stress-mediated neurotoxicity in Fl intoxicated rat hippocampus has not yet been explored so far. Hence, the present study is focused on witnessing whether EGCG (40mg/kg) supplementation prevents Fl induced oxidative stress in the brain of rats with special emphasis on the hippocampus. Fl (25mg/kg) intoxication for four weeks in rats showed an increase in Fl concentration along with the decrease the AChE, NP, DA, and 5-HT activity in the brain. The oxidative stress markers (ROS, TBARS, NO, and PC) were significantly increased with decreased enzymatic (SOD, CAT, GPx, GR, GST, and G6PD) and non-enzymatic antioxidants (GSH, TSH, and Vit.C) in Fl intoxicated rat hippocampus. Moreover, Fl intoxicated rats exhibited an intrinsic and extrinsic pathway mediated apoptosis in the hippocampus of rats. Fl intoxication significantly increased the DNA damage as evidenced by increased DNA fragmentation. Furthermore, the toxic impact of Fl on hippocampus was also proved by the immunohistochemical, histological, and ultrastructural studies. Pre-administration of EGCG has significantly protected the Fl induced oxidative stress, biochemical changes, cellular apoptotic, and histological alternations in the hippocampus of rats. In conclusion, EGCG supplementation significantly attenuated the Fl induced oxidative stress mediated neurotoxicity via its free radical scavenging and antioxidant activity.

Keywords: brain, hippocampal, NaF, ROS, EGCG

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Authors: Friday A. Ogori, Ojotu M. Eke, Oneh J. Abu, Abraham T. Girgih

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B.aeqygtiaca del (Balanites aegyptiaca del) seed protein concentrate (APC) was hydrolyzed using different enzymes such as pepsin+pancreatin (PP), Alcalase (Alca), and Flavourzyme (Flav). The Alca has higher yield (100%) when compared to PP (83.23%) and Flav (62.90%). The hydrophobic amino acid and Sulphur containing amino acid (40.19%, 7.04%) in PP hydrolysate were higher compared to Alcalase (38.92%, 6.69%), Flavourenzyme (37.43%,6.35%), and APC (39.97%, 6.95%) samples. The PP has stronger DPPH, Hydroxyl radical quenching, Ferric reducing activity, and linoleic acid peroxidation activity, followed by the protein concentrate (APC) and Alcalase (Alca), while Flavourenzyme (Flav) derived hydrolysate was least in scavenging and inhibiting radical peroxidation properties. Flavourenzyme derived hydrolysate had stronger Ferric reducing antioxidant potential and metal chelating property. The result showed that the Alcalase hydrolysate has promising peptide yield, and PP hydrolysate had excellent amino acid residues and good in-vitro antioxidant potentials and could be a preferred ingredients in the nutraceutical and functional food emerging industries.

Keywords: balanites aegyptiaca del, protein concentrate, protein hydrolysates, enzymatic hydrolysis, antioxidants

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Authors: Parastoo Motallebi, Vahid Niknam, Hassan Ebrahimzadeh, Majid Hashemi

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Wheat, the most strategically important worldwide crop, is widely grown in various countries. Based on international wheat production statistics (FAOSTAT database), the total production of wheat in 2012 was 13.8 in Iran. Fusarium culmorum is one of the principal causative agents of Fusarium crown rot (FCR), an overwhelming disease of wheat and barley which is in the early stages causing yield losses, stand reductions and rotting of root and lower stem tissues. In this study inoculation of two wheat seedlings of the susceptible cultivar Falat and the partially field-resistant cultivar Pishtaz were carried out in greenhouse conditions and root samples were taken for 6 days. The activity of peroxidase (POX) and polyphenoloxidase (PPO) enzymes were analyzed to identify possible relations between resistance and enzymatic activities. Although the POX and PPO activities in both geno types increased, this significant increase was more dominant in Pishtaz. The results showed an earlier elevation in the activity of these defense related enzymes in semi-resistant cv. Pishtaz after inoculation, suggested that the activities of POX and PPO in wheat geno types play an important role in the induction of resistance to this disease.

