Search results for: anti-oxidant enzymes

Authors: Ferjani Hanen, El Arem Amira, Boussema Ayed Imen, Bacha Hassen

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Tacrolimus (TAC), a calcineurin inhibitor, is clinically used as an immunosuppressive agent in the transplant recipient, but its use associated-hepatotoxicity. Mycophenolate mofetil (MMF), an anti-metabolite, is a potent immunosuppressive drug. MMF is not hepatotoxic and is the most common adjunctive immunosuppressant for TAC. The effects of TAC and MMF combination in the liver is still not well understood. This work aimed to investigate their combined effect against in liver in rats Wistar after 24 h. The oral median lethal doses (LD50) of TAC and MMF alone were evaluated in rats are 240 mg/kg and 500 mg/kg respectively. Oral administration of the MMF at 50 mg/kg to male Wistar intoxicated with TAC at 60 mg/kg, demonstrated a significant protective effect by lowering the levels of hepatic markers enzymes (AST, ALT) in the serum rat. MMF attenuated oxidative stress by restoring the activities of SOD, CAT and by reducing the malondialdehyde (MDA) and protein carbonyl levels liver. This study provided evidence that MMF protects rat liver from TAC-induced injury and suggests a most combination use for organ transplantation.

Keywords: tacrolimus, mycophenolate mofetil, combination, liver, rat

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Authors: H. C. Ozdamar , O. Kilic-Erkek, H. E. Akkaya, E. Kilic-Toprak, M. Bor-Kucukatay

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Nordic hamstring exercise (NHE) is used to increase hamstring muscle strength, prevent injuries. The aim of this study was to reveal the acute, long-term effects of 10-week NHE, followed by 5, 10-week detraining on anthropometric measurements, flexibility, anaerobic power, muscle architecture, damage, fatigue, oxidative stress, plasma viscosity (PV), blood lactate levels. 40 sedentary, healthy male volunteers underwent 10 weeks of progressive NHE followed by 5, 10 weeks of detraining. Muscle architecture was determined by ultrasonography, stiffness by strain elastography. Anaerobic power was assessed by double-foot standing, long jump, vertical jump, flexibility by sit-lie, hamstring flexibility tests. Creatine kinase activity, oxidant/antioxidant parameters were measured from venous blood by a commercial kit, whereas PV was determined using a cone-plate viscometer. The blood lactate level was measured from the fingertip. NHE allowed subjects to lose weight, this effect was reversed by detraining for 5 weeks. Exercise caused an increase in knee angles measured by a goniometer, which wasn’t affected by detraining. 10-week NHE caused a partially reversed increase in anaerobic performance upon detraining. NHE resulted in increment of biceps femoris long head (BFub) area, pennation angle, which was reversed by detraining of 10-weeks. Blood lactate levels, muscle pain, fatigue were increased after each exercise session. NHE didn’t change oxidant/antioxidant parameters; 5-week detraining resulted in an increase in total oxidant capacity (TOC) and oxidative stress index (OSI). Detraining of 10 weeks caused a reduction of these parameters. Acute exercise caused a reduction in PV at 1 to 10 weeks. Pre-exercise PV measured on the 10th week was lower than the basal value. Detraining caused the increment of PV. The results may guide the selection of the exercise type to increase performance and muscle strength. Knowing how much of the gains will be lost after a period of detraining can contribute to raising awareness of the continuity of the exercise. This work was supported by PAU Scientific Research Projects Coordination Unit (Project number: 2018SABE034)

Keywords: anaerobic power, detraining, Nordic hamstring exercise, oxidative stress, plasma viscosity

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Authors: O. R. Raimi, A. Lateef

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Inoculum developments of A. niger were used for inoculation of medium for submerged fermentation and solid state fermentation. The filtrate obtained were used as sources of the extra-cellular enzymes. The FTase activities and the course of pH in submerged fermentation ranged from 7.53-24.42µ/ml and 4.4-4.8 respectively. The maximum FTase activity was obtained at 48 hours fermentation. In solid state fermentation, FTase activities ranged from 2.41-27.77µ/ml. Using ripe plantain peel and kola nut pod respectively. Both substrates supported the growth of the fungus, producing profuse growth during fermentation. In the control experiment (using kolanut pod) that lack supplementation, appreciable FTase activity of 16.92µ/ml was obtained. The optimum temperature range was 600C. it was also active at broad pH range of 1-9 with optimum obtain at pH of 5.0. FTase was stable within the range of investigated pH showing more than 60% activities. FTase can be used in the production of fructooligosaccharide, a functional food.

Keywords: Aspergillus niger, solid state fermentation, kola nut pods, Fructosyltransferase (FTase)

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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: Brian Chi Yan Cheng, Hui Guo, Tao Su, Xiu‐qiong Fu, Ting Li, Zhi‐ling Yu

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Rheumatoid arthritis (RA) affects around 1% of the globe population. Yet, there is still no cure for RA. Toll-like receptor 4 (TLR4) signalling has been found to be involved in the pathogenesis of RA, making it a potential therapeutic target for RA treatment. A herbal formula (RL) consisting of fruits of Rosa Multiflora (Eijitsu rose) and flowers of Lonicera Japonica (Japanese honeysuckle) has been used in treating various inflammatory disorders for more than a thousand year. Both of them are rich sources of nutrients and bioactive phytochemicals, which can be used in producing different food products and supplements. In this study, we would evaluate the anti-arthritic effect of RL on collagen-induced arthritis (CIA) in rats and investigate the involvement of TLR4 signaling in the mode of action of RL. Anti-arthritic efficacy was evaluated using CIA rats induced by bovine type II collagen. The treatment groups were treated with RL (82.5, 165, and 330 mg/kg bw per day, p.o.) or positive control indomethacin (0.25 mg/kg bw per day, p.o.) for 35 days. Clinical signs (hind paw volume and arthritis severity scores), changes in serum inflammatory mediators, pro-/antioxidant status, histological and radiographic changes of joints were investigated. Spleens and peritoneal macrophages were used to determine the effects of RL on innate and adaptive immune responses in CIA rats. The involvement of TLR4 signalling pathways in the anti-arthritic effect of RL was examined in cartilage tissue of CIA rats, murine RAW264.7 macrophages and human THP-1 monocytic cells. The severity of arthritis in the CIA rats was significantly attenuated by RL. Antioxidant status, histological score and radiographic score were efficiently improved by RL. RL could also dose-dependently inhibit pro-inflammatory cytokines in serum of CIA rats. RL significantly inhibited the production of various pro-inflammatory mediators, the expression and/or activity of the components of TLR4 signalling pathways in animal tissue and cell lines. RL possesses anti-arthritic effect on collagen-induced arthritis in rats. The therapeutic effect of RL may be related to its inhibition on pro-inflammatory cytokines in serum. The inhibition of the TAK1/NF-κB and TAK1/MAPK pathways participate in the anti-arthritic effects of RL. This provides a pharmacological justification for the dietary use of RL in the control of various arthritic diseases. Further investigation should be done to develop RL into a anti-arthritic food products and/or supplements.

