Search results for: anti-oxidant metabolism

Authors: Paresh Patel, Harshit Patel

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Superficial fungal skin infection is among the most common skin disease. The drug administration through skin has received attention due to several advantages: Avoidance of significant pre-systemic metabolism, drug levels within the therapeutic window, drugs with short biological half-lives, decreased side effects, the non-invasive character, and very high acceptance.

Keywords: transdermal spray, ketoconazole, Eudragit® RLPO, therapeutic window

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Authors: Anupama Sahoo, Bongyong Lee, Katia Boniface, Julien Seneschal, Sanjaya K. Sahoo, Tatsuya Seki, Chunyan Wang, Soumen Das, Xianlin Han, Michael Steppie, Sudipta Seal, Alain Taieb, Ranjan J. Perera

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Vitiligo is a common, chronic skin disorder characterized by loss of epidermal melanocytes and progressive depigmentation. Vitiligo has a complex immune, genetic, environmental, and biochemical etiology, but the exact molecular mechanisms of vitiligo development and progression, particularly those related to metabolic control, are poorly understood. Here we characterized the human vitiligo cell line PIG3V and the normal human melanocytes, HEM-l by RNA-sequencing, targeted metabolomics, and shotgun lipidomics. Melanocyte-enriched miR-211, a known metabolic switch in non-pigmented melanoma cells, was severely downregulated in vitiligo cell line PIG3V and skin biopsies from vitiligo patients, while its novel predicted targets transcriptional co-activator PGC1-α (PPARGC1A), ribonucleotide reductase regulatory subunit M2 (RRM2), and serine-threonine protein kinase TAO1 (TAOK1) were reciprocally upregulated. miR-211 binds to PGC1-α 3’UTR locus and represses it. Although mitochondrial numbers were constant, mitochondrial complexes I, II, and IV and respiratory responses were defective in vitiligo cells. Nanoparticle-coated miR-211 partially augmented the oxygen consumption rate in PIG3V cells. The lower oxygen consumption rate, changes in lipid and metabolite profiles, and increased reactive oxygen species production observed in vitiligo cells appear to be partly due to abnormal regulation of miR-211 and its target genes. These genes represent potential biomarkers and therapeutic targets in human vitiligo.

Keywords: metabolism, microRNA, mitochondria, vitiligo

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Authors: D. M. El-Tanbouly, R. M. Abdelsalam, A. S. Attia, M. T. Abdel-Aziz

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Lipopolysaccharide (LPS) endotoxin, a component of the outer membrane of Gram-negative bacteria, is involved in the pathogenesis of sepsis. LPS administration induces systemic inflammation that mimics many of the initial clinical features of sepsis and has deleterious effects on several organs including the liver and eventually leading to septic shock and death. The present study aimed to investigate the protective effect of magnesium, a well-known cofactor in many enzymatic reactions and a critical component of the antioxidant system, on hepatic damage associated with LPS induced- endotoxima in mice. Mg (20 and 40 mg/kg, po) was administered for 7 consecutive days. Systemic inflammation was induced one hour after the last dose of Mg by a single dose of LPS (2 mg/kg, ip) and three hours thereafter plasma was separated, animals were sacrificed and their livers were isolated. LPS-treated mice suffered from hepatic dysfunction revealed by histological observation, elevation in plasma transaminases activities, C-reactive protein content and caspase-3, a critical marker of apoptosis. Liver inflammation was evident by elevation in liver cytokines contents (TNF-α and IL-10) and myeloperoxidase (MPO) activity. Additionally, oxidative stress was manifested by increased liver lipoperoxidation, glutathione depletion, elevated total nitrate/nitrite (NOx) content and glutathione peroxidase (GPx) activity. Pretreatment with Mg largely mitigated these alternations through its anti-inflammatory and antioxidant potentials. Mg, therefore, could be regarded as an effective strategy for prevention of liver damage associated with septicemia.

Keywords: LPS, liver damage, magnesium, septicemia

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Authors: Hanan A. Soliman, Mohamed A. El-Desouky, Walaa G. Hozayen, Rasha R. Ahmed, Amal K . Khaliefa

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Aim: The objective of this study is to investigate the hypoglycemic, hypolipidemic, and hepatoprotective effects of the aqueous extract of parsley, basil, and chicory whole plant in normal and dexamethasone (Dex) rats. Materials and Methods: 50 female albino rats were used in this study and divided into 5 groups (for each 10). Group (1) fed basal diet and maintained as negative control group. Group (2) received Dex in a dose of (0.1 mg/kg b. wt.). Groups 3, 4, and 5 were treated with Dex along with three different plant extracts of parsley, basil, and chicory (2 g/kg b. wt.), (400 mg/kg b. wt.), and (100 mg/kg b. wt.), respectively. Results: All these groups were treated given three times per week for 8 consecutive weeks. Dex-induced alterations in the levels of serum glucose, triglyceride, cholesterol, low-density lipoprotein-cholesterol levels and cardiovascular indices and serum alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase activities, liver thiobarbituric acid (TBARS) levels increased, while high-density lipoprotein-cholesterol, total protein, albumin, and liver glutathione (GSH) levels decreased. On the other hand, plant extracts succeeded to modulate these observed abnormalities resulting from Dex as indicated by the reduction of glucose, cholesterol, TBARS, and the pronounced improvement of the investigated biochemical and antioxidant parameters. Conclusions: It was concluded that probably, due to its antioxidant property, parsley, basil, and chicory extracts have hepatoprotective effects in Dex-induced in rats.

Keywords: antioxidants, dexamethasone, hyperglycemia, hyperlipidemia

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Authors: Sri Rejeki, Tamrin Tamrin, RH. F. Faradilla, Muhammad N. Ibrahim, Mariana M., Irnawati Irnawati

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Cocoa bean shells, despite their antioxidant and antimicrobial properties, are still considered as an underutilized agricultural waste. The functional properties and their lignocelluloses content make cocoa bean shells a potential material for paper-based food packaging. In our previous research, we have successfully produced papers from cocoa bean shells that had antioxidant and antibacterial activities. However, the hydrophilic nature of the lignocelluloses of cocoa bean shells hinders the application of the paper to be used as a food packaging. In this research, we aimed to study the effects of beeswax coating on the wettability and mechanical properties of the paper. The coating was done by dipping the papers in beeswax solution several times and in three different beeswax concentrations. The number of dipping and beeswax concentration significantly (p<0.05) affected the water contact angle of the papers. Results show that the water contact angle increases dramatically due to the coating treatment. The control paper or uncoated paper had a contact angle of 40.50o, while the contact angle of the best-coated paper (D3B3: 3x dipping, 3g/10mL beeswax) reached 96.93o. Both tensile strength and percent elongation were not significantly (p>0.05) affected by the coating treatment. This showed that beeswax was a potential organic material to improve the hydrophobicity of paper from cocoa bean shells without any undesirable effects on the mechanical properties of the paper.