Keywords: Defense responses, Fusarium culmorum, Wheat

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Authors: Mahmood Al Ramahi, G. Keszthelyi-Szabo, S. Beszedes

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Hydrothermal carbonization (HTC) is a thermochemical reaction that utilizes saturated water and vapor pressure to convert waste biomass to C-rich products This work evaluated the effect of HTC as a pre- and post-treatment technique to anaerobic digestion (AD) of dairy sludge, as information in this field is still in its infancy, with many research and methodological gaps. HTC effect was evaluated based on energy recovery, nutrients transformation, and sludge biodegradability. The first treatment approach was executed by applying hydrothermal carbonization (HTC) under a range of temperatures, prior to mesophilic anaerobic digestion (AD) of dairy sludge. Results suggested an optimal pretreatment temperature at 210 °C for 30 min. HTC pretreatment increased methane yield and chemical oxygen demand removal. The theoretical model based on Boyle’s equation had a very close match with the experimental results. On the other hand, applying HTC subsequent to AD increased total energy production, as additional energy yield was obtained by the solid fuel (hydrochar) beside the produced biogas. Furthermore, hydrothermal carbonization of AD digestate generated liquid products (HTC digestate) with improved chemical characteristics suggesting their use as liquid fertilizers.

Keywords: hydrothermal carbonization, anaerobic digestion, energy balance, sludge biodegradability, biogas

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Authors: Patsarawadee Paojinda, Waranya Imprasittichai, Sudaratana R. Krungkrai, Nirianne Marie Q. Palacpac, Toshihiro Horii, Jerapan Krungkrai

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Fusion of the last two enzymes in the pyrimidine biosynthetic pathway in the inversed order by having COOH-terminal orotate phosphoribosyltransferase (OPRT) and NH2-terminal orotidine 5'-monophosphate decarboxylase (OMPDC), as OMPDC-OPRT, are described in many organisms. Here, we produced gene fusions of Plasmodium falciparum OMPDC-OPRT and expressed the bifunctional protein in Escherichia coli. The enzyme was purified to homogeneity using affinity and anion-exchange chromatography, exhibited enzymatic activities and functioned as a dimer. The activities, although unstable, can be stabilized by its substrate and product during purification and long-term storage. Furthermore, the enzyme expressed a perfect catalytic efficiency (kcat/Km). The kcat was selectively enhanced up to 3 orders of magnitude, while the Km was not much affected and remained at low µM levels when compared to the monofunctional enzymes. The fusion of the two enzymes, creating a “super-enzyme” with perfect catalytic power and more flexibility, reflects cryptic relationship of enzymatic reactivaties and metabolic functions on molecular evolution.

Keywords: bifunctional enzyme, orotate phosphoribosyltransferase, orotidine 5'-monophosphate decarboxylase, plasmodium falciparum

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Authors: Masni Mat Yusoff, Michael H. Gordon, Keshavan Niranjan

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The study reports on the effect of aqueous enzymatic extraction (AEE) parameters on the Moringa oleifera (MO) oil yield and the formation of emulsion at the end of the process. A mixture of protease and cellulase enzymes was used at 3:1 (w/w) ratio. The highest oil yield of 19% (g oil/g sample) was recovered with the use of a mixture of pH 6, 1:4 material/moisture ratio, and incubation temperature, time, and shaking speed of 50 ⁰C, 12.5 hr, and 300 stroke/min, respectively. The use of pH 6 and 8 resulted in grain emulsions, while solid-intact emulsion was observed at pH 4. Upon fixing certain parameters, higher oil yield was extracted with the use of lower material/moisture ratio and higher shaking speed. Longer incubation time of 24 hr resulted in significantly (p < 0.05) similar oil yield with that of 12.5 hr, and an incubation temperature of 50 ⁰C resulted in significantly (p < 0.05) higher oil yield than that of 60 ⁰C. In overall, each AEE parameter showed significant effects on both the MO oil yields and the emulsions formed. One of the major disadvantages of an AEE process is the formation of emulsions which require further de-emulsification step for higher oil recovery. Therefore, critical studies on the effect of each AEE parameter may assist in minimizing the amount of emulsions formed whilst extracting highest total MO oil yield possible.

