Search results for: enzyme inhibition

Authors: Fengmei Li, Li Xu, Guoliang Xia

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Lysozyme is a catalytic enzyme that performs bacterial cell lysis by cleaving the β-1,4-glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine of peptidoglycan in cell walls. In the present study, an invertebrate-type (i-type) lysozyme gene was cloned from Chinese mitten crab Eriocheir sinensis (designated as EsLysozyme) based on PCR-based rapid amplification of cDNA ends (RACE) technology. The full-length cDNA of EsLysozyme was of 831 bp. SMART and SIGNALP 3.0 program analysis revealed that EsLysozyme contained a signal peptide and a destabilase domain. The five amino acid residues (Tyr63, Trp64, Tyr91, His110, Pro114) and the conserved motif GSLSCG(P/Y)FQI and CL(E/L/R/H)C(I/M)C in i-type lysozymes were also found in EsLysozyme. The high similarity of EsLysozyme with L. vannamei lysozymes and phylogenetic analysis suggested that EsLysozyme should be a new member of i-type lysozyme family.

Keywords: i-type lysozyme, Eriocheir sinensis, cloning, characterization

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Authors: Stephy Saavedra, Annsy C. Arredondo, Gisele Monteiro, Adalberto Pessoa Jr, Carol N. Flores-Fernandez, Amparo I. Zavaleta

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L-asparaginase from bacterial sources is used in leukemic treatment and food industry. This enzyme is classified based on its affinity towards L-asparagine and L-glutamine. Likewise, ansZ genes express L-asparaginase with higher affinity to L-asparagine. The aim of this work was to clone and express of ansZ gene from Bacillus sp. CH11 isolated from Chilca salterns in Peru. The gene encoding L-asparaginase was cloned into pET15b vector and transformed in Escherichia coli BL21 (DE3) pLysS. The expression was carried out in a batch culture using LB broth and 0.5 mM IPTG. The recombinant L-asparaginase showed a molecular weight of ~ 39 kDa by SDS PAGE and a specific activity of 3.19 IU/mg of protein. The cloning and expression of ansZ gene from this halotolerant Bacillus sp. CH11 allowed having a biological input to improve a future scaling-up.

Keywords: ansZ gene, Bacillus sp, Chilca salterns, recombinant L-asparaginase

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Authors: Yun-Hsuan Chang, Chih-Chun Huang, Ru-Band Lu

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Dopamine, metabolized to 3,4-dihydroxyphenylacetic acid (DOPAC) by aldehyde dehydrogenase 2 (ALDH2), ALDH2*1/*1, and ALDH2*1/*2+ALDH*2/*2 equally carried in Han Chinese. The relationship between dopamine metabolic enzyme and cognitive performance in bipolar II disorder comorbid with anxiety disorder (AD) remains unclear. This study proposed to explore the association between ALDH2 polymorphisms, anxiety comorbidity in bipolar II disorder. One hundred and ninety-seven BPII with or without AD comorbidity were recruited and compared with 130 Health controls (HC). A polymerase chain reaction and restriction fragment length polymorphism analysis was used to determine genotypes for ALDH2, and neuropsychological battery was performed. Two factor analyses with AD comorbidity and ALDH2 showed a significant main effect of ALDH2 on attention and marginally significant interaction between AD and ALDH2 memory performance. The ALDH2 polymorphisms may play a different role in the neuropsychological performance on varied neuropsychological performance in BPII comorbid with and without AD.

Keywords: anxiety disorder, bipolar II disorder, comorbidity, genetic

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Authors: Ivana Brzonova, Asina Fnu, Alena Kubatova, Evguenii Kozliak, Yun Ji

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Bio-decomposition of lignin by Basidiomycetes in the presence of dimethyl sulfoxide (DMSO) was investigated. The addition of 3-5 vol% DMSO to lignin aqueous media significantly increased the lignin solubility based on UV absorbance. After being dissolved in DMSO, the thermal evolution profile also changed significantly, yielding more high-MW organic carbon at the expense of recalcitrant elemental carbon. Medical fungi C. versicolor, G. lucidum and P. pulmonarius, were observed to grow on the lignin in media containing up to 15 vol. % DMSO. Further detailed product characterization by chromatographic methods corroborated these observations, as more low-MW phenolic products were observed with DMSO as a co-solvent. These results may be explained by the high solubility of lignin in DMSO; thus, the addition of DMSO to the medium increases the lignin availability for microorganisms. Some of these low-MW phenolic products host a big potential to be used in medicine. No significant inhibition of enzymatic activity (laccase, MnP, LiP) was observed by the addition of up to 3 vol% DMSO.