Keywords: japanese honeysuckle, rheumatoid arthritis, rosa multiflora, rosehip

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Authors: Annesha Ghosh, S. B. Agrawal

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Agricultural sector is facing a serious threat due to climate change and exacerbation of different atmospheric pollutants. Tropospheric ozone (O₃) is considered as a dynamic air pollutant imposing substantial phytotoxicity to natural vegetations and agriculture worldwide. Naturally, plants are exposed to different environmental factors and their interactions. Amongst such interactions, studies related to O₃ and water stress are still rare. In the present experiment, wheat cultivar HD2967 were grown in open top chambers (OTC) under two O₃ concentration; ambient O₃ level (A) and elevated O₃ (E) (ambient + 20 ppb O₃) along with two different water supply; well-watered (W) and 50% water stress conditions (WS), with an aim to assess the individual and interactive effect of two most prevailing stress factors in Indo-Gangetic Plains of India. Exposure to elevated O₃ dose caused early senescence symptoms and reduction in growth and biomass of the test cultivar. The adversity was more pronounced under the combined effect of EWS. Significant reduction of stomatal conductance (gs) and assimilation rate were observed under combined stress condition compared to the control (AW). However, plants grown under individual stress conditions displayed higher gs, biomass, and antioxidant defense mechanism compared to the plants grown under the presence of combined stresses. Higher induction in most of the enzyme activities of catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), peroxidase (POD) and superoxide dismutase (SOD) was displayed by HD 2967 under EW while, under the presence of combined stresses (EWS), a moderate increment of APX and CAT activity was observed only at its vegetative phase. Furthermore, variations in nutrient uptake and redistribution to different plants parts were also observed in the present study. Reduction in water availability has checked nutrient uptake (N, K, P, Ca, Cu, Mg, Zn) in above-ground parts (leaf) and below-ground parts (root). On the other hand, carbon (C) accumulation with subsequent C-N ratio was observed to be higher in the leaves under EWS. Such major nutrient check and limitation in carbon fixation due to lower gs under combined stress conditions might have weakened the defense mechanisms of the test cultivar. Grain yield was significantly reduced under EWS followed by AWS and EW as compared to their control, exhibiting an additive effect on the grain yield.

Keywords: antioxidants, open-top chambers, ozone, water stress, wheat, yield

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Authors: Hailemeleak Regassa

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Traditional medicines use medicinal plants as a source of ingredients, and many modern medications are indirectly derived from plants. Consumers in affluent nations are growing disenchanted with contemporary healthcare and looking for alternatives. Oxidative stress is the primary cause of multiple diseases, and exogenous antioxidant supplementation or strengthening the body's endogenous antioxidant defenses are potential ways to counteract the negative effects of oxidative damage. Plants can biosynthesize non-enzymatic antioxidants that can reduce ROS-induced oxidative damage. Aging often aids the propagation and development of carcinogenesis, and older animals and older people exhibit increased vulnerability to tumor promoters. Cancer is a major public health issue, with several anti-cancer medications in clinical use. Potential drugs such as flavopiridol, roscovitine, combretastatin A-4, betulinic acid, and silvestrol are in the clinical or preclinical stages of research. Methodology: Microbial Growth media, Dimethyl sulfoxide (DMSO), methanol, ethyl acetate, and n-hexane were obtained from Himedia Labs, Mumbai, India. plant were collected from the Herbal Garden of Shoolini University campus, Solan, India (Latitude - 30.8644° N and longitude - 77.1184° E). The identity was confirmed by Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni, Solan (H.P.), India, and documented in Voucher specimens - UHF- Herbarium no. 13784; vide book no. 3818 Receipt No. 086. The plant materials were washed with tap water, and 0.1% mercury chloride for 2 minutes, rinsed with distilled water, air dried, and kept in a hot air oven at 40ºc on blotting paper until all the water evaporated and became well dried for grinding. After drying, the plant materials were grounded using a mixer grinder into fine powder transferred into airtight containers with proper labeling, and stored at 4ºc for future use (Horablaga et al., 2023). The extraction process was done according to Altemimi et al., 2017. The 5g powder was mixed with 15 ml of the respective solvents (n-hexane, ethyl acetate, and methanol), and kept for 4-5 days on the platform shaker. The solvents used are based on their increasing polarity index. Then the extract was centrifuged at 10,000rpm for 5 minutes and filtered using No.1 Whatman filter paper.

Keywords: cancer, phytomedicine, medicinal plants, oncology

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Authors: Abdelaaty A. Shahat, Mansour S. Alsaid

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Four Acetylated flavonol glycosides were isolated from Centaurium spicatum (L.) Fritsch (Gentianaceae). Structure elucidation, especially the localization of the acetyl groups, and complete 1H and 13C NMR assignments of these biologically active compounds were carried out using one- and two-dimensional NMR methods, including CNMR, DEPT-135 and DEPT-90 and gradient-assisted experiments such as DQF-COSY, TOCSY, HSQC and HMBC experiments. The antioxidant activities of the new acetylated flavonoid glycosides using DPPH• assay were determined. The compounds tested showed a good DPPH• activity compared with control, but their activity was lower than that of their corresponding aglycone, quercetin.