Keywords: enzyme, emulsion, Moringa oleifera, oil yield

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Authors: Kitti Szoke, Laszlo Szoke, Attila Czompa, Arpad Tosaki, Istvan Lekli

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Autophagy plays an important role in cardiac hypertrophy, which is one of the most common causes of heart failure in the world. This self-degradative catabolic process, responsible for protein quality control, balancing sources of energy at critical times, and elimination of damaged organelles. The autophagic activity can be triggered by starvation, oxidative stress, or pharmacological agents, like rapamycin. This induced autophagy can promote cell survival during starvation or pathological stress. In this study, it is investigated the effect of the induced autophagic process on angiotensin induced hypertrophic H9c2 cells. In our study, it is used H9c2 cells as an in vitro model. To induce hypertrophy, cells were treated with 10000 nM angiotensin-II, and to activate autophagy, 100 nM rapamycin treatment was used. The following groups were formed: 1: control, 2: 10000 nM AT-II, 3: 100 nM rapamycin, 4: 100 nM rapamycin pretreatment then 10000 nM AT-II. The cell viability was examined via MTT (cell proliferation assay) assay. The cells were stained with rhodamine-conjugated phalloidin and DAPI to visualize F-actin filaments and cell nuclei then the cell size alteration was examined in a fluorescence microscope. Furthermore, the expression levels of autophagic and apoptotic proteins such as Beclin-1, p62, LC3B-II, Cleaved Caspase-3 were evaluated by Western blot. MTT assay result suggests that the used pharmaceutical agents in the tested concentrations did not have a toxic effect; however, at group 3, a slight decrement was detected in cell viability. In response to AT-II treatment, a significant increase was detected in the cell size; cells became hypertrophic. However, rapamycin pretreatment slightly reduced the cell size compared to group 2. Western blot results showed that AT-II treatment-induced autophagy, because the increased expression of Beclin-1, p62, LC3B-II were observed. However, due to the incomplete autophagy, the apoptotic Cleaved Caspase-3 expression also increased. Rapamycin pretreatment up-regulated Beclin-1 and LC3B-II, down-regulated p62 and Cleaved Caspase-3, indicating that rapamycin-induced autophagy can restore the normal autophagic flux. Taken together, our results suggest that rapamycin activated autophagy reduces angiotensin-II induced hypertrophy.

Keywords: angiotensin-II, autophagy, H9c2 cell line, hypertrophy, rapamycin

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Authors: Yenny Paola Morales Cortés, Fabián Rico Rodríguez, Juan Carlos Serrato Bermúdez, Carlos Arturo Martínez Riascos

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Numerous benefits of galactooligosaccharides (GOS) as prebiotics have motivated the study of enzymatic processes for their production. These processes have special complexities due to several factors that make difficult high productivity, such as enzyme type, reaction medium pH, substrate concentrations and presence of inhibitors, among others. In the present work the production of galactooligosaccharides (with different degrees of polymerization: two, three and four) from lactose was studied. The study considers the formulation of a mathematical model that predicts the production of GOS from lactose using the enzyme β-galactosidase. The effect of pH in the reaction was studied. For that, phosphate buffer was used and with this was evaluated three pH values (6.0.6.5 and 7.0). Thus it was observed that at pH 6.0 the enzymatic activity insignificant. On the other hand, at pH 7.0 the enzymatic activity was approximately 27 times greater than at 6.5. The last result differs from previously reported results. Therefore, pH 7.0 was chosen as working pH. Additionally, the enzyme concentration was analyzed, which allowed observing that the effect of the concentration depends on the pH and the concentration was set for the following studies in 0.272 mM. Afterwards, experiments were performed varying the lactose concentration to evaluate its effects on the process and to generate the data for the adjustment of the mathematical model parameters. The mathematical model considers the reactions of lactose hydrolysis and transgalactosylation for the production of disaccharides and trisaccharides, with their inverse reactions. The production of tetrasaccharides was negligible and, because of that, it was not included in the model. The reaction was monitored by HPLC and for the quantitative analysis of the experimental data the Matlab programming language was used, including solvers for differential equations systems integration (ode15s) and nonlinear problems optimization (fminunc). The results confirm that the transgalactosylation and hydrolysis reactions are reversible, additionally inhibition by glucose and galactose is observed on the production of GOS. In relation to the production process of galactooligosaccharides, the results show that it is necessary to have high initial concentrations of lactose considering that favors the transgalactosylation reaction, while low concentrations favor hydrolysis reactions.