Keywords: basidiomycetes, bio-degradation, dimethyl sulfoxide, lignin

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Authors: Adferotimi Banso

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A laboratory investigation was conducted to determine the effect of Justica flava and Aframonium melegueta on the growth of Aspergillus niger, Rhizopus stolonifer and Fusarium species in sweet orange and mango juices. Aqueous extract (3%v/v) of Justica flava and Aframonium melegueta reduced the growth of the fungi, a combination of 2% (v/v) each of Justica flava and Aframonium melegueta extracts reduced the growth better. Partial purification of aqueous extracts of Justica flava and Aframonium melegueta showed that ethyl acetate fraction of the extracts exhibited the highest level of inhibition of growth of the test fungi compared with diethyl ether and n-hexane fractions. The results suggest that extracts of Justica flava and Aframonium melegueta may be important substitutes for conventional chemical preservatives in the processing of fruit juices.

Keywords: aqueous, fraction, mango, orange, purification, sweet

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Authors: M. A. Siti Faridah, K. Muhammad, H. M. Ghazali, Y. A. Yusof

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This study was conducted to investigate the effects of three variables namely types of cell wall degrading enzymes (Viscozyme L, Pectinex Ultra SP-L, Rapidase PAC, Rohament CL and Rohapect PTE) at varying concentrations (0.25-3% v/w) and times (30 min-24 h) on the zinc (Zn-) amaranth purees. Liquefaction treatment of the Zn-amaranth purees with Viscozyme (1% v/w at pH 5 and 45ºC for 3 h) was found to be the best procedure, which produced Zn-amaranth puree with low viscosity (8.60 mPas). Zn-amaranth purees were also found to have the highest metallo-chlorophyll derivative contents (0.16 mg/g), free radical 2, 2-diphenyl-1-picrylhydrazyl (DPPH) values (12.49 mM (TE)/g fresh weight) and ferric reducing antioxidant power (FRAP) values (4.57 mM (TE)/g fresh weight) within 3 h of liquefaction. Other physicochemical properties of the enzyme-liquefied Zn-amaranth purees indicated that lightness (L*) (12.54), greenness a*/b* (-0.30), reducing sugar (103.88 mg/mL) and soluble dietary fibre (5.94%) of the purees were higher compared to that of nonenzyme-liquefied amaranth purees.

Keywords: amaranth, antioxidant, chlorophyll derivative, enzymatic liquefaction

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Authors: Azna Zuberi

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Biofilm is a sessile bacterial accretion in which bacteria adapts different physiological and morphological behavior from planktonic form. It is the root cause of about 80% microbial infections in human. Among them, E. coli biofilms are most prevalent in medical devices associated nosocomial infections. The objective of this study was to inhibit biofilm formation by targeting LuxS gene, involved in quorum sensing using CRISPRi. luxS is a synthase, involved in the synthesis of Autoinducer-2(AI-2), which in turn guides the initial stage of biofilm formation. To implement CRISPRi system, we have synthesized complementary sgRNA to target gene sequence and co-expressed with dCas9. Suppression of luxS was confirmed through qRT-PCR. The effect of luxS gene on biofilm inhibition was studied through crystal violet assay, XTT reduction assay and scanning electron microscopy. We conclude that CRISPRi system could be a potential strategy to inhibit bacterial biofilm through mechanism base approach.

Keywords: biofilm, CRISPRi, luxS, microbial

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Authors: Asima Shah, F. A. Masoodi, Adil Gani, Bilal Ahmad Ashwar

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The present study was designed to evaluate the effect of γ-rays on the antioxidant and antiproliferative potential of β-glucan isolated from oats. The β-glucan was irradiated with 0, 2, 6, and 10 kGy by gamma ray. The samples were characterized by FT-IR, GPC, and quantitative estimation by Megazyme β-glucan assay kit. The average molecular weight of non-irradiated β-glucan was 199 kDa that decreased to 70 kDa at 10 kGy. Both FT-IR spectrum and chemical analysis revealed that the extracted β-glucan was pure having minor impurities. Antioxidant activity was evaluated by DPPH, lipid peroxidation, reducing power, metal chelating ability and oxidative DNA damage assays. Results revealed that the antioxidant activity of β-glucan increased with the increase in irradiation dose. Irradiated β-glucan also exhibited dose dependent cancer cell growth inhibition with irradiation doses. The study revealed that low molecular weight β-glucan with enhanced antioxidant and antiproliferative activities can be produced by a simple irradiation method.

Keywords: γ-irradiation, antioxidant activity, antiproliferative activity, β-glucan, oats

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Authors: Hamide Amani, Afshin FarahBakhsh, Iman Farahbakhsh

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In this paper, an amprometric biosensor based on surface modified polypyrrole (PPy) has been developed for the quantitative estimation of urea in aqueous solutions. The incorporation of urease (Urs) into a bipolymeric substrate consisting of PPy was performed by entrapment to the polymeric matrix, PPy acts as amperometric transducer in these biosensors. To increase the membrane conductivity, multi-walled carbon nanotubes (MWCNT) were added to the PPy solution. The entrapped MWCNT in PPy film and the bipolymer layers were prepared for construction of Pt/PPy/MWCNT/Urs. Two different configurations of working electrodes were evaluated to investigate the potential use of the modified membranes in biosensors. The evaluation of two different configurations of working electrodes suggested that the second configuration, which was composed of an electrode-mediator-(pyrrole and multi-walled carbon nanotube) structure and enzyme, is the best candidate for biosensor applications.