Keywords: cocoa bean shell, paper, beeswax, coating, contact angle

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Authors: Aneta Maćczak, Bożena Bukowska, Jaromir Michałowicz

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Bisphenols are one of the most widely used chemical compounds worldwide. They are used in the manufacturing of polycarbonates, epoxy resins and thermal paper which are applied in plastic containers, bottles, cans, newspapers, receipt and other products. Among these compounds, bisphenol A (BPA) is produced in the highest amounts. There are concerns about endocrine impact of BPA and its other toxic effects including hepatotoxicity, neurotoxicity and carcinogenicity on human organism. Moreover, BPA is supposed to increase the incidence the obesity, diabetes and heart disease. For this reason the use of BPA in the production of plastic infant feeding bottles and some other consumers products has been restricted in the European Union and the United States. Nowadays, BPA analogues like bisphenol F (BPF) and bisphenol S (BPS) have been developed as alternative compounds. The replacement of BPA with other bisphenols contributed to the increase of the exposure of human population to these substances. Toxicological studies have mainly focused on BPA. In opposite, a small number of studies concerning toxic effects of BPA analogues have been realized, which makes impossible to state whether those substituents are safe for human health. Up to now, the mechanism of bisphenols action on the erythrocytes has not been elucidated. That is why, the aim of this study was to assess the effect of BPA and its selected analogues such as BPF and BPS on the activity of antioxidant enzymes, i.e. catalase (EC 1.11.1.6.), glutathione peroxidase (E.C.1.11.1.9) and superoxide dismutase (EC.1.15.1.1) in human erythrocytes. Red blood cells in respect to their function (transport of oxygen) and very well developed enzymatic and non-enzymatic antioxidative system, are useful cellular model to assess changes in redox balance. Erythrocytes were incubated with BPA, BPF and BPS in the concentration ranging from 0.5 to 100 µg/ml for 24 h. The activity of catalase was determined by the method of Aebi (1984). The activity of glutathione peroxidase was measured according to the method described by Rice-Evans et al. (1991), while the activity of superoxide dismutase (EC.1.15.1.1) was determined by the method of Misra and Fridovich (1972). The results showed that BPA and BPF caused changes in the antioxidative enzymes activities. BPA decreased the activity of examined enzymes in the concentration of 100 µg/ml. We also noted that BPF decreased the activity of catalase (5-100 µg/ml), glutathione peroxidase (50-100 µg/ml) and superoxide dismutase (25-100 µg/ml), while BPS did not cause statistically significant changes in investigated parameters. The obtained results suggest that BPA and BPF disrupt redox balance in human erythrocytes but the observed changes may occur in human organism only during occupational or subacute exposure to these substances.

Keywords: antioxidant enzymes, bisphenol A, bisphenol a analogues, human erythrocytes

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Authors: Laura M. Hovsepyan, Gayane S. Ghazaryan, Hasmik V. Zanginyan

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Hypertension (HD) is the most common cardiovascular pathology that causes disability and mortality in the working population. Most often, heart failure (HF), which is based on myocardial remodeling, leads to death in hypertension. Recently, endothelial dysfunction (EDF) or a violation of the functional state of the vascular endothelium has been assigned a significant role in the structural changes in the myocardium and the occurrence of heart failure in patients with hypertension. It has now been established that tissues affected by inflammation form increased amounts of superoxide radical and NO, which play a significant role in the development and pathogenesis of various pathologies. They mediate inflammation, modify proteins and damage nucleic acids. The aim of this work was to study the processes of oxidative modification of proteins (OMP) and the production of nitric oxide in hypertension. In the experimental work, the blood of 30 donors and 33 patients with hypertension was used. For the quantitative determination of OMP products, the based on the reaction of the interaction of oxidized amino acid residues of proteins and 2,4-dinitrophenylhydrazine (DNPH) with the formation of 2,4-dinitrophenylhydrazones, the amount of which was determined spectrophotometrically. The optical density of the formed carbonyl derivatives of dinitrophenylhydrazones was recorded at different wavelengths: 356 nm - aliphatic ketone dinitrophenylhydrazones (KDNPH) of neutral character; 370 nm - aliphatic aldehyde dinirophenylhydrazones (ADNPH) of neutral character; 430 nm - aliphatic KDNFG of the main character; 530 nm - basic aliphatic ADNPH. Nitric oxide was determined by photometry using Grace's solution. Adsorption was measured on a Thermo Scientific Evolution 201 SF at a wavelength of 546 nm. Thus, the results of the studies showed that in patients with arterial hypertension, an increased level of nitric oxide in the blood serum is observed, and there is also a tendency to an increase in the intensity of oxidative modification of proteins at a wavelength of 270 nm and 363 nm, which indicates a statistically significant increase in aliphatic aldehyde and ketone dinitrophenylhydrazones. The increase in the intensity of oxidative modification of blood plasma proteins in the studied patients, revealed by us, actually reflects the general direction of free radical processes and, in particular, the oxidation of proteins throughout the body. A decrease in the activity of the antioxidant system also leads to a violation of protein metabolism. The most important consequence of the oxidative modification of proteins is the inactivation of enzymes.

Keywords: hypertension (HD), oxidative modification of proteins (OMP), nitric oxide (NO), oxidative stress

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Authors: Tamer El-Messery, Teresa Sanchez-Moya, Ruben Lopez-Nicolas, Gaspar Ros, Esmat Aly

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Olive leaves (OLs), the main by-product of the olive oil industry, have a considerable amount of phenolic compounds. The exploitation of these compounds represents the current trend in food processing. In this study, OLs polyphenols were microencapsulated with polycaprolactone (PCL) and utilized in formulating novel functional yoghurt. PCL-microcapsules were characterized by scanning electron microscopy, and Fourier transform infrared spectrometry analysis. Their total phenolic (TPC), total flavonoid (TFC) contents, and antioxidant activities (DPPH, FRAP, ABTS), and polyphenols bioaccessibility were measured after oral, gastric, and intestinal steps of in vitro digestion. The four yoghurt formulations (containing 0, 25, 50, and 75 mg of PCL-microsphere/100g yoghurt) were evaluated for their pH, acidity, syneresis viscosity, and color during storage. In vitro digestion significantly affected the phenolic composition in non-encapsulated extract while had a lower impact on encapsulated phenolics. Higher protection was provided for encapsulated OLs extract, and their higher release was observed at the intestinal phase. Yoghurt with PCL-microsphere had lower viscosity, syneresis, and color parameters, as compared to control yoghurt. Thus, OLs represent a valuable and cheap source of polyphenols which can be successfully applied, in microencapsulated form, to formulate functional yoghurt.