Keywords: β-galactosidase, galactooligosaccharides, inhibition, lactose, Matlab, modeling

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Authors: Ritu Chaturvedi, Manoj Paul

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A pot experiment was conducted to investigate the effect of Arbuscular mycorrhizal fungus (AMF) on metal(loid) uptake and accumulation efficiency of Pisum sativum along with physiological and biochemical response. Plants were grown in soil spiked with 50 and 100 mg kg-1 Pb, 25 and 50 mg kg-1 Cd, 50 and 100 mg kg-1 As and a combination of all three metal(loid)s. A parallel set was maintained and inoculated with arbuscular mycorrhizal fungus for comparison. After 60 days, plants were harvested and analysed for metal(loid) content. A steady increase in metal(loid) accumulation was observed on increment of metal(loid) dose and also on AMF inoculation. Plant height, biomass, chlorophyll, carotenoid and carbohydrate content reduced upon metal(loid) exposure. Increase in enzymatic (CAT, SOD and APX) and nonenzymatic (Proline) defence proteins was observed on metal(loid) exposure. AMF inoculation leads to an increase in plant height, biomass, chlorophyll, carotenoids, carbohydrate and enzymatic defence proteins (p≤0.001) under study; whereas proline content was reduced. Considering the accumulation efficiency and adaptive response of plants and alleviation of stress by AMF, this symbiosis can be applied for on-site remediation of Pb and Cd contaminated soil.

Keywords: heavy metal, mycorrhiza, pea, phyroremediation

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Authors: H. Merhni, A. Sbiti, I. Ratbi, A. Sefiani

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The proximal spinal muscular atrophy (SMA) is a group of neuromuscular disorders characterized by progressive muscle weakness due to the degeneration and loss of anterior motor neurons of the spinal cord. Depending on the age of onset of symptoms and their evolution, four types of SMA, varying in severity, result in a mutations of the SMN gene (survival of Motor neuron). We have analyzed the DNA of 295 patients referred to our genetic counseling; since January 1996 until October 2014; for suspected SMA. The homozygous deletion of exon 7 of the SMN gene was found in 133 patients; of which, 40.6% were born to consanguineous parents. In countries like Morocco, where the frequency of heterozygotes for SMA is high, genetic testing should be offered as first-line and, after careful clinical assessment, especially in newborns and infants with congenital hypotonia unexplained and prognosis compromise. The molecular diagnosis of SMA allows a quick and certainly diagnosis, provide adequate genetic counseling for families at risk and suggest, for couples who want prenatal diagnosis. The analysis of the SMN gene is a perfect example of genetic testing with an excellent cost/benefit ratio that can be of great interest in public health, especially in low-income countries. We emphasize in this work for the benefit of the generalization of molecular diagnosis of SMA by the technique of PCR-enzymatic digestion in other centers in Morocco.

Keywords: Exon7, PCR-digestion, SMA, SMN gene

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Authors: Mohammed Moustafa, Khaled Ayad, Waleed Reda

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Shoulder impingement syndrome is the most common disorder of the shoulder, resulting in functional loss and disability. Objective: This study was designed to compare between the effects of scapular muscle training versus rhythmic stabilization exercises in treatment of shoulder impingement syndrome. Methods: Thirty patients participated in this study; they were assigned randomly into two experimental groups. The first experimental group (A) consisted of 15 patients with a mean age (21.87±2.72) years; they received graduated rhythmic stabilization exercises and stretching of the posterior capsule. The second experimental group (B) consisted of 15 patients with a mean age (22.27±2.94) years; they received scapular muscle training exercises in addition to stretching of the posterior capsule. Treatment was given three times per week, every other day, for four consecutive weeks. Patients have been evaluated pretreatment and post treatment for shoulder pain severity and functional disability. Results: Both groups showed highly statistical significant reduction in pain severity and functional disability measured post-treatment when compared with their corresponding values in pretreatment assessment. Conclusion: Both of rhythmic stabilization exercises and scapular muscle training are effective interventions to reduce shoulder pain severity and functional disability.