Keywords: urea biosensor, polypyrrole, multi-walled carbon nanotube, urease

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Authors: Margarita Ivanova, Julia Dao, Andrew Friedman, Neil Kasaci, Rekha Gopal, Ozlem Goker-Alpan

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In Fabry disease (FD), α-galactosidase A (α-Gal A) deficiency leads to the accumulation of globotriaosylceramide (Lyso-Gb3 and Gb3), triggering a pathologic cascade that causes the severity of organs damage. The heart is one of the several organs with high sensitivity to the α-Gal A deficiency. A subgroup of patients with significant residual of α-Gal A activity with primary cardiac involvement is occasionally referred to as “cardiac variant.” The cardiovascular complications are most frequently encountered, contributing substantially to morbidity, and are the leading cause of premature death in male and female patients with FD. The deposition of Lyso-Gb-3 and Gb-3 within the myocardium affects cardiac function with resultant progressive cardiovascular pathology. Gb-3 and Lyso-Gb-3 accumulation at the cellular level trigger a cascade of events leading to end-stage fibrosis. In the cardiac tissue, Lyso-Gb-3 deposition is associated with the increased release of inflammatory factors and transforming growth factors. Infiltration of lymphocytes and macrophages into endomyocardial tissue indicates that inflammation plays a significant role in cardiac damage. Moreover, accumulated data suggest that chronic inflammation leads to multisystemic FD pathology even under enzyme replacement therapy (ERT). NF-κB activation plays a subsequent role in the inflammatory response to cardiac dysfunction and advanced heart failure in the general population. TNFalpha/NF-κB signaling protects the myocardial evoking by ischemic preconditioning; however, this protective effect depends on the concentration of TNF-α. Thus, we hypothesize that TNF-α is a critical factor in determining the grade of cardio-pathology. Cardiac hypertrophy corresponds to the expansion of the coronary vasculature to maintain a sufficient supply of nutrients and oxygen. Coronary activation of angiogenesis and fibrosis plays a vital role in cardiac vascularization, hypertrophy, and tissue remodeling. We suggest that the interaction between the inflammatory pathways and cardiac vascularization is a bi-directional process controlled by secreted cytokines and growth factors. The co-coordination of these two processes has never been explored in FD. In a cohort of 40 patients with FD, biomarkers associated with inflammation and cardio hypertrophic remodeling were studied. FD patients were categorized into three groups based on LVmass/DSA, LVEF, and ECG abnormalities: FD with no cardio complication, FD with moderate cardio complication, and severe cardio complication. Serum levels of NF-kB, TNFalpha, Il-6, Il-2, MCP1, ING-gamma, VEGF, IGF-1, TGFβ, and FGF2 were quantified by enzyme-linked immunosorbent assays (ELISA). Among the biomarkers, MCP-1, INF-gamma, VEGF, TNF-alpha, and TGF-beta were elevated in FD patients. Some of these biomarkers also have the potential to correlate with cardio pathology in FD. Conclusion: The study provides information about the role of inflammatory pathways and biomarkers of cardio hypertrophic remodeling in FD patients. This study will also reveal the mechanisms that link intracellular accumulation of Lyso-GB-3 and Gb3 to the development of cardiomyopathy with myocardial thickening and resultant fibrosis.

Keywords: biomarkers, Fabry disease, inflammation, growth factors

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Authors: Manju Verma, Parag A. Deshpande

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Electronic structure calculations of hydrogen terminated metal-porphyrin graphene were carried out to explore the catalytic activity for CO2 hydration reaction. A ruthenium atom was substituted in place of carbon atom of graphene and ruthenium chelated carbon atoms were replaced by four nitrogen atoms in metal-porphyrin graphene system. Ruthenium atom created the active site for CO2 hydration reaction. Ruthenium-porphyrin graphene followed the mechanism of carbonic anhydrase enzyme for CO2 conversion to HCO3- ion. CO2 hydration reaction over ruthenium-porphyrin graphene proceeded via the elementary steps: OH- formation from H2O dissociation, CO2 bending in presence of nucleophilic attack of OH- ion, HCO3- ion formation from proton migration, HCO3- ion desorption by H2O addition. Proton transfer to yield HCO3- ion was observed as a rate limiting step from free energy landscape.