Keywords: yoghurt quality attributes, olive leaves, phenolic and flavonoids compounds, antioxidant activity, polycaprolactone as microencapsulant

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Authors: Marwa T. Badawy, Alaa F. Bakr, Nesrine Hegazi, Mohamed A. Farag, Ahmed Abdellatif

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Diabetes Mellitus (DM) is a chronic disease affecting a large population worldwide. Africa is rich in native medicinal plants with myriad health benefits, though less explored towards the development of specific drug therapy as in diabetes. This study aims to determine the in vivo antidiabetic potential of the well-reported and traditionally used fruits of Baobab (Adansonia digitata L.) using STZ induced diabetic model. The in-vitro cytotoxic and antioxidant properties were examined using MTT assay on L-929 fibroblast cells and DPPH antioxidant assays, respectively. The extract showed minimal cytotoxicity with an IC50 value of 105.7 µg/mL. Histopathological and immunohistochemical investigations showed the hepatoprotective and the renoprotective effects of A. digitata fruits’ extract, implying its protective effects against diabetes complications. These findings were further supported by biochemical assays, which showed that i.p., injection of a low dose (150 mg/kg) of A. digitata twice a week lowered the fasting blood glucose levels, lipid profile, hepatic and renal markers. For a comprehensive overview of extract metabolites composition, ultrahigh performance (UHPLC) analysis coupled to high-resolution tandem mass spectrometry (HRMS/MS) in synchronization with molecular networks led to the annotation of 77 metabolites, among which 50% are reported for the first time in A. digitata fruits.

Keywords: adansonia digital, diabetes mellitus, metabolomics, streptozotocin, Sprague, dawley rats

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Authors: Gianella Franco, Karen Suarez, María Quijano, Patricia Manzano

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Agro-industries generate large amounts of waste, which are mostly untapped. This research was carried out to use cocoa, coffee and orange industrial by-products to develop a dairy drink. The product was prepared by making a 10% aqueous extract of the mixture of cocoa and coffee beans shells and orange peel. Extreme Vertices Mixture Design was applied to vary the proportions of the ingredients of the aqueous extract, getting 13 formulations. Each formulation was mixed with skim milk and pasteurized. The attributes of taste, smell, color and appearance were evaluated by a semi-trained panel by multiple comparisons test, comparing the formulations against a standard marked as "R", which consisted of a coffee commercial drink. The formulations with the highest scores were selected to maximize the Total Polyphenol Content (TPC) through a process of linear optimization resulting in the formulation 80.5%: 18.37%: 1.13% of cocoa bean shell, coffee bean shell and orange peel, respectively. The Total Polyphenol Content was 4.99 ± 0.34 mg GAE/g of drink, DPPH radical scavenging activity (%) was 80.14 ± 0.05 and caffeine concentration of 114.78 mg / L, while the coffee commercial drink presented 3.93 ± 0.84 mg GAE / g drink, 55.54 ± 0.03 % and 47.44 mg / L of TPC, DPPH radical scavenging activity and caffeine content, respectively. The results show that it is possible to prepare an antioxidant - rich drink with good sensorial attributes made of industrial by-products.

Keywords: DPPH, polyphenols, waste, food science

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Authors: Harshada Vaidya-Kannur, Dattatraya Naik

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Propolis, one of the beehive products also referred as bee-glue is sticky dark coloured complex mixture of compounds. The volatile oil can be isolated from the propolis by hydrodistillation. The mark that is left behind after the removal of volatile oil is referred as the de-oiled propolis. Antioxidant as well as anti-inflammatory properties of total ethanolic extract of de-oiled propolis (TEEDP) was investigated. Another lot of deoiled propolis was successively exacted with hexane, ethyl acetate and ethanol. Activities of these fractions were also determined. Antioxidant activity was determined by studying ABTS, DPPH and NO radical scavenging. Determination of anti-inflammatory activity was carried out by topical TPA induced mouse ear oedema model. It is noteworthy that ethyl acetate fraction of deoiled propolis (EAFDP) exhibited 49.45 % TEAC activity at the concentration 0.2 mg/ml which is equivalent to the activity of trolox at the concentration 0.2 mg/ml. Its DPPH scavenging activity (72.56%) was closely comparable to that of trolox (75%). However its NO scavenging activity was comparatively low. From IC50 values it could be concluded that the efficiency of scavenging ABTS radicals by the de-oiled propolis was more pronounced as compared to scavenging of other radicals. Studies by TPA induced mouse ear inflammation model indicated that the de-oiled propolis of Indian origin had significant topical anti-inflammatory activity. The EAFDP was found to be the most active fraction for this activity also. The purification of EAFP yielded six pure crystalline compounds. These compounds were identified by their physical data and spectral data.

Keywords: anti-inflammatory activity, anti-oxidant activity, column chromatography, de-oiled propolis

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Authors: Fadime Eryılmaz Pehlivan

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Anthocyanins are important plant pigments having roles in many physiological and ecological functions; that are controlled by numerous regulatory factors. The accumulation of anthocyanins in Z. mays cause the plants stems to exhibit red coloration when encountering gradually increasing copper treatments (1, 5, and 10 mM of Cu in a period of 5 days) on maize seedlings. Stress injury was measured in terms of chlorophyll (a and b), carotenoid and anthocyanin contents, malondialdehyde (MDA), hydrogen peroxide (H2O2). Carotenoid and anthocyanin contents dramatically increased by increasing concentrations of Cu stress. MDA and H2O2 levels were found to significantly increase at high Cu treatments (5 and 10 mM of Cu). Chlorophyll content was observed to be highest at 1 mM Cu and then decreased at 5 and 10 mM of Cu. In addition, significant increases were determined in the activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and ascorbate peroxidase (APX) under high Cu concentrations, while glutathione S-transferase (GST) and peroxidase (POX) activities showed no change. Treatments above 5 and 10 mM of Cu triggered copper stress in maize seedlings. The results of this study provide evidence that maize seedlings represent a high tolerance to gradually increasing copper treatments. Improved copper tolerance may relate to high anthocyanin, and carotenoid content besides antioxidant enzyme activity may improve the metal chelating ability of anthocyanin pigments. Data presented in this study may also contribute to a better understanding of phytoremediation studies in maize exposed to high copper contenting soils.

Keywords: anthocyanin, copper, maize , antioxidant

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Authors: Surendra Kumar Gautam, Mahesh Dhungana

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Magnesium oxide nanoparticles (MgO NPs) are less toxic to humans and the environment as compared to other metal oxide nanoparticles. Various conventional chemical and physical methods are used for synthesis whose toxicity level is high and highly expensive. As the best alternative, phyto-assisted synthesis has emerged, which uses extracts from plant parts for the synthesis of nanoparticles. Here, we report the synthesis of MgO nanoparticles with the assistance of beetroot extract and leaf extract of P. guajava and A. adenophora. The synthesized MgO NPs were characterized by X-ray diffraction (XRD), Fourier transforms infrared spectroscopy (FTIR), and UV-visible spectroscopy. X-ray analysis for the broadening of peaks was used to evaluate the crystallite size and lattice strain using Debye-Scherer and Williamson–Hall method. The results of crystallite size obtained by both methods are in close proximity. The crystallite size obtained by the Williamson-Hall method seems more accurate, with values being 8.1 nm and 13.2 nm for beetroot MgO NPs and P. guajava MgO NPs, respectively. The FT-IR spectroscopy revealed the dominance of chemical bonds as well as functional groups on MgO NPs surfaces. The UV-visible absorption spectra of MgO NPs were found to be 310 nm, 315 nm, and 315 nm for beetroot, P. guajava, and A. adenophora leaf extract, respectively. Among the three samples, beetroot-mediated MgO NPs were effective antibacterial against both gram-positive and Gram-negative bacteria. In addition, synthesized MgO NPs also show significant antioxidant efficacy against 1,1-diphenyl-2-picrylhydrazyl radical. Further, beetroot MgO NPs showed the highest photocatalytic activity of about 91% in comparison with other samples.

