Search results for: alternative oxidase

Authors: Khaled S. Al Salhen

Abstract:

Aldehyde oxidase is molybdo-flavoenzyme involved in the oxidation of hundreds of endogenous and exogenous and N-heterocyclic compounds and environmental pollutants. Uncharged N-heterocyclic aromatic compounds such phenanthridine are commonly distributed pollutants in soil, air, sediments, surface water and groundwater, and in animal and plant tissues. Phenanthridine as uncharged N-heterocyclic aromatic compound was incubated with partially purified aldehyde oxidase from rainbow trout fish liver. Reversed-phase HLPC method was used to separate the oxidation products from phenanthridine and the metabolite was identified. The 6(5H)-phenanthridinone was identified the major metabolite by partially purified aldehyde oxidase from fish liver. Kinetic constant for the oxidation reactions were determined spectrophotometrically and showed that this substrate has a good affinity (Km = 78 ± 7.6 µM) for hepatic aldehyde oxidase, coupled with a relatively high oxidation rate (0.77± 0.03 nmol/min/mg protein). In addition, the kinetic parameters of hepatic fish aldehyde oxidase towards the phenanthridine substrate indicate that in vitro biotransformation by hepatic fish aldehyde oxidase will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase.

Keywords: aldehyde oxidase, fish, phenanthridine, specificity

Procedia PDF Downloads 334

Authors: Kyung-Soon Choi, Ji-Young Hwang, Young-Hee Pyo

Abstract:

Vinegars have been used as an alternative remedy for treating gout, but the scientific basis remains to be elucidated. In this study, acetic acid fermentation was applied for the first time to Monascus-fermented soybeans to examine its effect on the bioactive components together with the xanthine oxidase inhibitory (XOI) activity of the soy vinegar. The content of total phenols (0.47~0.97 mg gallic acid equivalents/mL) and flavonoids (0.18~0.39 mg quercetin equivallents/mL) were spectrophotometrically determined, and the content of organic acid (10.22~59.76 mg/mL) and isoflavones (6.79~7.46 mg/mL) were determined using HPLC-UV. The analytical method for ubiquinones (0.079~0.276 μg/mL) employed saponification before solvent extraction and quantification using LC-MS. Soy vinegar also showed significant XOI (95.3%) after 20 days of acetic acid fermentation at 30 °C. The results suggest that soy vinegar has potential as a novel medicinal food.

Keywords: acetic acid fermentation, bioactive component, soy vinegar, xanthine oxidase inhibitory activity

Procedia PDF Downloads 351

Authors: Rahile Ozturk, Esra Maltas

Abstract:

This study aimed to determine the effect of vinclozolin on some biochemical characteristics of Galleria mellonella (Lepidoptera: Pyralidae) which is an economically harmful species damaging the honeycomb in beekeeping. For experimental groups, the eggs obtained from stock were dropped into the mixed feed of vinclozolin at different doses (20, 40 and 60 ppm) and had the larvae fed with this feed. As result of the addition of vinclozolin at concentrations of 20, 40 and 60 ppm, glycogen contents of G. mellonella were determined and a significant reduction in the amount of glycogen was observed with increasing concentration of vinclozolin. In this study, activity of catalase enzyme, particularly effective in defense mechanism, activity of xanthine oxidase involved in nucleotide metabolism and activity of glucose oxidase in the metabolism of carbohydrates were measured. When compared with the results from control groups, the enzyme activities of the larvaes fed with the feed including 20, 40 and 60 ppm of vinclozolin were observed to vary or remain constant. Accordingly, glucose oxidase and catalase activities increased with the increase in amount of vinclozolin in the feed and the activity of xanthine oxidase remained stable.

Keywords: Catalase, Galleria mellonella, glucose oxidase, vinclozolin, xanthine oxidase.

Procedia PDF Downloads 267

Authors: Farideh K. Khosroushahi, Serkan Erdal, Mucip Geni̇şel

Abstract:

Soil salinity is one of the major abiotic stress factors that restricts arable land and reduces crop productivity worldwide. High salt concentration adversely affects plant growth and development inducing water deficit, ionic toxicity, nutrient imbalance, and lead to oxidative stress. Although the stimulating role of mammalian sex hormones on various biological and biochemical processes under normal and stress condition have been proven, there is no study regarding with these hormone's effect on modulation of the alternative respiration pathway and AOX gene expression. In this study, changes in alternative respiration pathway in leaves of maize seedlings under salinity and the possible modulating effect of estrogen on these changes were investigated. Maize seedlings were grown in a hydroponic media for 11 days and then were exposed to salt stress for 3 days after being sprayed estrogen. The data obtained from oxygen consumption revealed that salt stress elevated cellular respiration value in the leaves. In addition, a marked increase was observed at alternative respiration level in salt-stressed seedlings. Compared to salt application alone, supplementation with estrogen resulted in a significant rise in alternative oxidase (AOX) activities. Similarly, while salt stress caused to rise in expressions of AOX gene compared to control seedlings, estrogen application resulted in further activation of these genes’ expression compared to stressed-seedlings alone. These data revealed that mitigating role of estrogen against the detrimental effects of salt stress is linked to modulation of alternative respiration pathway.