Keywords: ruthenium-porphyrin graphene, CO2 hydration, carbonic anhydrase, heterogeneous catalyst, density functional theory

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Authors: Ali Fadhel Ahmed, Tuqa Abdulkareem Hameed

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Moringa oleifera (leaves and seeds) ethanolic and aqueous extracts were tested for antibacterial activity. The effect of plant extracts on three types of bacterial species: Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae, was investigated. Using the agar well diffusion method, ethanolic extracts of Moringa oleifera demonstrated a significant antibacterial effect on the forty tested bacterial strains. Seed-induced inhibition zones (ethanolic extracts)were ranged from16 to 24 mm in diameter against S. aureus, respectively, whileE. coli and K. pneumonia had no effect. Gram-positive and Gram-negative bacteria were not affected by alcoholic and aqueous plant leaf extracts. The purpose of this present study was to look at the cytotoxic effects of M.Oleifera plant (alcoholic extracts).

Keywords: moringa oleifera, escherichia coli, klebsiella pneumoniae, staphylococcus aureus

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Authors: Gülşah Kanbur, Veysel Ayhan

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Feed oil samples which are used as mixed feed raw material were taken from six different feed factories in March, May and July. All factories make production in Konya, Turkey and all of the samples were which taken are crude soybean oil. Some physical and chemical analysis, free radical scavenger effect and total phenol content were determined on these oil samples. Moisture content was found between 0.10-22.23 %, saponification number was determined 143.13 to 167.93 KOH/kg, free fatty acidity was varied 0.73 to 35.00 % , peroxide value was found between 1.53 and 28.43 meq/kg , unsaponifiable matter was determined from 0.40 to 17.10 % , viscosity was found between 34.30 and 625.67 mPas, sediment amount was determined between 0.60-18.16 % , free radical scavenger effect was varied 20.7 to 43.04 % inhibition of the extract and total phenol content was found between 1.20 and 2.69 mg/L extract. Different results were found between months and factories.

Keywords: crude soybean oil, feed oils, mixed feed, Konya

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Authors: Ben Amar Cheba, Taha Ibrahim Zaghloul, Mohamad Hisham El-Massry, Ahmad Rafik El-Mahdy

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Seventy two bacterial strains with ability to degrade chitin were isolated during a screening program. One of the most potent isolates (strain R2) was identified as Bacillus sp. using conventional methods as well as 16S rRNA technique and submitted in the Gen Bank sequence database as Bacillus sp. R2 with a given accession number DQ 923161. This strain was able to produce high levels of extracellular chitinase. The chitinase of Bacillus sp. R2 hydrolyzed several chitinous substrates preferentially and showed a maximum activity toward the β chitin such as Calmar pen and squid bone chitins with the folds 1.47 and 1.23 respectively. The enzyme also exhibited a substrate binding capacity of more than 70% for squid chitin, shrimp shell colloidal chitin, chitosan and prawn shell chitin. The chitinase showed a moderate antifungal activity against many phytopathogenic fungi such as Aspergillus niger, A. flavus, Penicillium degitatum and Fusarium calmorum.This strain could be a suitable candidate for chitinase production on an industrial scale for using as promising antifungal biopestecide.

Keywords: antifungal activity, Bacillus sp. R2, chitinase, substrate specificity

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Authors: Boudjelal Amel

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Arisarum vulgare, a member of the Araceae family, is an herbaceous perennial widely distributed in the Mediterranean region. A. vulgare is recognized for its medicinal properties and holds significant traditional importance in Algeria for the treatment of various human ailments, including pain, infections, inflammation, digestive disorders, skin problems, eczema, cancer, wounds, burns and gynecological diseases. Despite its extensive traditional use, scientific exploration of A. vulgare remains limited. The study aims to investigate for the first time the therapeutic potential of A. vulgare ethanolic extract obtained by ultrasound-assisted extraction. The chemical composition of the extract was determined by LC-MS/MS analysis. For in vitro phytopharmacological evaluation, several assays, including DPPH, ABTS, FRAP and reducing power, were employed to evaluate the antioxidant activity. The antibacterial activity was assessed againt Escherichia coli, Salmonella typhimurium, Staphylococus aureus, Enterococcus feacium by disk diffusion and microdilution methods. The possible inhibitory activity of ethanolic extract was analyzed against the cholinesterases enzymes (AChE and BChE). The DNA protection activity of A. vulgare ethanolic extract was estimated using the agarose gel electrophoresis method. The capacities of the extract to protect plasmid DNA (pBR322) from the oxidizing effects of H2O2 and UV treatment were evaluated by their DNA-breaking forms. The in vivo wound healing potential of a traditional ointment containing 5% of A. vulgare ethanolic extract was also investigated. The LC-MS/MS profiling of the extract revealed the presence of various bioactive compounds, including naringenin, chlorogenic, vanillic, cafeic, coumaric acids, trans-cinnamic and trans ferrulic acids. The plant extract presented considerable antioxidant potential, being the most active for Reducing power (0,07326±0.001 mg/ml) and DPPH (0.14±0.004 mg/ml). The extract showed the highest inhibition zone diameter against Enterococcus feacium (36±0.1 mm). The ethanolic extract of A. vulgare suppressed the growth of Staphylococus aureus, Escherichia coli and Salmonella typhimurium according to the MIC values. The extract of the plant significantly inhibited both AChE and BChE enzymes. DNA protection activity of the A. vulgare extract was determined as 90.41% for form I and 51.92% for form II. The in vivo experiments showed that 5% ethanolic extract ointment accelerated the wound healing process. The topical application of the traditional formulation enhanced wound closure (95,36±0,6 %) and improved histological parameters in the treated group compared to the control groups. The promising biological properties of Arisarum vulgare revealed that the plant could be appraised as a potential origin of bioactive molecules having multifunctional medicinal uses.