Keywords: MgO NPs, XRD, FTIR, antibacterial, antioxidant and photocatalytic activity

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Authors: Tatyane Lopes de Freitas, Antonio Diogo S. Vieira, Susana Marta Isay Saad, Maria Ines Genovese

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The fruit processing industries generate a large volume of residues to produce juices, pulps, and jams. These residues, or by-products, consisting of peels, seeds, and pulps, are routinely discarded. Fruits are rich in bioactive compounds, including polyphenols, which have positive effects on health. Dry residues from two fruits, acerola (M. emarginata D. C.) and orange (C. sinensis), were characterized in relation to contents of ascorbic acid, minerals, total dietary fibers, moisture, ash, lipids, proteins, and carbohydrates, and also high performance liquid chromatographic profile of flavonoids, total polyphenols and proanthocyanidins contents, and antioxidant capacity by three different methods (Ferric reducing antioxidant power assay-FRAP, Oxygen Radical Absorbance Capacity-ORAC, 1,1-diphenyl-2-picrylhydrazil (DPPH) radical scavenging activity). Acerola by-products presented the highest acid ascorbic content (605 mg/100 g), and better antioxidant capacity than orange by-products. The dry residues from acerola demonstrated high contents of proanthocyanidins (617 µg CE/g) and total polyphenols (2525 mg gallic acid equivalents - GAE/100 g). Both presented high total dietary fiber (above 60%) and protein contents (acerola: 10.4%; orange: 9.9%), and reduced fat content (acerola: 1.6%; orange: 2.6%). Both residues showed high levels of potassium, calcium, and magnesium, and were considered sources of these minerals. With acerola by-product, four formulations of probiotics fermented milks were produced: F0 (without the addition of acerola residue (AR)), F2 (2% AR), F5 (5% AR) and F10 (10% AR). The physicochemical characteristics of the fermented milks throughout of storage were investigated, as well as the impact of in vitro simulated gastrointestinal conditions on flavonoids and probiotics. The microorganisms analyzed maintained their populations around 8 log CFU/g during storage. After the gastric phase of the simulated digestion, the populations decreased, and after the enteric phase, no colonies were detected. On the other hand, the flavonoids increased after the gastric phase, maintaining or suffering small decrease after enteric phase. Acerola by-products powder is a valuable ingredient to be used in functional foods because is rich in vitamin C, fibers and flavonoids. These flavonoids appear to be highly resistant to the acids and salts of digestion.

Keywords: acerola, orange, by-products, fermented milk

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Authors: Qinghua Xing, Xinyi Tao, Haisheng Wang, Baisuo Zhao

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The first true anaerobic, halophilic alkali thermophile, Natranaerobius thermophilus DSM 18059T, serves as a valuable model for studying cellular adaptations to saline, alkaline and thermal extremes. To uncover the adaptive strategies employed by N. thermophilus in coping with these challenges, we conducted a comprehensive iTRAQ-based quantitative proteomic analysis under different conditions of salinity (3.5 M vs. 2.5 M Na+), pH (pH 9.6 vs. pH 8.6), and temperature (52°C vs. 42°C). The increased intracellular accumulation of glycine betaine, through both synthesis and transport, plays a critical role in N. thermophilus' adaptation to these combined stresses. Under all three stress conditions, the up-regulation of Trk family proteins responsible for K+ transport is observed. Intracellular K+ concentration rises in response to salt and pH levels. Multiple types of Na+/H+ antiporter (NhaC family, Mrp family and CPA family) and a diverse range of FOF1-ATP synthase are identified as vital components for maintaining ionic balance under different stress conditions. Importantly, proteins involved in amino acid metabolism, carbohydrate metabolism, ABC transporters, signaling and chemotaxis, as well as biological macromolecule repair and protection, exhibited significant up-regulation in response to these extreme conditions. These metabolic pathways emerge as critical factors in N. thermophilus' adaptation mechanisms under extreme environmental stress. To validate the proteomic data, ddPCR analysis confirmed changes in mRNA expression, thereby corroborating the up-regulation and down-regulation patterns of 19 co-up-regulated and 36 key proteins under saline, alkaline and thermal stresses. This research enriches our understanding of the complex regulatory systems that enable polyextremophiles to survive in combined extreme conditions.

Keywords: polyextremophiles, natranaerobius thermophilus, saline- alkaline- thermal stresses, combined extremes

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Authors: Lalit Kishore, Randhir Singh

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Diabetic nephropathy (DN) is the major cause of morbidity among diabetic patients. In this study, the effect of Bacopa monnieri Linn. (Brahmi, BM), was studied in a Streptozotocin (STZ)-induced experimental rat model of DN. Diabetic nephropathy was induced in Male Wistar rats (body weight- 300± 10 gms) by single intra-peritoneal injection of STZ (45mg/kg, i.p.) after 15 min of Nicotinamide (230 mg/kg) administration. Different doses of alcoholic extract i.e. 100, 200 and 400 mg/kg was given for 45 days by oral gavage after induction of DN. Blood glucose level, serum insulin, glycosylated haemoglobin, renal parameters (serum urea, uric acid, creatinine and BUN) and lipid profile (total cholesterol, triglycerides, HDL, LDL and VLDL levels) were measured. Concentration of thiobarbituric acid reactive species (TBARS) and levels of antioxidant enzymes of reduced glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) were evaluated in the kidney, liver and pancreas. At the end of treatment period the alcoholic extract of BM reduced the elevated level of blood glucose, serum insulin, renal parameters, lipid levels, TBARS, AGE’s in kidney and significantly increased body weight, HDL and antioxidant enzymes in dose dependent manner as compared to diabetic control animals. These results suggested the BM possesses significant renoprotective activity.

Keywords: AGE's, lipid profile, oxidative stress, renal parameters

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Authors: Sadia Roshan, Kulsoom Sughra, Shazia Shamas, Shamaila Irum, Haleema Sadia

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To evaluate synergistic medicinal effects of Withania somnifera (Ashwaghandha) and Murraya koenigii (Curry pata) in vitro. Antimicrobial activity was determined using the disc diffusion method against five bacterial and two fungal strains. The antioxidant activity was evaluated by the DPPH assay. The antidiabetic activity was evaluated by alpha-glucosidase inhibition assay and alpha-amylase inhibition assay. Synergistic antibacterial activity was observed against all the strains of bacteria, either Gram-positive or Gram-negative and fungi under study conditions. The maximum antibacterial activity was displayed by combined extract against E. coli i.e. 26±0.4mm. Maximum antifungal activity was shown by combined extract against Aspergillus niger, i.e., 17.3±0.5mm. The antioxidant activity of the combined extract was also significant. Alpha-glucosidase inhibition and alpha-amylase inhibition assays also showed synergism. Results indicate that Withania somnifera and Murraya koengii have medicinal properties. The combined extract of both plants is more potent than their individual extracts, suggesting that these can work in synergism. The research suggests that different plant extracts could be used in combination to increase their medicinal activities by many folds, thus giving an insight into future use of herbal medication.