Keywords: Centaurium spicatum, flavonoids, biological activity, isolation, glycosides

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Authors: Shahram Nanekarania, Majid Goodarzia

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Calpastatin and callipyge have been known as one of the candidate genes in meat quality and quantity. Calpastatin gene has been located to chromosome 5 of sheep and callipyge gene has been localized in the telomeric region on ovine chromosome 18. The objective of this study was identification of calpastatin and callipyge genes polymorphism and analysis of genotype structure in population of Lori sheep kept in Iran. Blood samples were taken from 120 Lori sheep breed and genomic DNA was extracted by salting out method. Polymorphism was identified using the PCR-RFLP technique. The PCR products were digested with MspI and FaqI restriction enzymes for calpastatin gene and callipyge gene, respectively. In this population, three patterns were observed and AA, AB, BB genotype have been identified with the 0.32, 0.63, 0.05 frequencies for calpastatin gene. The results obtained for the callipyge gene revealed that only the wild-type allele A was observed, indicating that only genotype AA was present in the population under consideration.

Keywords: polymorphism, calpastatin, callipyge, PCR-RFLP, Lori sheep

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Authors: Serap Pektas

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Ataxia telangiectasia mutated (ATM) is a serine-threonine kinase, which is the major regulator of the DNA damage response. ATM is activated upon the formation of DNA double-strand breaks (DSBs) in the cells. ATM phosphorylates the proteins involved in apoptotic responses, cell cycle checkpoint control, DNA repair, etc. Tumor protein p53, known as p53 is one of these proteins that phosphorylated by ATM. Phosphorylation of p53 at Ser15 residue leads to p53 stabilization in the cells. Often enzymes activity is affected by hydrogen ion concentration (pH). In order to find the optimal pH range for ATM activity, steady-state kinetic assays were performed at acidic and basic pH ranges. Ser15 phosphorylation of p53 is determined by using ELISA. The results indicated that the phosphorylation rate was better at basic pH range compared with the acidic pH range. This could be due to enzyme stability, or enzyme-substrate interaction is pH dependent.

Keywords: ataxia telangiectasia mutated, DNA double strand breaks, DNA repair, tumor protein p53

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Authors: S. Buloyan, V. Mamikonyan, H. Hakobyan, H. Harutyunyan, H. Gasparyan

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The liver is the strongest regenerating organ of the organism, and even with 2/3 surgically removed, it can regenerate completely. Hence, liver cirrhosis may only develop when the regenerating system is off. We present the results of a comparative study of structural and functional characteristics of rat liver tissue under the conditions of toxic liver cirrhosis development, induced by carbon tetrachloride, and its prevention/treatment by natural compounds with antioxidant and immune stimulating action. Studies were made on Wister rats, weighing 120~140 g. Grape seeds extracts, separately and in combination with well known anticirrhotic drug ursodeoxycholic acid (ursodiol) have demonstrated effectiveness in prevention of liver cirrhosis development and its treatment.

Keywords: carbon tetrachloride, GSE, liver cirrhosis, prevention, treatment

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Authors: Ahmed F. Ghanem, Marwa I. Wahba, Asmaa N. El-Dein, Mohamed A. EL-Raey, Ghada E. A. Awad

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Numerous attempts are being performed in order to formulate suitable packaging materials for the meat products. However, to the best of our knowledge, the incorporation of the free hibiscus extract or its microcapsules in the pure polyvinyl alcohol (PVA) matrix as packaging materials for the meats is seldom reported. Therefore, this study aims at the protection of the aqueous crude extract of the hibiscus flowers utilizing the spry drying encapsulation technique. Results of the Fourier transform infrared (FTIR), the scanning electron microscope (SEM), and the particle size analyzer confirmed the successful formation of the assembled capsules via strong interactions, the spherical rough microparticles, and the particle size of ~ 235 nm, respectively. Also, the obtained microcapsules enjoy higher thermal stability than the free extract. Then, the obtained spray-dried particles were incorporated into the casting solution of the pure PVA film with a concentration of 10 wt. %. The segregated free-standing composite films were investigated, compared to the neat matrix, with several characterization techniques such as FTIR, SEM, thermal gravimetric analysis (TGA), mechanical tester, contact angle, water vapor permeability, and oxygen transmission. The results demonstrated variations in the physicochemical properties of the PVA film after the inclusion of the free and the extract microcapsules. Moreover, biological studies emphasized the biocidal potential of the hybrid films against the microorganisms contaminating the meat. Specifically, the microcapsules imparted not only antimicrobial but also antioxidant activities to the PVA matrix. Application of the prepared films on the real meat samples displayed a low bacterial growth with a slight increase in the pH over the storage time which continued up to 10 days at 4 oC, as further evidence to the meat safety. Moreover, the colors of the films did not significantly changed except after 21 days indicating the spoilage of the meat samples. No doubt, the dual-functional of the prepared composite films pave the way towards combined active and smart food packaging applications. This would play a vital role in the food hygiene, including also the quality control and the assurance.

Keywords: PVA, hibiscus, extraction, encapsulation, active packaging, smart and intelligent packaging, meat spoilage

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Authors: Tasleem Zafar, Jiwan Sidhu

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Introduction: Chickpea flour is known to offer many benefits to diabetic persons, especially in maintaining their blood sugar levels in the acceptable range. Under this project we have studied the chemical composition and antioxidant capacity of white flour (WF), whole wheat flour (WWF) and chickpea flour (BF), in addition to the effect of replacement of WF and WWF with BF on the rheological characteristics of these flour blends, with the ultimate objective of producing acceptable quality flat as well as pan-bread for the diabetic consumers. Methods: WF and WWF were replaced with BF ranging from 0 to 40%, to investigate its effect on the rheological properties and functionality of blended flour dough using farinograph, viscoamylograph, mixograph and falling number apparatus as per the AACC standard methods. Texture Profile Analysis (TPA) was carried on the WF, WWF, and their blends with BF using Stable Micro System Texture Analyzer. Effect of certain additives, such as freeze-dried amla fruit powder (Phyllanthus emblica L.), guar gum, and xanthan gum on the dough rheological properties were also studied. Results: Freeze-dried amla fruit powder was found to be very rich in ascorbic acid and other phenolics having higher antioxidant activity. A decreased farinograph water absorption, increased dough development time, higher mixing tolerance index (i.e., weakening of dough), decreased resistance to extension, lower ratio numbers were obtained when the replacement with BF was increased from 0 to 40%. The BF gave lower peak viscosity, lower paste breakdown, and lower setback values when compared with WF. The falling number values were significantly lower in WWF (meaning higher α-amylase activity) than both the WF and BF. Texture Profile Analysis (TPA) carried on the WF, WWF, and their blends with BF showed significant variations in hardness and compressibility values, dough becoming less hard and less compressible when the replacement of WF and WWF with BF was increased from 0 to 40%. Conclusions: To overcome the deleterious effects of adding BF to WF and WWF on the rheological properties will be an interesting challenge when good quality pan bread and Arabic flatbread have to be commercially produced in a bakery. Use of freeze-dried amla fruit powder, guar gum, and xanthan gum did show some promise to improve the mixing characteristics of WF, WWF, and their blends with BF, and these additives are expected to be useful in producing an acceptable quality flat as well as pan-bread on a commercial scale.