Keywords: arisarum vulgare, LC-MS/MS, antioxidant activity, antimicrobial activity, cholinesterases enzymes inhibition, dna-damage activity, in vivo wound healing

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Authors: Kyungbae Pi, Kibeom Lee, Yongil Kim, Eun-Jung Lee

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Abnormal adipocyte growth, in terms of increased cell numbers and increased cell differentiation, is considered to be a major pathological feature of obesity. Thus, the inhibition of preadipocyte mitogenesis and differentiation could help prevent and suppress obesity. The aim of this study was to assess whether extracts from Weissella koreensis 521 cells isolated from kimchi could exert anti-adipogenic effects in 3T3-L1 cells (fat cells). Differentiating 3T3-L1 cells were treated with W. koreensis 521 cell extracts (W. koreensis 521_CE), and cell viability was assessed by MTT assays. At concentrations below 0.2 mg/ml, W. koreensis 521_CE did not exert any cytotoxic effect in 3T3-L1 cells. However, treatment with W. koreensis 521_CE significantly inhibited adipocyte differentiation, as assessed by morphological analysis and Oil Red O staining of fat. W. koreensis 521_CE treatment (0.2 mg/ml) also reduced lipid accumulation by 24% in fully differentiated 3T3-L1 adipocytes. These findings collectively indicate that Weissella koreensis 521 may help prevent obesity.

Keywords: Weissella koreensis 521, 3T3-L1 cells, adipocyte differentiation, obesity

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Authors: M. Rajasekar, K. Suresh

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Diosmin is a flavonoid, most abundantly found in many citrus fruits. As a flavonoid, it possesses a multitude of biological activities including anti-hyperglycemic, anti-lipid peroxidative, anti-inflammatory, antioxidant, and anti-mutagenic properties. At this point, we established the anti-proliferative and apoptosis-inducing activities of diosmin in Hep-2 and KB cells. Diosmin has cytotoxic effects through inhibiting cellular proliferation of Hep-2 and KB cells, which leads to the induction of apoptosis, as apparent by an increase in the fraction of cells in the sub-G1phase of the cell cycle. Results exposed that inhibition of cell proliferation is associated with regulation of the Interleukins/STAT pathway. In addition, Diosmin treatment with Hep-2 and KB cells actively stimulated reactive oxygen species (ROS) and mitochondrial membrane depolarization. And also an imbalance in the Bax/Bcl-2 ratio triggered the caspase cascade and shifting the balance in favor of apoptosis. These observations conclude that Diosmin induce apoptosis via Interleukins /STAT-mediated pathway.

Keywords: diosmin, apoptosis, antioxidant, STAT pathway

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Authors: Rezvan Najafi, Korosh Heydari, Massoud Saidijam

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5-FU is a chemotherapeutic agent that has been used in colorectal cancer (CRC) treatment. However, it is usually associated with the acquired resistance, which decreases the therapeutic effects of 5-FU. miR-200c is involved in chemotherapeutic drug resistance, but its mechanism is not fully understood. In this study, the effect of inhibition of miR-200c in sensitivity of HCT-116 CRC cells to 5-FU was evaluated. HCT-116 cells were transfected with LNA-anti- miR-200c for 48 h. mRNA expression of miR-200c was evaluated using quantitative real- time PCR. The protein expression of phosphatase and tensin homolog (PTEN) and E-cadherin were analyzed by western blotting. Annexin V and propidium iodide staining assay were applied for apoptosis detection. The caspase-3 activation was evaluated by an enzymatic assay. The results showed LNA-anti-miR-200c inhibited the expression of PTEN and E-cadherin protein, apoptosis and activation of caspase 3 compared with control cells. In conclusion, these results suggest that miR-200c as a prognostic marker can overcome to 5-FU chemoresistance in CRC.

Keywords: colorectal cancer, miR-200c, 5-FU resistance, E-cadherin, PTEN

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Authors: Yazun Jarrar

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Smoking has effect on platelet aggregation and the activity of anti-platelet drugs. The chemical 20-hydroxyeicosatetraenoic acid (20-HETE) is a cardiotoxic arachidonic acid metabolite which increases platelet aggregation. In this study, we investigated the influence of cigarette smoking on 20-HETE levels and protein expression of 20-HETE producing enzyme CYP4A11 in isolated platelets from smoker and non-smoker volunteers. The protein expression and 20-HETE levels were analyzed using immunoblot and High-Performance Liquid Chromatography with Mass Spectrometry (HPL-MS) assays. The results showed that 20-HETE level was higher significantly among smokers than non-smokers (t-test, p-value<0.05). The protein expression of CYP4A11 was significantly higher (t-test, p-value<0.05) among the platelets of smokers. We concluded that cigarette smoking increased the level of platelet activator 20-HETE through increasing the protein expression of CYP4A11. These findings may increase the understanding of smoking-drug interaction during antiplatelets therapy.