Keywords: withania somnifera, murraya koenigii, antimicrobial activity, gram-positive bacetria, gram-negative bacteria

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Authors: O. M. M. Sabry, Abeer M. El Sayed

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The essential oils from the aerial parts and flowers of Haplophyllum tuberculatum (Forsskal) Adr. Juss (Rutaceae) growing in Libya were obtained separately by hydro-distillation using a Clevenger-type apparatus. The essential oils yield were (0.4, 1.5w/w%) respectively based on the dry weight of the plant. The oils were analyzed by GC-MS. Twenty four constituents, amounting to 96.6%, were identified in the oil of the aerial parts. The predominant compounds were among the non oxygenated terpenoids (82.4%) as monoterpene hydrocarbons, represented by sabinen (26.4 %), δ-terpinen (26 %), β-phellandrene (10.4%) and 3-carene (3.86%). Zingiberine (0.4%) and β-sesquiphellandrene (0.12%) were the major sesquiterpene hydrocarbons identified. Oxygenated monoterpenes were represented by eucalyptol (5.5%) and piperitone (5.55%). Twenty six constituents, equivalent to 99.5%, were identified in the oil of the flowers. The dominance of monoterpene hydrocarbons in the flowers oil can be attributed to the high percentage of γ-terpinen (38.44%), β-phellandrene (10.0%), α- phellandrene (2.33%), 3,4-dimethyl-1,5-cyclooctadiene (6.67%), β-myrecene (6.04%), 3-carene (5.43%) and α-pinene (1.3%).While the oxygenated monoterpenes can be contributed to the trans-piperitol (4.67%) and piperitone (2.07%). Sesquiterpene hydrocarbons were not identified in the oil of the flower of H. tuberculatum. Variation in constitution between oils of Libyan H. tuberculatum and that obtained from other countries can be due to both environmental and genetic factors. The essential oils have demonstrated variable antimicrobial activities against certain micro-organisms. Also have revealed marked in vitro cytotoxicity against lung (H1299), liver (HEPG2) carcinoma cell line and variably effective as anti-inflammatory and antioxidant.

Keywords: Halpophyllum tuberculatum, rutaceae, essential oil, antimicrobial, anti-inflammatory, antitumor, antioxidant, Libya

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Authors: Eva Karaskova, Jana Volejnikova, Dusan Holub, Maria Velganova-Veghova, Michaela Spenerova, Dagmar Pospisilova

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Background: Hepcidin is the central regulator of iron metabolism. Its production is mainly affected by an iron deficiency and the presence of inflammatory activity in the body. The aim of this study was to compare serum hepcidin levels in paediatric patients with newly diagnosed inflammatory bowel disease and hepcidin levels during maintenance therapy, correlate changes of serum hepcidin levels with selected markers of iron metabolism and inflammation and type of provided treatment. Methods: Children with newly diagnosed Crohn's disease (CD) and ulcerative colitis (UC) were included in this prospective study. Blood and stool samples were collected before treatment (baseline). Serum hepcidin, hemoglobin levels, platelet counts, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), interleukin-6 (IL 6), ferritin, iron, soluble transferrin receptors, and fecal calprotectin were assessed. The same parameters were measured and compared with the baseline levels in the follow-up period, during maintenance therapy (average of 39 months after diagnosis). Results: Patients with CD (n=30) had higher serum hepcidin levels (expressed as a median and interquartile range) at diagnosis than subjects with UC (n=13). These levels significantly decreased during the follow-up (from 36.5 (11.5-79.6) ng/ml to 2.1 (0.9-6.7) ng/ml). Contrarily, no significant serum hepcidin level changes were observed in UC (from 5.4 (3.4-16.6) ng/ml to 4.8 (0.9-8.1) ng/ml). While in children with CD hepcidin level dynamics correlated with disease activity and inflammatory markers (ESR, CRP), an only correlation with serum iron levels was observed in patients with UC. Conclusion: Children with CD had higher serum hepcidin levels at diagnosis compared to subjects with UC. Decrease of serum hepcidin in the CD group during anti-inflammatory therapy has been observed, whereas low hepcidin levels in children with UC have remained unchanged. Acknowledgment: This study was supported by grant MH CZ–DRO (FNOl, 00098892).

Keywords: children, Crohn's disease, ulcerative colitis, anaemia, hepcidin

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Authors: Ziani Borhane Eddine Cherif, Hazzi Mohamed, Mouhouche Fazia

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The vegetal extracts of aromatic and medicinal plants start to have much of interest like potential sources of natural bioactive molecules. Many features are conferred by the nature of the chemical function of their major constituents (phenol, alcohol, aldehyde, cetone). This biopotential lets us to focalize on the study of three main biological activities, the antioxidant, antibiotic and insecticidal activities of six Algerian aromatic plants in the aim of making in evidence by the chromatographic analysis (CPG and CG/SM) the phytochemical compounds implicating in this effects. The contents of Oxygenated monoterpenes represented the most prominent group of constituents in the majority of plants. However, the α-Terpineol (28,3%), Carvacrol (47,3%), pulégone (39,5%), Chrysanthenone (27,4%), Thymol 23,9%, γ-Terpinene 23,9% and 2-Undecanone(94%) were the main components. The antioxyding activity of the Essential oils and no-volatils extracts was evaluated in vitro using four tests: inhibition of free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the 2,2-Azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) radical-scavenging activity (ABTS•+), the thiobarbituric acid reactive substances (TBARS) assays and the reducing power. The measures of the IC50 of these natural compounds revealed potent activity (between 254.64-462.76mg.l-1), almost similar to that of BHT, BHA, Tocopherol and Ascorbic acid (126,4-369,1 mg.l-1) and so far than the Trolox one (IC50= 2,82mg.l-1). Furthermore, three ethanol extracts were found to be remarkably effective toward DPPH and ABTS inhibition, compared to chemical antioxidant BHA and BHT (IC = 9.8±0.1 and 28±0.7 mg.l-1, respectively); for reducing power test it has also exhibited high activity. The study on the insecticidal activity effect by contact, inhalation, fecundity and fertility of Callosobruchus maculatus and Tribolium confusum showed a strong potential biocide reaching 95-100% mortality only after 24 hours. The antibiotic activity of our essential oils were evaluated by a qualitative study (aromatogramme) and quantitative (MIC, MBC and CML) on four bacteria (Gram+ and Gram-) and one strain of pathogenic yeast, the results of these tests showed very interesting action than that induced by the same reference antibiotics (Gentamycin, and Nystatin Ceftatidine) such that the inhibition diameters and MIC values for tested microorganisms were in the range of 23–58 mm and 0.015–0.25%(v/v) respectively.