Keywords: alternative oxidase, estrogen, Ssalt stress, AOX, maize

Procedia PDF Downloads 179

Authors: M. S. Kilic, S. Korkut, B. Hazer

Abstract:

Newly synthesized Polypropylene-g-Polyethylene glycol polymer was first time used for a compartment-less enzymatic fuel cell. Working electrodes based on Polypropylene-g-Polyethylene glycol were operated as unmediated and mediated system (with ferrocene and gold/cobalt oxide nanoparticles). Glucose oxidase and bilirubin oxidase was selected as anodic and cathodic enzyme, respectively. Glucose was used as fuel in a single-compartment and membrane-less cell. Maximum power density was obtained as 0.65 nW cm-2, 65 nW cm-2, and 23500 nW cm-2 from the unmediated, ferrocene and gold/cobalt oxide modified polymeric film, respectively. Power density was calculated to be ~16000 nW cm-2 for undiluted wastewater sample with gold/cobalt oxide nanoparticles including system.

Keywords: bilirubin oxidase, enzymatic fuel cell, glucose oxidase, nanoparticles

Procedia PDF Downloads 228

Authors: Seyhan Sahan-Firat, Meryem Temiz-Resitoglu, Demet Sinem Guden, Sefika Pinar Kucukkavruk, Bahar Tunctan, Ayse Nihal Sari, Zumrut Kocak

Abstract:

We hypothesized that mTOR inhibition may prevent the multiple organ failures following severe multiple tissue injury associated with increased NADPH oxidase system activity occur in zymosan-induced systemic inflammation. Therefore, we investigated the role of mTOR in oxidative/nitrosative stress associated with increase in NADPH oxidase activity in zymosan-induced systemic inflammation model in rats. Male Wistar rats received saline (4 ml/kg, i.p.) and zymosan (500 mg/kg, i.p.) at time 0. Saline, or zymosan-treated rats were given rapamycin (1 mg/kg, i.p.) 1 h after saline or zymosan injections. Rats were sacrified 4 h after zymosan challenge and kidney, heart, thoracic aorta, and superior mesenteric artery were collected. NADPH oxidase activity, p22phox, gp91phox, and p47phox protein expression and nitrotyrosine levels were measured in tissue samples. Zymosan administration caused an increase in NADPH oxidase activity, p22phox, gp91phox, and p47phox protein expression and nitrotyrosine levels in kidney, heart, thoracic aorta, and superior mesenteric artery. These changes caused by zymosan reversed by rapamycin, a selective mTOR inhibitor. Rapamycin alone had no effect on the parameters measured. Our results demonstrated that zymosan-induced oxidative/nitrosative stress presumably due to enhanced activity of NADPH oxidase, expression of p22phox, gp91phox, and p47phox and production of peroxynitrite were mediated by mTOR. [This work was financially supported by Research Foundation of Mersin University (2016-2-AP3-1900)].

Keywords: oxidative stress, mTOR, nitrosative stress, zymosan

Procedia PDF Downloads 285

Authors: Jung-Feng Hsieh, Chu Ze Chen

Abstract:

Curcumin is the main active compound of turmeric that can inhibit the activities of α-glucosidase and xanthine oxidase (XO). α-Glucosidase and XO inhibitors are widely used to treat patients with diabetes mellitus and gout, respectively; therefore, the objective of this research was to evaluate the inhibitory activities of curcumin and its analogs against α-glucosidase and XO. Our results demonstrated that CM-F had the strongest antioxidant activity with a half-maximal effective concentration (EC50) of 9.39 ± 0.16 μM, which was superior to vitamin E (EC50=17.03 ± 0.09 μM). CM-F also exhibited potent inhibitory activity against XO with an IC50 value of 6.14 ± 0.38 μM and enzyme kinetic results revealed competitive inhibition of XO. We also found that CM-1 and CM-2 inhibited α-glucosidase with IC50 values of 21.06 ± 0.92 μM and 5.95 ± 0.09 μM, respectively, and kinetic studies indicated that both CM-1 and CM-2 are mixed competitive inhibitors of α-glucosidase. Furthermore, docking simulation identified five hydrogen bonds between XO and CM-F; however, only one and two hydrogen bonds are involved in CM-1 and CM-2 binding to α-glucosidase, respectively. Accordingly, curcumin and its analogs have the potential to be used in the treatment of patients with diabetes mellitus and gout.