Keywords: smoking, 20-HETE, CYP4A11, platelet

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Authors: Rahma Tibigui, Ikram Hadj Said, Rachid Belkada, Dalila Hammoutene

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The vulnerability of industrial facilities is highly concerned with multiple risks from corrosion. The commonly adopted solution is based on the use of organic inhibitors, which are gradually being replaced by environmentally friendly organic inhibitors. In our work, we carried out a quantum chemical study based on the Density Functional Theory (DFT) method at the B3LYP/6-311G (d,p) level of theory. The inhibitory performance of a derivative of the tetrazole molecule has been investigated and reported as a carbon steel-friendly corrosion inhibitor in hydrochloric acid (HCl) medium. The relationship is likely to exist between the molecular structure of this compound as well as its various global reactivity descriptors, and its corrosion inhibition efficiency, which was examined and then discussed. The results show low values of ΔE, which represent strong adsorption of the inhibitor on the steel surface. Moreover, the flat adsorption orientation confirmed the great ability to donate (accept) electrons to (from) steel, fabricating an anchored barrier to prevent steel from corrosion.

Keywords: eco-friendly, corrosion inhibitors, tetrazole, DFT

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Authors: Tesfaye Alamirew, Delele Worku, Solomon W. Fanta, Nigus Gabbiye

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Recently graphen based nanocomposites are become an emerging research areas for fabrication of enzymatic biosensors due to their property of large surface area, conductivity and biocompatibility. This review summarizes recent research reports of graphen based nanocomposites for the fabrication of glucose and ethanol enzymatic biosensors. The newly fabricated enzyme free microwave treated nitrogen doped graphen (MN-d-GR) had provided highest sensitivity towards glucose and GCE/rGO/AuNPs/ADH composite had provided far highest sensitivity towards ethanol compared to other reported graphen based nanocomposites. The MWCNT/GO/GOx and GCE/ErGO/PTH/ADH nanocomposites had also enhanced wide linear range for glucose and ethanol detection respectively. Generally, graphen based nanocomposite enzymatic biosensors had fast direct electron transfer rate, highest sensitivity and wide linear detection ranges during glucose and ethanol sensing.

Keywords: glucose, ethanol, enzymatic biosensor, graphen, nanocomposite

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Authors: A. Abdenbi, B. Dennai, B. Touati, M. Bouaaza, A. Saad

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The essential oils of many plants have become popular in recent years and their bioactive principles have recently won several industry sectors, however their use as antibacterial and anti fungal agents has been reported. This study focuses on the physico chemical and phyto chemical with a study of the antimicrobial activity of essential oils of aromatic and medicinal plant of southwest Algeria, this essential oil was obtained by hydro-distillation of aerial parts of Cotula cinerea, belonging to the Asteraceae family, it is very extensive in the spring season in a region called Kenadza road, located 12km from Bechar. Variable anti fungal activity of the essential oil of Cotula cinerea (yield 2%) were revealed about four fungal strains, the minimum inhibitory concentrations of essential oils were determined by the method of dilution in agar. Significant fungal sensitivity of Penicillium sp with an inhibition of 32.3 mm area.

Keywords: Cotula cinerea, essential oil, physico- chemical analysis and phyto- chemical, anti fungal power

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Authors: Mohamad Shurair, Fares Almomani, Simon Judd, Rahul Bhosale, Anand Kumar, Ujjal Gosh

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This study evaluated the use of mixed indigenous microalgae (MIMA) as a treatment process for wastewaters and CO2 capturing technology at different temperatures. The study follows the growth rate of MIMA, removals of organic matter, removal of nutrients from synthetic wastewater and its effectiveness as CO2 capturing technology from flue gas. A noticeable difference between the growth patterns of MIMA was observed at different CO2 and different operational temperatures. MIMA showed the highest growth grate when injected with CO2 dosage of 10% and limited growth was observed for the systems injected with 5% and 15 % of CO2 at 30 ◦C. Ammonia and phosphorus removals for Spirulina were 69%, 75%, and 83%, and 20%, 45%, and 75% for the media injected with 0, 5 and 10% CO2. The results of this study show that simple and cost-effective microalgae-based wastewater treatment systems can be successfully employed at different temperatures as a successful CO2 capturing technology even with the small probability of inhibition at high temperatures

Keywords: greenhouse, climate change, CO2 capturing, green algae

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Authors: Kimberley Phoena Fan, Yu Zhang