Keywords: wheat flour, chickpea flour, amla fruit, rheology

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Authors: Angel Champion

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Chemotherapy and radiation use an acidified approach to induce apoptosis, which only kills mature cancer cells while resulting in gene and cell damage with significant levels of toxicity in tumor-affected tissues and organs. The acid approach, where the cells exterminated are not differentiated, induces the disappearance of white blood cells from the blood. This increases susceptibility to infection in severe forms of cancer spread. However, chemotherapy and radiation cannot kill cancer stem cells that metastasize, being the leading cause of 98% of cancer fatalities. With over 12 million new cancer cases symptomatic each year, including common malignancies such as Hepatocellular Carcinoma (HCC), this study aims to assess the bioactive constituents and phytochemical composition of Taraxacum Officinale (Dandelion). This analysis enables pharmaceutical quality and potency to be applied to studies on cancer cell proliferation and apoptosis. A phytochemical screening is carried out to identify the antioxidant components of Dandelion root, stem, and flower extract. The constituents tested for are phlorotannins, carbohydrates, glycosides, saponins, flavonoids, alkaloids, sterols, triterpenes, and anthraquinone glycosides. To conserve the existing phenolic compounds, a portion of the constituent tests will be examined with an acid, alcohol, or aqueous solvent. As a result, the qualitative and quantitative variations within the Dandelion extract that measure uniform effective potency are vital to the conformity for producing medicinal products. These medicines will be constructed with a consistent, uniform composition that physicians can use to control and effectively eradicate malignant diseases safely. Taraxacum Officinale's phytochemical composition comprises a highly-graded potency due to present bioactive contents that will essentially drive out malignant disease within the human body. Its high potency rate is powerful enough to eliminate both mature cancer cells and cancer stem cells without the cell and gene damage induced by chemotherapy and radiation. Correspondingly, the high margins of cancer mortality on a global scale are mitigated. This remarkable contribution to modern therapeutics will essentially optimize the margins of natural products and their derivatives, which account for 50% of pharmaceuticals in modern therapeutics, while preventing the adverse effects of radiation and chemotherapy drugs.

Keywords: antioxidant, apoptosis, metastasize, phytochemical, proliferation, potency

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Authors: Raymond Oriebe Anyasi, Harrison Atagana

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The use of a plant in the detoxification of several toxin is been known to be enhanced by various microbial endophytes which have been reported to be contained in plants growing in any contaminated soil. Plants in their natural state are mostly colonized by endophytes which in the process forms symbiotic associations with the host plants. These benefits that the endophytes offer to the plants include amongst others to: Enhance plants growth through the production of various phytohormones; increase in the resistance of environmental stresses; produce important bioactive metabolites; help in the fixing of nitrogen in the plants organelles; help in the metal translocation and accumulation in plants; assist in the production of enzymes involves the degradation of organic contaminants. Therefore recognizing these natural processes of the microbes will enable the understanding of the effective mechanism for enhanced phytoremediation. The aim of this study was to survey the progressiveness in the study involving endophyte-assisted phytoremediation of contaminants; highlighting various pollutants, the plants used, the endophytes studied as well as the type of interaction between the plants and the microbes so as to proffer a better future prospect for the technology.

Keywords: phytoremediation, endophytes, microbes, pollution, environmental management, plants

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Authors: Demetrio L. Valle Jr., Juliana Janet M. Puzon, Windell L. Rivera

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From the various medicinal plants available in the Philippines, crude ethanol extracts of twelve (12) Philippine medicinal plants, namely: Senna alata L. Roxb. (akapulko), Psidium guajava L. (bayabas), Piper betle L. (ikmo), Vitex negundo L. (lagundi), Mitrephora lanotan (Blanco) Merr. (Lanotan), Zingiber officinale Roscoe (luya), Curcuma longa L. (Luyang dilaw), Tinospora rumphii Boerl (Makabuhay), Moringga oleifera Lam. (malunggay), Phyllanthus niruri L. (sampa-sampalukan), Centella asiatica (L.) Urban (takip kuhol), and Carmona retusa (Vahl) Masam (tsaang gubat) were studied. In vitro methods of evaluation against selected Gram-positive and Gram-negative multidrug-resistant (MDR), bacteria were performed on the plant extracts. Although five of the plants showed varying antagonistic activities against the test organisms, only Piper betle L. exhibited significant activities against both Gram-negative and Gram-positive multidrug-resistant bacteria, exhibiting wide zones of growth inhibition in the disk diffusion assay, and with the lowest concentrations of the extract required to inhibit the growth of the bacteria, as supported by the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Further antibacterial studies of the Piper betle L. leaf, obtained by three extraction methods (ethanol, methanol, supercritical CO2), revealed similar inhibitory activities against a multitude of Gram-positive and Gram-negative MDR bacteria. Thin layer chromatography (TLC) assay of the leaf extract revealed a maximum of eight compounds with Rf values of 0.92, 0.86, 0.76, 0.53, 0.40, 0.25, 0.13, and 0.013, best visualized when inspected under UV-366 nm. TLC- agar overlay bioautography of the isolated compounds showed the compounds with Rf values of 0.86 and 0.13 having inhibitory activities against Gram-positive MDR bacteria (MRSA and VRE). The compound with an Rf value of 0.86 also possesses inhibitory activity against Gram-negative MDR bacteria (CRE Klebsiella pneumoniae and MBL Acinetobacter baumannii). Gas Chromatography-Mass Spectrometry (GC-MS) was able to identify six volatile compounds, four of which are new compounds that have not been mentioned in the medical literature. The chemical compounds isolated include 4-(2-propenyl)phenol and eugenol; and the new four compounds were ethyl diazoacetate, tris(trifluoromethyl)phosphine, heptafluorobutyrate, and 3-fluoro-2-propynenitrite. Phytochemical screening and investigation of its antioxidant, cytotoxic, possible hemolytic activities, and mechanisms of antibacterial activity were also done. The results showed that the local variant of Piper betle leaf extract possesses significant antioxidant, anti-cancer and antimicrobial properties, attributed to the presence of bioactive compounds, particularly of flavonoids (condensed tannin, leucoanthocyanin, gamma benzopyrone), anthraquinones, steroids/triterpenes and 2-deoxysugars. Piper betle L. is also traditionally known to enhance wound healing, which could be primarily due to its antioxidant, anti-inflammatory and antimicrobial activities. In vivo studies on mice using 2.5% and 5% of the ethanol leaf extract cream formulations in the excised wound models significantly increased the process of wound healing in the mice subjects, the results and values of which are at par with the current antibacterial cream (Mupirocin). From the results of the series of studies, we have definitely proven the value of Piper betle L. as a source of bioactive compounds that could be developed into therapeutic agents against MDR bacteria.