Keywords: impingement syndrome, scapular exercises, rhythmic stabilization exercises, posterior capsule stretch

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Authors: Shivam Yadav, Neelam Atri

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Cyanobacteria, the photosynthetic prokaryotes are indispensable components of paddy soil contribute substantially to the nitrogen economy however often appended with metal load. They are well known to play crucial roles in maintenance of soil fertility and rice productivity. Nickel is one such metal that plays a vital role in the cellular physiology, however at higher concentrations it exerts adverse effects. Arsenic is another toxic metalloid that negatively affects the cyanobacterial proliferation. However species-specific comparative responses under As and Ni is largely unknown. The present study focuses on the comparative effects of nickel (Ni2+) and arsenite (As(III)) on two diazotrophic cyanobacterial species (Anabaena doliolum and Anabaena sp. PCC7120) in terms of antioxidative aspects. Oxidative damage measured in terms of lipid peroxidation and peroxide content was significantly higher after As(III) than Ni treatment as compared to control. Similarly, all the studied enzymatic and non-enzymatic parameters of antioxidative defense system except glutathione reductase (GR) showed greater induction against As(III) than Ni. Moreover, integrating comparative analysis of all studied parameters also demonstrated interspecies variation in terms of stress adaptive strategies reflected through higher sensitivity of Anabaena doliolum over Anabaena PCC7120.

Keywords: antioxidative system, arsenic, cyanobacteria, nickel

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Authors: Tami A. Ulhiza, Noor I. M. Puad, Azlin S. Azmi, Mohd. I. A. Malek

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High amount of chemical oxygen demand (COD) in starchy waste can be harmful to the environment. In common practice, starch processing wastewater is discharged to the river without proper treatment. However, starchy waste still contains complex sugars and organic acids. By the right pretreatment method, the complex sugar can be hydrolyzed into more readily digestible sugars which can be utilized to be converted into more valuable products. At the same time, the global demand of energy is inevitable. The continuous usage of fossil fuel as the main source of energy can lead to energy scarcity. Hydrogen is a renewable form of energy which can be an alternative energy in the future. Moreover, hydrogen is clean and carries the highest energy compared to other fuels. Biohydrogen produced from waste has significant advantages over chemical methods. One of the major problems in biohydrogen production is the raw material cost. The carbohydrate-rich starchy wastes such as tapioca, maize, wheat, potato, and sago wastes is a promising candidate to be used as a substrate in producing biohydrogen. The utilization of those wastes for biohydrogen production can provide cheap energy generation with simultaneous waste treatment. Therefore this paper aims to review variety source of starchy wastes that has been widely used to synthesize biohydrogen. The scope includes the source of waste, the performance in yielding hydrogen, the pretreatment method and the type of culture that is suitable for starchy waste.

Keywords: biohydrogen, dark fermentation, renewable energy, starchy waste

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Authors: Najwa Othman, Norhidayah Suleiman, Gun Hean Chong

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Palm fatty acid distillate (PFAD) is a by-product of palm oil refineries which contains valuable compounds such as phytosterols, squalene, polycosanol, co-enzyme Q10 and vitamin E (tocopherols and tocotrienols). Approximately 0.7-1.0% of vitamin E accumulates in PFAD, and it functions as antioxidants and anti-inflammatory. The objective of this research is to evaluate the effect of manipulated variables in supercritical carbon dioxide towards the recovery of tocotrienols in PFAD. The vitamin E concentrate isolated varies depending on the pre-treatment of sample and extraction techniques. In this research, tocotrienols in PFAD was concentrated by removing the extraneous matters, especially free fatty acid (FFA) and acylglycerols. Pre-treatment method such as enzymatic hydrolysis by using lipase from Candida rugosa as an enzyme was used to remove FFA and improve recovery of vitamin E. After that, treated PFAD was extracted by using supercritical fluid extraction in co-current glass beads packed column (22 cm x 75 cm i.d) at different temperatures (40-60°C) and pressures (100-300 bar) for 5 hours. After the extraction, the sample was analyzed by using high-pressure liquid chromatography (HPLC) system to quantify the tocotrienols. The results indicated that a combined pressure (200 bar) and temperature (60°C) was predicted to provide highest tocotrienols yield and the extraction yield obtained was 106.45%.

Keywords: enzymatic hydrolysis, palm fatty acid distillate, supercritical fluid extraction, tocotrienols

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