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ECOSURF™ EH-3 Surfactant (EH3) is a nonionic surfactant and has superior wetting and excellent oil removal properties. It is biodegradable with low toxicity and meets or exceeds US EPA Design for the Environment Criteria, and is widely used as a home cleaner, commercial and industrial degreaser. We have recently found that EH3 also possesses a special function which is characterized as an enhancer to Taq DNA polymerase and ameliorator to reduce the effects of PCR inhibitors, i.e., blood, urea, Guanidinium thiocyanate, Humic acids, polyphenol, and Polysaccharides. This is a new kind of PCR enhancer that does not work on relieving secondary structures of GC-rich templates. We have compared EH3’s effects on Taq DNA Polymerase along with other well-known enhancers, such as DMSO, betaine, and BSA, using GC rich or deficient template and found that, unlike DMSO and Betaine, the EH3 boosting effect on PCR reaction is not through reducing Tm. The results show the same increase of PCR products regardless of the GC contents or secondary structures. The mechanism of EH3 enhancing PCR is through its direct interaction with or stimulation of the DNA polymerase and making the enzymes more resistant to inhibitors in the presence of EH3. This phenomenon has first been observed for EH3, a new type of PCR enzyme enhancer. Subsequent research also shows that a series of similar surfactants boost Taq DNA polymerase as well.

Keywords: EH3, DNA, polymerase, enhancer, raw biological samples

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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: Ayodeji O. Falade, Leonard V. Mabinya, Uchechukwu U. Nwodo, Anthony I. Okoh

Abstract:

Given the high utility of peroxidase in several industrial processes, the search for novel microorganisms with enhanced peroxidase production capacity is of keen interest. This study investigated the process conditions for optimum peroxidase production by Ensifer sp, new ligninolytic proteobacteria with peroxidase production potential. Also, some agricultural residues were valorized for peroxidase production under solid state fermentation. Peroxidase production was optimum at an initial medium pH 7, incubation temperature of 30 °C and agitation speed of 100 rpm using alkali lignin fermentation medium supplemented with guaiacol as the most effective inducer and ammonium sulphate as the best inorganic nitrogen. Optimum peroxidase production by Ensifer sp. was attained at 48 h with specific productivity of 12.76 ± 1.09 U mg⁻¹. Interestingly, probable laccase production was observed with optimum specific productivity of 12.76 ± 0.45 U mg⁻¹ at 72 h. The highest peroxidase yield was observed with sawdust as solid substrate under solid state fermentation. In conclusion, Ensifer sp. possesses the capacity for enhanced peroxidase production that can be exploited for various biotechnological applications.

Keywords: catalase-peroxidase, enzyme production, peroxidase, polymerase chain reaction, proteobacteria

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Authors: Hye-Hyun Lee, Hyo-Kon Chun, Kyung-Hee Kim Kim, Mi-Hee Kim, Saet-Byul Chu, Sang-Jong Lee, Seung-Hyeop Lee, Seung-Won Yi

Abstract:

Some chemicals such as malachite green, methylene blue, and copper sulfate had been used frequently as disinfectants controlling fungal infection in aquaculture. However, their carcinogenicity, mutagenicity and teratogenicity were reported in mammals. After their accumulation in food fish and its consumers was confirmed, concerns about public health has resulted in enhanced monitoring and increased demand for eco-friendly treatments. Therefore, this study aimed to evaluate safety to fish and efficacy of sulfur solution processed by effective microorganisms (EM-PSS) against Saprolegnia parasitica, for use of a potential aquatic fungicidal disinfectant. The natural sulfur purchased from Kawah Ijen volcano, East Java, Indonesia was processed by the liquid mixture consisting of following twelve effective microorganisms (Rapha-el®; Lbiotech, Jeonnam, Korea), Lactobacillus parafarraginis, L. paracasei, L. harbinensis, L. buchneri, L. perolens, L. rhamnosus, L. vaccinostercus, Acetobacter lovaniensis, A. peroxydans, Pichia fermentans, Candida ethanolica, Saccharomycopsis schoenii isolated from fermentation process of oriental medicinal herbs including green tea, privet, and puer tea. The material was applied to in vitro antifungal activity test for Saprolegnia parasitica using agar dilution method. In addition, an acute toxicity test was performed on carp (Cyprinus carpio), eel (Anguilla japonica), and mud fish (Misgurnus mizolepis) for 96 hours. After three species of fish (n=15) were accustomed to experimental water environment for three days, the EM-PSS was added to each tank as final concentrations to be 0 to 500 ppm. The fish were taken into necropsy, and the histological sections of the gill, liver, and spleen were counter-stained with hematoxylin and eosin (H-E). And hence, no observed effect concentration (NOEC) of the solution was used for taking a medicinal bath for mudfish infected by Saprolegnia parasitica in practice. The result of in vitro antifungal activity test showed the growth inhibition of the fungus at 100 ppm, which and the lower concentrations occurred no fatal case in any fish species tested until the end of the examination. The 125 ppm of the solution, however, resulted in 13.3 %, 13.3 %, and 6.3 % of mortality in carp, eel, and mudfish, respectively. But both 250 and 500 ppm of the solution leaded lethality to all population of each fish species within 24 hours. Besides, H-E staining also showed no specific evidence for toxicity in fish at lesser than 100 ppm of EM-PSS. On the other hand, as a result of field application of the solution, no growth of fungal mycelium was found in fish bodies from gross observation 5 days post treatment. In conclusion, 100ppm of EM-PSS resulted in inhibition and treatment of Saprolegnia parasitica infection. In addition, the use of EM-PSS lower than 100 ppm is safe for fish. Therefore, EM-PSS could be used as aquatic fungicide, and also may be possible to be a potential eco-friendly disinfectant in aquaculture.