Keywords: aromatic plants, essential oils, no-volatils extracts, bioactive molecules, antioxidant activity, insecticidal activity, antibiotic activity

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Authors: Rachmat Mauludin, Nurmazidah

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Background and purpose: Famotidine is an H2 receptor blocker. Absorption orally is rapid enough, but famotidine can be degraded by stomach acid causing dose reduction until 35.8% after 50 minutes. This drug also undergoes first-pass metabolism which reduced its bio availability only until 40-50%. To overcome these problems, Solid Lipid Nano particles (SLNs) as alternative delivery systems can be formulated. SLNs is a lipid-based drug delivery technology with 50-1000 nm particle size, where the drug incorporated into the bio compatible lipids and the lipid particles are stabilized using appropriate stabilizers. When the particle size is 200 nm or below, lipid containing famotidine can be absorbed through the lymphatic vessels to the subclavian vein, so first-pass metabolism can be avoided. Method: Famotidine SLNs with various compositions of stabilizer was prepared using a high-speed homogenization and sonication method. Then, the particle size distribution, zeta potential, entrapment efficiency, particle morphology and in vitro release profiles were evaluated. Optimization of sonication time also carried out. Result: Particle size of SLN by Particle Size Analyzer was in range 114.6 up to 455.267 nm. Ultrasonicated SLNs within 5 minutes generated smaller particle size than SLNs which was ultrasonicated for 10 and 15 minutes. Entrapment efficiency of SLNs were 74.17 up to 79.45%. Particle morphology of the SLNs was spherical and distributed individually. Release study of Famotidine revealed that in acid medium, 28.89 up to 80.55% of famotidine could be released after 2 hours. Nevertheless in basic medium, famotidine was released 40.5 up to 86.88% in the same period. Conclusion: The best formula was SLNs which stabilized by 4% Poloxamer 188 and 1 % Span 20, that had particle size 114.6 nm in diameter, 77.14% famotidine entrapped, and the particle morphology was spherical and distributed individually. SLNs with the best drug release profile was SLNs which stabilized by 4% Eudragit L 100-55 and 1% Tween 80 which had released 36.34 % in pH 1.2 solution, and 74.13% in pH 7.4 solution after 2 hours. The optimum sonication time was 5 minutes.

Keywords: famotodine, SLN, high speed homogenization, particle size, release study

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Authors: Ghibeche Abderrahmane

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To examine whether genes associated with cellular defense against oxidative stress are associated with insulin sensitivity, patients with type 2 diabetes (n=7) and age-matched (n=5) and young (n=9) control subjects underwent a euglycemic-hyperinsulinemic clamp for 120 min. Muscle samples were obtained before and after the clamp and analyzed for heat shock protein (HSP)72 and heme oxygenase (HO)-1 mRNA, intramuscular triglyceride content, and the maximal activities of β-hyroxyacyl-CoA dehydrogenase (β-HAD) and citrate synthase (CS). Basal expression of both HSP72 and HO-1 mRNA were lower (P < 0.05) by 33 and 55%, respectively, when comparing diabetic patients with age-matched and young control subjects, with no differences between the latter groups. Both basal HSP72 (r = 0.75, P < 0.001) and HO-1 (r = 0.50,P < 0.05) mRNA expression correlated with the glucose infusion rate during the clamp. Significant correlations were also observed between HSP72 mRNA and both β-HAD (r = 0.61, P < 0.01) and CS (r = 0.65, P < 0.01). HSP72 mRNA was induced (P < 0.05) by the clamp in all groups. Although HO-1 mRNA was unaffected by the clamp in both the young and age-matched control subjects, it was increased (P < 0.05) ∼70-fold in the diabetic patients after the clamp. These data demonstrate that genes involved in providing cellular protection against oxidative stress are defective in patients with type 2 diabetes and correlate with insulin-stimulated glucose disposal and markers of muscle oxidative capacity. The data provide new evidence that the pathogenesis of type 2 diabetes involves perturbations to the antioxidant defense mechanism within skeletal muscle.

Keywords: euglycemic-hyperinsulinemic, HSP72, mRNA, diabete

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Authors: Chiagoziem A. Otuechere, Ebenezer O. Farombi

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The protective effect of dichloromethane: methanol extract of Pterocarpus mildbraedii (PME), a widely consumed Nigerian leafy vegetable, on the toxicity of propanil was investigated in male rats. Animals were distributed into eight groups of five each. Group 1 served as control and received normal saline while rats in groups 2, 3, and 4 received 100 mg/kg, 200 mg/kg, and 400 mg/kg extract doses respectively. Group 5 rats were orally administered 200 mg/kg propanil while groups 6, 7, and 8 rats were given propanil plus extract. Oral administration of propanil elicited a 14.8%, 5%, 122%, and 78% increase in the activity of serum enzymes; alanine aminotransferase (AST), alanine aminotransferase(ALT), Alkaline phoshatase (ALP) and Gamma glutamyl transferase (ﻻGT). There were also increase in Lactate dehydrogenase (LDH) activity, direct bilirubin and lipid peroxidation levels. Furthermore, PME significantly attenuated the marked hepatic oxidative damage that accompanied propanil treatment. The extract significantly decreased LDH activity and bilirubin levels following propanil treatment. Furthermore, propanil-induced alterations in the activities of antioxidant enzymes: Superoxide dismutase (SOD), catalase (CAT) and glutathione s-transferase (GST) in these rats were modulated by the extract. The percentage DPPH Radical Scavenging Activity of the extract was determined as 55% and compared to those of Gallic acid (49%). Hepatic histology examination further confirmed the damage to the liver as it revealed severe periportal cellular infiltration of the hepatocytes. These biochemical and morphological alterations were attenuated in rats pre-treated with 100 mg/kg and 200 mg/kg doses of the extract. These results suggest that PME possesses protective effect against propanil-induced hepatotoxicity.

Keywords: antioxidant, hepatoprotection, Pterocarpus mildbraedii, propanil

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Authors: Seung Ju Yeon, Seul Ki Min, Jun Sang Park, Yeo Seon Kwon, Hoo Cheol Lee, Hyun Jung Shim, Il-Doo Kim, Ja Kyeong Lee, Hwa Sung Shin

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In brain injury, depleting oxidative stress is the most effective way to reduce the brain infarct size. C-phycocyanin (C-Pc) is a well-known antioxidant protein that has neuroprotective effects obtained from green microalgae. Astrocyte is glial cell that supports the nerve cell such as neuron, which account for a large portion of the brain. In brain injury, such as ischemia and reperfusion, astrocyte has an important rule that overcomes the oxidative stress and protect from brain reactive oxygen species (ROS) injury. However little is known about how C-Pc regulates the anti-oxidants effects of astrocyte. In this study, when the C-Pc was treated in oxidized astrocyte, we confirmed that inflammatory factors Interleukin-6 and Interleukin-3 were increased and antioxidants enzyme, Superoxide dismutase (SOD) and catalase was upregulated, and neurotrophic factors, brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) was alleviated. Also, it was confirmed to reduce infarct size of the brain in ischemia and reperfusion because C-Pc has anti-oxidant effects in middle cerebral artery occlusion (MCAO) animal model. These results show that C-Pc can help astrocytes lead neuroprotective activities in the oxidative stressed environment of the brain. In summary, the C-PC protects astrocytes from oxidative stress and has anti-oxidative, anti-inflammatory, neurotrophic effects under ischemic situations.