Keywords: Philippine herbal medicine, multidrug-resistant bacteria, Piper betle, TLC-bioautography

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Authors: Yuli Andrea P. Bermudez, Richard R. Lobo, Tamyres R. D. Amorim, Danny Alexander R. Moreno, Angelica Simone C. Pereira, Ives Claudio D. Bueno

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The use of natural antioxidants in animal feed can positively modify the profile of fatty acids (FAs) in meat, due to the presence of secondary metabolites, mainly phenolic and flavonoid compounds, which promote an increase in the associated polyunsaturated fatty acids (PUFA) with beneficial factors in human health. The goal of this study was to evaluate the effect of the dietary inclusion percentage of yerba mate extract (Ilex paraguariensis St. Hilaire) as a natural antioxidant on lamb meat quality. The animals were confined for 53 days and fed with corn silage and concentrated in the proportion of 60:40, respectively, were divided into four homogeneous groups (n = 9 lambs/group), to each of the treatments, one control group without yerba mate extract - YME (0%) and three treatments with 1, 2 and 4% the inclusion of YME on a DM basis. Samples of the Longissimus thoracis (LT) muscle were collected from the deboning of 36 lambs, analyzing pH values, color parameters (brightness: L*, red value: a*, and yellow: b*), fatty acid profile, total lipids, and sensory analysis. The inclusion of YME modified the value of b* (P = 0.0041), indicating a higher value of yellow color in the meat, for the group supplemented with 4% YME. All data were statistically evaluated using the MIXED procedure of the statistical package SAS 9.4. However, it did not show differences in the final live weight in the groups evaluated, as well as in the pH values (P = 0.1923) and the total lipid concentration (P = 0.0752). The FAs (P ≥ 0.1360) and health indexes were not altered by the inclusion of YME (P ≥ 0.1360); only branched-chain fatty acids (BCFA) exhibited a diet effect (P = 0.0092) in the group that had 4% of the extract. In the sensory analysis test with a hedonic scale it did not show differences between the treatments (P ≥ 0.1251). Nevertheless, in the just about-right test, using (note 1) to 'very strong, softness or moist' (note 5); the softness was different between the evaluated treatments (P = 0.0088) where groups with 2% YME had a better acceptance of tasters (4.15 ± 0.08) compared to the control (3.89 ± 0.08). In conclusion, although the addition of YME has shown positive results in sensory acceptance and in increasing the concentration of BCFA, fatty acids beneficial to human health, without changing the physical-chemical parameters in lamb meat, the absolute changes are considered to have been quite small, which was probably related to the high efficiency of PUFA biohydrogenation in the n the rumen.

Keywords: composition, health, antioxidant, meat analysis

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Authors: Aliyu Adamu, Fahrul Huyop, Roswanira Abdul Wahab, Mohd Shahir Shamsir

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Halogenated organic compounds occur in huge amount in biosphere. This is attributable to the diverse use of halogen-based compounds in the synthesis of various industrially important products. Halogenated compound is toxic and may persist in the environment, thereby causing serious health and environmental pollution problems. L-2-haloacid dehalogenases (EC 3.8.1.2) catalyse the specific cleavage of carbon-halogen bond in L-isomers of halogenated compounds, which consequently reverse the effects of environmental halogen-associated pollution. To enhance the efficiency and utility of these enzymes, this study investigates the catalytic amino acid residues and the molecular functional mechanism of DehL, by classical molecular dynamic simulations, MM-PBSA and ab initio fragments molecular orbital (FMO) calculations. The results of the study will serve as the basis for the molecular engineering of the enzyme.

Keywords: DehL, Functional mechanism, Catalytic residues, L-2-haloacid dehalogenase

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Authors: Aisha Musaazi Sebunya Nakitto, Anika E. Wagner, Yusuf B. Byaruhanga, John H. Muyonga

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The prevalence and burden of diabetes are rapidly increasing globally, stemming from changes in lifestyle and dietary habits. Although several drugs are available to treat type 2 diabetes mellitus (T2DM), many are accompanied by several side effects and are often costly. Solanum anguivi Lam. fruits (SALF) are bitter berries that commonly grow in the wild and are traditionally cultivated by many globally as a remedy for T2DM. This effect is likely attributable to the presence of bioactive compounds such as phenolics, flavonoids, saponins, alkaloids, and vitamin C in SALF. In this study, we investigated the morphological characteristics of different SALF accessions, and the effect of ripeness stages and thermal treatments on the bioactive compounds contents (BCC) and antioxidant activity (AA) of SALF accessions. Using the fruit fly, Drosophila melanogaster (D. melanogaster) model, we explored the potential impact of dietary SALF in preventing and treating T2DM phenotypes. Morphological characterization was conducted based on descriptors of Solanum species. The BCC and AA of SALF at different ripeness stages (unripe, yellow, orange, and red) and after thermal treatments were determined using spectrophotometry, HPLC, and gravimetry. Male and female fruit flies were fed a high-sugar diet (HSD) to induce a T2DM-like phenotype, while control flies were fed on SY10 medium upto 24 days. Experimental flies were exposed to HSD supplemented with 5 or 10 mg/ml SALF. The therapeutic and prevention effect of SALF in T2DM-like phenotype were investigated on weight, climbing activity, glucose and triglyceride contents, survival, and gene expression of PPARγ co-activator 1α fly homolog Srl and Drosophila insulin-like peptides. Methods in fly studies included Gustatory assay, Climbing assay, Glucose GOD-PAP assay, Triglyceride GPO-PAP assay, Roti-Quant®, and Real Time-PCR analysis. Ripeness stage significantly influenced SALF BCC and AA, and this was dependent on the accession. The unripe stage had the highest AA and total phenolics and flavonoids; orange stage was rich in saponins, while the red stage had the highest alkaloid contents. Boiling and steaming increased the total phenolics and AA upto 4-fold and 3-fold, respectively. Drying at low temperature resulted in higher phenolics and AA than the control. In the therapeutic model, the HSD-fed female flies exhibited elevated glucose levels, which exhibited a dose-dependent reduction upon exposure to SALF-supplemented diet. Female flies fed on a SALF+ HSD exhibited a significant increase in survival compared to HSD-fed and control diet-fed flies. SALF supplementation did not alter weights, fitness, and triglyceride levels of female flies in comparison with HSD-only-fed flies. The mRNA levels of Srl decreased in HSD-fed flies compared to the control-fed, with no effect observed in females exposed to HSD+SALF. Similarly, in the preventative model, SALF diet resulted in higher survival of supplemented flies compared to controls. Consumption of boiled unripe SALF may result in the highest health benefits due to high phenolic contents and antioxidant activity observed. Dietary intake of SALF significantly lowered glucose levels and increased survival of the D. melanogaster model. Additional studies in higher organisms are needed to explore the preventative and therapeutic potential of SALF in T2DM.