Keywords: antifungal activity, effective microorganism, toxicity, saprolegnia, processed sulfur solution

Procedia PDF Downloads 249

Authors: Debora B. Moretti, Wiolene M. Nordi, Thaline Maira P. Cruz, José Eurico P. Cyrino, Raul Machado-Neto

Abstract:

Enzyme activity was evaluated in the intestine of juvenile dourado (Salminus brasiliensis) fed with diets containing 0, 10 or 20% of lyophilized bovine colostrum (LBC) inclusion for either 30 or 60 days. The intestinal enzymes acid and alkaline phosphatase (ACP and ALP, respectively), non-specific esterase (NSE), lipase (LIP), dipeptidyl aminopeptidase IV (DAP IV) and leucine aminopeptidase (LAP) were studied using histochemistry in four intestinal segments (S1, S2, S3 and posterior intestine). Weak proteolitic activity was observed in all intestinal segments for DAP IV and LAP. The activity of NSE and LIP was also weak in all intestines, except for the moderate activity of NSE in the S2 of 20% LBC group after 30 days and in the S1 of 0% LBC group after 60 days. The ACP was detected only in the S2 and S3 of the 10% LBC group after 30 days. Moderate and strong staining was observed in the first three intestinal segments for ALP and weak activity in the posterior intestine. The activity of DAP IV, LAP and ALP were also present in the cytoplasm of the enterocytes. In the present results, bovine colostrum feeding did not cause alterations in activity of intestinal enzymes.

Keywords: carnivorous fish, enterocyte, intestinal epithelium, teleost

Procedia PDF Downloads 320

Authors: Rashad Al-Hindi

Abstract:

The study aimed at investigating and screening of some contaminants in some honey-based products. Sixty-nine 'honey paste' samples marketed in Jeddah, Saudi Arabia, were subjected to physicochemical studies and screening of aflatoxins and pesticide residues. The physicochemical parameters studied were mainly: moisture content, total sugars, total ash, total nitrogen, fibres, total acidity as citric acid and pH. These parameters were investigated using standard methods of analysis. Mycotoxins (aflatoxins) and pesticide residues were by an enzyme-linked immunosorbent assay (ELISA) according to official methods. Results revealed that mean values of the examined criteria were: 15.44±0.36%; 74±4.30%; 0.40±0.062%; 0.22±0.05%; 6.93±1.30%; 2.53±0.161 mmol/kg; 4.10±0.158, respectively. Overall results proved that all tested honey pastes samples were free from mycotoxins (aflatoxins) and pesticide residues. Therefore, we conclude that 'honey pastes' marketed in Jeddah city, Saudi Arabia were safe for human consumption.

Keywords: aflatoxins, honey mixtures, pesticide residues, physicochemical

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Authors: Biswabandhu Bankura, Srikanta Guria, Madhusudan Das

Abstract:

Purpose: Thyroid peroxidase (TPO) is the key enzyme in the biosynthesis of thyroid hormones. We aimed to identify the spectrum of mutations in the TPO gene leading to hypothyroidism in the population of West Bengal to establish the genetic etiology of the disease. Methods: 200 hypothyroid patients (case) and their corresponding sex and age matched 200 normal individuals (control) were screened depending on their clinical manifestations. Genomic DNA was isolated from peripheral blood samples and TPO gene (Exon 7 to Exon 14) was amplified by PCR. The PCR products were subjected to sequencing to identify mutations. Results: Single nucleotide changes such as Glu 641 Lys, Asp 668 Asn, Thr 725 Pro, Asp 620 Asn, Ser 398 Thr, and Ala 373 Ser were found. Changes in the TPO were assayed in vitro to compare mutant and wild-type activities. Five mutants were enzymatically inactive in the guaiacol and iodide assays. This is a strong indication that the mutations are present at crucial positions of the TPO gene, resulting in inactivated TPO. Key Findings: The results of this study may help to develop a genetic screening protocol for goiter and hypothyroidism in the population of West Bengal.

Keywords: thyroid peroxidase, hypothyroidism, mutation, in vitro assay, transfection

Procedia PDF Downloads 334