Keywords: c-phycocyanin, astrocyte, reactive oxygen species, ischemia and reperfusion, neuroprotective effect

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Authors: Sally A. El Awdan, Gehad A. Abdel Jaleel, Dalia O Saleh, Manal Badawi

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Background: Diabetes is a metabolic disease that affects a wide range of people worldwide and results in serious complications. Streptozotocin (STZ) causes selective cytotoxicity in the pancreatic β-cell, and it has been extensively used to induce diabetes mellitus in rats. The present study investigated the effects of diosmin and chrysin alone or in combination with each other on glucose level and on liver in STZ diabetic rats. Methods: In this study, rats were divided into six experimental groups (normal, untreated STZ-diabetic (60 mg/kg B.W., IP), treated STZ-diabetic with glycazide (10 mg/kg B.W, oral), treated STZ-diabetic with diosmin (100 mg/kg B. W., oral), treated STZ-diabetic with chrysin (80 mg/kg B.W., oral), treated STZ-diabetic with diosmin (50 mg/kg B.W, oral) + chrysin (40 mg/kg B.W., oral). After 2 weeks blood samples were withdrawn and glucose was measured. Animals were anaesthetized with an intraperitoneal injection of sodium pentobarbital (60 mg/kg), and sacrificed for dissecting liver. Results: Throughout the experimental period, all treatments significantly (P<0.05) lowered serum glucose, AST, ALT, triglyceride, cholesterol, IL-6, TNF-α and IL-1β. Moreover, the treated diabetic rats showed higher levels of reduced glutathione (P<0.05) in the liver compared to the diabetic control rats and inhibited diabetes-induced elevation in the levels of malondialdehyde in liver. The results of this study clearly demonstrated that diosmin and chrysin possess several treatment-oriented properties, including the control of hyperglycemia, antioxidant effects and anti-inflammatory effects. Conclusion: Considering these observations, it appears that diosmin and chrysin may be a useful supplement to delay the developmentof diabetes and its complications.

Keywords: diabetes, streptozocin, chrysin, rat, diosmin, cytokines

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Authors: A. Gramza-Michałowska, J. Skręty, M. Antczak, J. Kobus-Cisowska, D. Kmiecik, J. Korczak, Kulczyński Bartosz

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Background: Modern consumer highly appreciates the correlation between eating habits and health. Intensified research showed many proofs confirming that food besides its basic nutritional function, possess also significant prophylactic and therapeutic potential. Preventive potential of selected food is commonly used as improvement factor of patients life standard. World Health Organization indicates that diabetes (Diabetes mellitus) and obesity are two of the most common and dangerous diseases. Diet therapy is an element of diabetes education program and a part of healing process, allowing maintaining and remaining the optimal metabolic state of the system. It must be remembered that diabetes treatment should be individualized to each patient. One of highly recommended vegetable for diabetes is asparagus (Asparagus officinalis L.), low calories common plant, growing in European countries. Objective: To propose the technology of unsweetened muesli production with addition of new components, we investigated the effects of selected vegetable addition on antioxidative capacity and consumer’s acceptance of muesli as representative of breakfast product. Methods: Muesli was formulated from a composition of oat flakes, flaxseed, bran, carrots, broccoli and asparagus. Basic composition of muesli was evaluated as content of protein, lipids, fatty acid composition, ash, selected minerals and caloricity. Antioxidant capacity of muesli was evaluated with use radical scavenging methods (DPPH, ABTS), ORAC value and PCL - photochemiluminescence antiradical potential. Proposed muesli as new product was also characterized with sensory analysis, which included color, scent, taste, consistency and overall acceptance of a product. Results: Results showed that addition of freeze-dried asparagus into muesli allowed to lower the fat content and caloricity of a product according to the base product. No significant loss in antioxidant potential was evaluated, also the sensory value of a product was not negative. Conclusion: Designed muesli would be an answer for obese people looking for healthy snack during the daytime. Results showed that product with asparagus addition would be accepted by the consumers and because of its antidiabetic potential could be a n important factor in prevention of diabetes or obesity. Financial support by the UE Project no PO IG 01.01.00.00-061/09

Keywords: muesli, vegetables, asparagus, antioxidant potential, lipids

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Authors: Viktoriia Burkina, Vladimir Zlabek, Galia Zamaratskaia

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Cytochromes P450 (CYP) are important family of enzymes in Phase I metabolism. Environmental pollutants often act as inducers of the gene expression and activities CYP1A1 and CYP3A-like isoforms in fish. The activities are generally measured in the fish liver or gills, and less is known about tissue distribution of expression. In present study, the CYP1A1 and CYP3A-like activities were measured in rainbow trout liver, gill, intestine, heart, brain and gonads. The activities of CYP1A1 and CYP3A-like proteins were estimated as the rates of 7-ethoxyresorufin-O-deethylation (EROD) and benzyloxy-4-trifluoromethylcoumarin-O-debenzyloxylation (BFCOD), respectively. The CYP1A1 and CYP3A-like activities were detectable in all investigated fish tissues, with the highest activity in hepatic tissue followed by heart > brain > gill > intestine > gonads. To confirm the presence of CYP1A1 in different tissues, EROD activity was measured in presence of the selective inhibitors ellipticine (CYP1A1), ketoconazole (CYP3A), 8-methoxypsoralen (human CYP2A) and diallyl sulphide (CYP2E1). It was found that ellipticine, ketoconazole and 8-methoxypsoralen inhibited hepatic EROD activity by 88-98%. Ellipticine inhibited gill, intestine, and gonad EROD activity by 50%. In conclusion, EROD and BFCOD activities were detected in rainbow trout liver, gill, intestine, heart, brain and gonads. Further studies are needed to fully identify all CYP450 isoforms responsible for these activities. Acknowledgement: The study was financially supported by the Ministry of Education, Youth and Sports of the Czech Republic - projects „CENAKVA “(No. CZ.1.05/2.1.00/01.0024), “CENAKVA Center Development “(No. CZ.1.05/2.1.00/19.0380), “CENAKVA II “(No. LO1205 under the NPU I program), and "Development of USB - International mobility (No. CZ.02.2.69/0.0/0.0/16_027/0008364).