Keywords: solanum anguivi, type 2 diabetes mellitus, bioactive compounds, drosophila melanogaster, survival, antioxidant activity

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Authors: Ahmad Manbohi, Seyyed Hamid Ahmadi

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A low-cost paper-based microfluidic device (PAD) for the multiplex electrochemical determination of glucose, uric acid, and dopamine in biological fluids was developed. Using wax printing, PAD containing a central zone, six channels, and six detection zones was fabricated, and the electrodes were printed on detection zones using pre-made electrodes template. For each analyte, two detection zones were used. The carbon working electrode was coated with chitosan-BSA (and enzymes for glucose and uric acid). To detect glucose and uric acid, enzymatic reactions were employed. These reactions involve enzyme-catalyzed redox reactions of the analytes and produce free electrons for electrochemical measurement. Calibration curves were linear (R^² > 0.980) in the range of 0-80 mM for glucose, 0.09–0.9 mM for dopamine, and 0–50 mM for uric acid, respectively. Blood samples were successfully analyzed by the proposed method.

Keywords: biological fluids, biomarkers, microfluidic paper-based electrochemical biosensors, Multiplex

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Authors: Anuranjani Kumar, Madhu Bala

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Ionising radiation exposure induces generation of free radicals and the oxidative DNA damage. SBL-1, a radioprotective leaf extract prepared from leaves Hippophae rhamnoides L. (Common name; Seabuckthorn), showed > 90% survival in mice population that was treated with lethal dose (10 Gy) of ⁶⁰Co gamma irradiation. In this study, early effects of pre-treatment with or without SBL-1 in blood peripheral blood lymphocytes (PBMCs) were investigated by cell viability assays (trypan blue and MTT). The quantitative in vitro study of Hoescht/PI staining was performed to check the apoptosis/necrosis in PBMCs irradiated at 2 Gy with or without pretreatment of SBL-1 (at different concentrations) up to 24 and 48h. Comet assay was performed in vivo, to detect the DNA strands breaks and its repair mechanism on peripheral blood lymphocytes at lethal dose (10 Gy). For this study, male mice (wt. 28 ± 2g) were administered radioprotective dose (30mg/kg body weight) of SBL-1, 30 min prior to irradiation. Animals were sacrificed at 24h and 48h. Blood was drawn through cardiac puncture, and blood lymphocytes were separated using histopaque column. Both neutral and alkaline comet assay were performed using standardized technique. In irradiated animals, alkaline comet assay revealed single strand breaks (SSBs) that showed significant (p < 0.05) increase in percent DNA in tail and Olive tail moment (OTM) at 24 h while at 48h the percent DNA in tail further increased significantly (p < 0.02). The double strands breaks (DSBs) increased significantly (p < 0.01) at 48 h in neutral assay, in comparison to untreated control. The animals pre-treated with SBL-1 before irradiation showed significantly (p < 0.05) less DSBs at 48 h treatment in comparison to irradiated group of animals. The SBL-1 alone treated group itself showed no toxicity. The antioxidant potential of SBL-1 were also investigated by in vitro biochemical assays such as DPPH (p < 0.05), ABTS, reducing ability (p < 0.09), hydroxyl radical scavenging (p < 0.05), ferric reducing antioxidant power (FRAP), superoxide radical scavenging activity (p < 0.05), hydrogen peroxide scavenging activity (p < 0.05) etc. SBL-1 showed strong free radical scavenging power that plays important role in the studies of radiation-induced injuries. The SBL-1 treated PBMCs showed significant (p < 0.02) viability in trypan blue assay at 24-hour incubation.

Keywords: radiation, SBL-1, SSBs, DSBs, FRAP, PBMCs

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Authors: Homayun Ghasemi, Mojtaba Yousefirad, Mozhgan Farzamisepehr

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Heavy metals are among soil pollutant resources that in case of accumulation in the soil and absorption by the plant, enter into the food chain and poison the plants or the people who consume those plants. This research was performed in order to examine the role of cadmium as a heavy metal in the activity of catalase and peroxidase as well as protein concentration in Trifolium resupinatum L. based on a randomized block design with three repetitions. The used treatments included consumption of Cd (NO3)2 at four levels, namely, 0, 100, 200, and 300 ppm. The plants under study were treated for 10 days. The results of the study showed that catalase activity decreased by the increase of cadmium. Moreover, peroxidase activity increased by an increase inthe consumption of cadmium. The analysis of protein level showed that plantlet protein decreased in high cadmium concentrations. The findings also demonstrated that cadmium concentration in roots was higher than in shoots.

Keywords: catalase, heavy metal, peroxidase, protein

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Authors: Asit K. Sikary, O. P. Murty

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Adipocere formation is a modification of the process of putrefaction. It consists mainly of saturated fatty acids, formed by the post-mortem hydrolysis and hydrogenation of body fats with the help of bacterial enzymes in the presence of warmth, moisture and anaerobic bacteria. In temperate climate, it takes weeks to develop while in India it starts to begin within 4-5 days. In this study, we have collected cases with adipocere formation, which were from the South Delhi region (average room temperature 27-390C) and autopsied at our centre. Details of the circumstances of the death, cause and time of death, surrounding environment and demographic profile of the deceased were taken into account. Total 16 cases were included in this study. Adipocere formation was predominantly present over cheeks, shoulder, breast, flanks, buttocks, and thighs. Out of 16, 11 cases were found in a dry atmosphere, 5 cases were brought from the water. There were 5 cases in which adipocere formation was seen in less than 2 days, and among them, in 1 case, as early as one day. This study showed that adipocere formation can be seen as early as 1 day in a hot and humid environment.

Keywords: adipocere, drowning, hanging, humid environment, strangulation, subtropical climate

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Authors: Azza El-Medany, Jamila El-Medany

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Residential and agricultural pesticide use is widespread in the world. Their extensive and indiscriminative use, in addition with their ability to interact with biological systems other than their primary targets constitute a health hazards to both humans and animals. The toxic effects of pesticides include alterations in metabolism; there is a lack of knowledge that organophosphates can cause pancreatic toxicity. The primary goal of this work is to study the effects of chronic exposure to Diazinon an organophosphate used in agriculture on pancreatic tissues and evaluate the ameliorating effect of Mesna as antioxidant on the toxicity of Diazinon on pancreatic tissues.40 adult male rats, their weight ranged between 300-350 g. The rats were classified into three groups; control (10 rats) was received corn oil at a dose of 1 0 mg/kg/day by gavage once a day for 2 months. Diazinon (15 rats) was received Diazinon at a dose of 10 mg/kg/day dissolved in corn oil by gavage once a day for 2 months. Treated group (15 rats), were received Mesna 180mg/kg once a week by gavage 15 minutes before administration of Diazinon for 2 months. At the end of the experiment, animals were anesthetized, blood samples were taken by cardiac puncture for glucose and insulin assays and pancreas was removed and divided into 3 portions; first portion for histopathological study; second portion for ultrastructural study; third portion for biochemical study using Elisa Kits including determination of malondialdehyde (MDA), tumor necrosis factor α (TNF-α), myeloperoxidase activity (MPO), interleukin 1β (IL-1β). A significant increase in the levels of MDA, TNF-α, MPO activity, IL-1β, serum glucose levels in the toxicated group with Diazinon were observed, while a significant reduction was noticed in GSH in serum insulin levels. After treatment with Mesna a significant reduction was observed in the previously mentioned parameters except that there was a significant rise in GSH in insulin levels. Histopathological and ultra-structural studies showed destruction in pancreatic tissues and β cells were the most affected cells among the injured islets as compared with the control group. The current study try to spot light about the effects of chronic exposure to pesticides on vital organs as pancreas also the role of oxidative stress that may be induced by them in evoking their toxicity. This study shows the role of antioxidant drugs in ameliorating or preventing the toxicity. This appears to be a promising approach that may be considered as a complementary treatment of pesticide toxicity.

Keywords: Diazinon, reduced glutathione, myeloperoxidase activity, tumor necrosis factor α, Mesna

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Authors: S. Ivanova, K. Ivanov

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Bemitil (2-ethylthiobenzimidazole hydrobromide) is a synthetic adaptogen and actoprotector, with wide-ranging pharmacological activities such as nootropic, antihypoxic, antioxidant, immunostimulant. The intake of Bemitil increases mental and physical performance and could be applied under either normal or extreme conditions. Until 2017 Bemitil was not considered as doping and was used by professional athletes more than 30 years because of its high efficiency and safety. The drug was included in WADA monitoring programme for 2018, and most likely it would be included in WADA Prohibited List for 2019. Usually, a substance/method is included in WADA Prohibited List if it meets any two of the following three criteria: the potential to enhance or enhances sports performance/ potential health risk to the athlete/ violates the spirit of sport. Bemitil has high performance-enhancing potential, but it is also safe- it is controversial whether it should be considered as doping.

Keywords: doping, bemitil, sport, actoprotector

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Authors: Andrés Quiroz, Emilio Hormazabal, Ana Mutis, Fernando Ortega, Manuel Chacón-Fuentes, Leonardo Parra

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Red clover (Trifolium pratense L.) is an important forage species in temperate regions of the world. The main limitation of this species worldwide is a lack of persistence related to the high mortality of plants due to a complex of biotic and abiotic factors, determining a life span of two or three seasons. Because of the importance of red clover in Chile, a red clover breeding program was started at INIA Carillanca Research Center in 1989, with the main objective of improving the survival of plants, forage yield, and persistence. The main selection criteria for selecting new varieties have been based on agronomical parameters and biotic factors. The main biotic factor associated with red clover mortality in Chile is Hylastinus obscurus (Coleoptera: Curculionidae). Both larval and adults feed on the roots, causing weakening and subsequent death of clover plants. Pesticides have not been successful for controlling infestations of this root borer. Therefore, alternative strategies for controlling this pest are a high priority for red clover producers. Currently, the role of semiochemical in the interaction between H. obscurus and red clover plants has been widely studied for our group. Specifically, from the red clover foliage has been identified limonene is eliciting repellency from the root borer. Limonene is generated in the plant from two independent biosynthetic pathways, the mevalonic acid, and deoxyxylulose pathway. Mevalonate pathway enzymes are localized in the cytosol, whereas the deoxyxylulose phosphate pathway enzymes are found in plastids. In summary, limonene can be determinated by enzymatic bioassay using GPP as substrate and by limonene synthase expression. Therefore, the main objective of this work was to study genetic variation of limonene in material provided by INIA´s Red Clover breeding program. Protein extraction was carried out homogenizing 250 mg of leave tissue and suspended in 6 mL of extraction buffer (PEG 1500, PVP-30, 20 mM MgCl2 and antioxidants) and stirred on ice for 20 min. After centrifugation, aliquots of 2.5 mL were desalted on PD-10 columns, resulting in a final volume of 3.5 mL. Protein determination was performed according to Bradford with BSA as a standard. Monoterpene synthase assays were performed with 50 µL of protein extracts transferred into gas-tight 2 mL crimp seal vials after addition of 4 µL MgCl₂ and 41 µL assay buffer. The assay was started by adding 5 µL of a GPP solution. The mixture was incubated for 30 min at 40 °C. Biosynthesized limonene was quantified in a GC equipped with a chiral column and using synthetic R and S-limonene standards. The enzymatic the production of R and S-limonene from different Superqueli-Carillanca genotypes is shown in this work. Preliminary results showed significant differences in limonene content among the genotypes analyzed. These results constitute an important base for selecting genotypes with a high content of this repellent monoterpene towards H. obscurus.