Keywords: BFCOD, EROD, fish, phase I metabolism, selective inhibitors

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Authors: Amelia R. Anshar, Dini Kurnia, Muh A. Bahar

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Nowadays, there are so many incidences had been reported in pet animals regarding drug overdose caused by incorrect doses of a non-steroidal anti-inflammatory drug (NSAID), for instance, meloxicam. As supporting treatment, the avocado is used in traditional medicine to treat or prevent some health cases. The study was aimed at providing the basis for the antioxidant activity of avocado puree in animal medicine. Experimental animals used in this study were 24 male rats that were randomly divided into 4 groups (n=6). Control Group I got 1 ml CMC 1% and control II got meloxicam 30 mg/kgBB and 1 ml CMC 1%. Treatment group I got meloxicam 30 mg/kgBB and avocado 5 g/kgBB/day and treatment II got meloxicam 30 mg/kgBB and avocado 10 g/kgBB/day. The study was conducted over 8 days, then the level of Blood Urea Nitrogen and creatinine of the white rats were examined in 1st day and 8th day. The results were analyzed by ANOVA Two Way With Replication, then followed by T-test (α = 0,05) if there were a difference. Comparison test among the four groups after treatment with avocado using Anova Two Way With Replication test showed that there were significant differences between the mean of the four groups either decreased levels of Blood Urea Nitrogen and creatinine with p < 0,05. Treatment group I and II received treatment showed remarkable (p < 0,05) decreases ini Blood Urea Nitrogen level by 27,17 mg/dl and 17,83 mg/dl respectively. There was also significant decrease in the values of creatinine in Treatment group I and treatment group II by 0,983 mg/dl and 0,713 mg/dl respectively. The conclusion of this study was that avocado decreases level of Blood Urea Nitrogen and creatinine in white rats which are exposed to toxic doses of meloxicam.

Keywords: avocado, blood urea nitrogen, creatinine, meloxicam

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Authors: Shrestha Sharma, Jasjeet K. Sahni, Javed Ali, Sanjula Baboota

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Introduction: Rutin is a naturally occurring strong antioxidant molecule belonging to bioflavonoid category. Due to its free radical scavenging properties, it has been found to be beneficial in the treatment of various diseases including inflammation, cancer, diabetes, allergy, cardiovascular disorders and various types of microbial infections. Despite its beneficial effects, it suffers from the problem of low aqueous solubility which is responsible for low oral bioavailability. The aim of our study was to optimize and characterize self-nanoemulsifying drug delivery system (SNEDDS) of rutin using Box-Behnken design (BBD) combined with a desirability function. Further various antioxidant, pharmacokinetic and pharmacodynamic studies were performed for the optimized rutin SNEDDS formulation. Methodologies: Selection of oil, surfactant and co-surfactant was done on the basis of solubility/miscibility studies. Sefsol+ Vitamin E, Solutol HS 15 and Transcutol P were selected as oil phase, surfactant and co-surfactant respectively. Optimization of SNEDDS formulations was done by a three-factor, three-level (33)BBD. The independent factors were Sefsol+ Vitamin E, Solutol HS15, and Transcutol P. The dependent variables were globule size, self emulsification time (SEF), % transmittance and cumulative percentage drug released. Various response surface graphs and contour plots were constructed to understand the effect of different factor, their levels and combinations on the responses. The optimized Rutin SNEDDS formulation was characterized for various parameters such as globule size, zeta potential, viscosity, refractive index , % Transmittance and in vitro drug release. Ex vivo permeation studies and pharmacokinetic studies were performed for optimized formulation. Antioxidant activity was determined by DPPH and reducing power assays. Anti-inflammatory activity was determined by using carrageenan induced rat paw oedema method. Permeation of rutin across small intestine was assessed using confocal laser scanning microscopy (CLSM). Major findings:The optimized SNEDDS formulation consisting of Sefsol+ Vitamin E - Solutol HS15 -Transcutol HP at proportions of 25:35:17.5 (w/w) was prepared and a comparison of the predicted values and experimental values were found to be in close agreement. The globule size and PDI of optimized SNEDDS formulation was found to be 16.08 ± 0.02 nm and 0.124±0.01 respectively. Significant (p˂0.05) increase in percentage drug release was achieved in the case of optimized SNEDDS formulation (98.8 %) as compared to rutin suspension. Furthermore, pharmacokinetic study showed a 2.3-fold increase in relative oral bioavailability compared with that of the suspension. Antioxidant assay results indicated better efficacy of the developed formulation than the pure drug and it was found to be comparable with ascorbic acid. The results of anti-inflammatory studies showed 72.93 % inhibition for the SNEDDS formulation which was significantly higher than the drug suspension 46.56%. The results of CLSM indicated that the absorption of SNEDDS formulation was considerably higher than that from rutin suspension. Conclusion: Rutin SNEDDS have been successfully prepared and they can serve as an effective tool in enhancing oral bioavailability and efficacy of Rutin.

Keywords: rutin, oral bioavilability, pharamacokinetics, pharmacodynamics

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Authors: Diego R. Boiarski, Camila M. Toigo, Thais M. Sobjak, Andrey F. P. Santos, Silvia Romao, Ana T. B. Guimaraes

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Anthropogenic activities promote changes in the community’s structures and decrease the species abundance of amphibians. Biological communities of fluvial systems are assemblies of organisms that have adapted to regional conditions, including the physical environment and food resources, and are further refined through interactions with other species. The aim of this study was to assess neurotoxic alterations and in the antioxidant system on tadpoles of Lithobates catesbeianus exposed to waters from Cascavel River, in the south of Brazil. A total of 420 L of water was collected from the Cascavel River, 140 L from each of the three different locations: Site 1 – headwater; Site 2 – stretch of the stream that runs through an urbanized area; Site 3 – a stretch from the rural area. Twelve tadpoles were acclimated in each aquarium (100 L of water) for seven days. The water from each aquarium was replaced with the ones sampled from the river, except the one from the control aquarium. After seven days, a portion of the liver was removed and conditioned for ChE, SOD, CAT and LPO analysis; other part of the tissue was conditioned for histological analysis. The statistical analysis performed was one-way ANOVA, followed by post-hoc Tukey-HSD test, and the multivariate principal components analysis. It was not observed any neurotoxic effect, but a slight increase in SOD activity and elevation of CAT activity in both urban and rural environment. A decrease in LPO reaction was detected, mainly among the tadpoles exposed to the waters from the rural area. The results of the present study demonstrate the alteration of the antioxidant system, as well as liver histopathologies in tadpoles exposed mainly to waters collected in urban and rural environments. These alterations may cause the reduction in the velocity of the metamorphosis process from the tadpoles. Further, were observed histological alterations, highlighting necrotic areas mainly among the animals exposed to urban waters. Those damages can lead to metabolic dysfunction, interfering with survival capacity, diminishing not only individual fitness but for the whole population. In the interpretation synthesis of all biomarkers, the cellular damage gradient is perceptible, characterized by the variables related to the antioxidant system, due to the flow direction of the stream. This result is indicative that along the course of the creek occurs dumping of organic material, which promoted an acute response upon tadpoles of L. catesbeianus. and it was also observed the difference in tissue damage between the experimental groups and the control group, the latter presenting histological alterations, but to a lesser degree than the animals exposed to the waters of the Cascavel river. These damages, caused by reactive oxygen species possibly resulting from the contamination by organic compounds, can lead the animals to a series of metabolic dysfunctions, interfering with its metamorphosis capacity. Interruption of metamorphosis may affect survival, which may impair its growth, development and reproduction, diminishing not only the fitness of each individual but in a long-term, to the entire population.

Keywords: American bullfrog, histopathology, oxidative stress, urban creeks pollution

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