Keywords: head space, limonene enzymatic determination, red clover, Hylastinus obscurus

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Authors: I. Tulay Cagatay

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Blue-green microalgae cyanobacteria, which are important photosynthetic organisms of aquatic ecosystems, are the primary sources of many bioactive compounds such as proteins, carbohydrates, lipids, vitamins and enzymes that can be used as antimicrobial and antiviral agents. Some of these organisms are nowadays used directly in the food, cosmetic and pharmaceutical industry, or in aquaculture and biotechnological approaches like biofuel or drug therapy. Finding the effective, environmental friendly chemotropic and antimicrobial agents to control fish pathogens are crucial in a country like Turkey which has a production capacity of about 240 thousand tons of cultured fish and has 2377 production farms and which is the second biggest producer in Europe. In our study, we tested the antimicrobial activity of cyanobacterium spp. against some fish pathogens Aeromonas hydrophila and Yersinia ruckeri that are important pathogens for rainbow trout farms. Agar disk diffusion test method was used for studying antimicrobial activity on pathogens. Both tested microorganisms have shown antimicrobial activity positively as the inhibition zones were 0.45 mm and 0.40 mm respectively.

Keywords: fish pathogen, cyanobacteria, antimicrobial activity, trout

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Authors: Niharika Kaushal, Minni Singh

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Citrus fruits contain an abundance of phytochemicals that can promote health. A substantial amount of agrowaste is produced from the juice processing industries, primarily peels and seeds. This leftover agrowaste is a reservoir of nutraceuticals, particularly bioflavonoids which render it antioxidant and potentially anticancerous. It is, therefore, favorable to utilize this biomass and contribute towards sustainability in a manner that value-added products may be derived from them, nutraceuticals, in this study. However, the pre-systemic metabolism of flavonoids in the gastric phase limits the effectiveness of these bioflavonoids derived from mandarin biomass. In this study, ‘kinnow’ mandarin (Citrus nobilis X Citrus deliciosa) biomass was explored for its flavonoid profile. This work entails supercritical fluid extraction and identification of bioflavonoids from mandarin biomass. Furthermore, to overcome the limitations of these flavonoids in the gastrointestinal tract, a double-layered vehicular mechanism comprising the fabrication of nanoconjugates and edible hydrogels was adopted. Total flavonoids in the mandarin peel extract were estimated by the aluminum chloride complexation method and were found to be 47.3±1.06 mg/ml rutin equivalents as total flavonoids. Mass spectral analysis revealed the abundance of polymethoxyflavones (PMFs), nobiletin and tangeretin as the major flavonoids in the extract, followed by hesperetin and naringenin. Furthermore, the antioxidant potential was analyzed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method, which showed an IC50 of 0.55μg/ml. Nanoconjugates were fabricated via the solvent evaporation method, which was further impregnated into hydrogels. Additionally, the release characteristics of nanoconjugate-laden hydrogels in a simulated gastrointestinal environment were studied. The PLGA-PMFs nanoconjugates exhibited a particle size between 200-250nm having a smooth and spherical shape as revealed by FE-SEM. The impregnated alginate hydrogels offered a dense network that ensured the holding of PLGA-PMF nanoconjugates, as confirmed by Cryo-SEM images. Rheological studies revealed the shear-thinning behavior of hydrogels and their high resistance to deformation. Gastrointestinal studies showed a negligible 4.0% release of flavonoids in the gastric phase, followed by a sustained release over the next hours in the intestinal environment. Therefore, based on the enormous potential of recovering nutraceuticals from agro-processing wastes, further augmented by nanotechnological interventions for enhancing the bioefficacy of these compounds, lays the foundation for exploring the path towards the development of value-added products, thereby contributing towards the sustainable use of agrowaste.

Keywords: agrowaste, gastrointestinal, hydrogel, nutraceuticals

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Authors: Han-Bin Kim, Sooim Shin, Moonsung Choi

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There is a growing interest in introducing a desired functional group on enzymes in the field of protein engineering. In here, various redox centers were newly created using histidine tag, which is widely used for protein purification, plus cobalt in one of cupredoxins, amicyanin. The coordination of Cobalt-His tag and reactivity of the Co²⁺ loaded His-tag also were characterized. 3xHis-tag, 6xHis-tag, and 9xHis-tag were introduced on amicyanin by site-directed mutagenesis, and then Co²⁺ was loaded on each His-tagged amicyanin. The spectral changes at 330 nm corresponding to cobalt binding on His-tag site indicated the binding ratio of 3xHis-tag, 6xHis-tag, and 9xHis-tag to cobalt as 1:1, 1:2, 1:3 respectively. Based on kinetic studies of binding cobalt to 3xHis-tag, 6xHis-tag, and 9xHis-tagged amicyanin, the nature of the sites was elucidated. In addition, internal electron transfer properties between Cu¹⁺ site and engineered site of amicyanin were determined. These results provide insight into improvement of metal coordination and alternation of the redox properties of metal as a new catalytic site on proteins.

Keywords: amicyanin, cobalt, histidine, protein engineering

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Authors: Serebryanaya Fatima, Essaih Hind

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The Rosa damascena (Rosa damascena Mill.) is an interesting medicinal plant; it is famous in different countries and has medicinal use in many cultures. The main groups of pharmacological actions of rose oil are connected with anti-inflammatory, antifungal activity, also antioxidant and antibacterial, and antiparasitic properties. We have prepared the quality indicators analysis of the Damascus rose oil. An iodine number, acid number, and oil peroxide number were determined. The following indicators of the quality of rose oil have been studied. The determination was carried out according to the pharmacopoeic methods of analysis of essential oils, the definition of peroxide number (1,971%), iodine number (3,365%), and acid number (0,0526%).

Keywords: Rosa damascene, Rosa damascena Mill., iodine number, acid number, oil peroxide number

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