Keywords: curcumin, α-glucosidase, inhibitor, xanthine oxidase

Procedia PDF Downloads 171

Authors: Maria Ostroukhova, Zhanneta Zalutskaya, Elena Ermilova

Abstract:

The alternative oxidase (AOX) mediates cyanide-resistant respiration, which bypasses proton-pumping complexes III and IV of the cytochrome pathway to directly transfer electrons from reduced ubiquinone to molecular oxygen. In Chlamydomonas reinhardtii, AOX is a monomeric protein that is encoded by two genes of discrete subfamilies, AOX1 and AOX2. Although AOX has been proposed to play essential roles in stress tolerance of organisms, the role of subfamily AOX2 is largely unknown. In C. reinhardtii, AOX2 was initially identified as one of constitutively low expressed genes. Like other photosynthetic organisms C. reinhardtii cells frequently experience periods of hypoxia. To examine AOX2 transcriptional regulation and role of AOX2 in hypoxia adaptation, real-time PCR analysis and artificial microRNA method were employed. Two experimental approaches have been used to induce the anoxic conditions: dark-anaerobic and light-anaerobic conditions. C. reinhardtii cells exposed to the oxygen deprivation have shown increased AOX2 mRNA levels. By contrast, AOX1 was not an anoxia-responsive gene. In C. reinhardtii, a subset of genes is regulated by transcription factor CRR1 in anaerobic conditions. Notable, the AOX2 promoter region contains the potential motif for CRR1 binding. Therefore, the role of CRR1 in the control of AOX2 transcription was tested. The CRR1-underexpressing strains, that were generated and characterized in this work, exhibited low levels of AOX2 transcripts under anoxic conditions. However, the transformants still slightly induced AOX2 gene expression in the darkness. These confirmed our suggestions that darkness is a regulatory stimulus for AOX genes in C. reinhardtii. Thus, other factors must contribute to AOX2 promoter activity under dark-anoxic conditions. Moreover, knock-down of CRR1 caused a complete reduction of AOX2 expression under light-anoxic conditions. These results indicate that (1) CRR1 is required for AOX2 expression during hypoxia, and (2) AOX2 gene is regulated by CRR1 together with yet-unknown regulatory factor(s). In addition, the AOX2-underexpressing strains were generated. The analysis of amiRNA-AOX2 strains suggested a role of this alternative oxidase in hypoxia adaptation of the alga. In conclusion, the results reported here show that C. reinhardtii AOX2 gene is stress inducible. CRR1 transcriptional factor is involved in the regulation of the AOX2 gene expression in the absence of oxygen. Moreover, AOX2 but not AOX1 functions under oxygen deprivation. This work was supported by Russian Science Foundation (research grant № 16-14-10004).

Keywords: alternative oxidase 2, artificial microRNA approach, chlamydomonas reinhardtii, hypoxia

Procedia PDF Downloads 210

Authors: R. D. Esposito, V. M. Barberá, P. García Morales, P. Dorado Rodríguez, J. Sanz, M. Fuentes, D. Planes Meseguer, M. Saceda, L. Fernández Fornos, M. P. Ventero

Abstract:

Results obtained by our group on glioblastoma multiforme (GBM) primary cultures , show a dramatic potentiation of radiation effects when 2 units/ml of D-amino acid oxidase (DAO) enzyme are added, free or immobilized in magnetic nanoparticles, to irradiated samples just after the irradiation. Cell cultures were exposed to radiation doses of 7Gy and 15Gy of 6 MV photons from a clinical linear accelerator. At both doses, we observed a clear enhancing effect of radiation-induced damages due to the addition of DAO.

Keywords: D-amino Acid Oxidase (DAO) enzyme, magnetic particles, nanotechnology, radiation therapy enhancement

Procedia PDF Downloads 488

Authors: Uc-Cayetano E. G., Ake-Uh O. E., Villanueva-Mena I. E., Ordonez L. C.

Abstract:

Carbon nanotubes (CNTs) and other graphitic nanostructures are materials with extraordinary physical, physicochemical and electrochemical properties which are being aggressively investigated for a variety of sensing applications. Thus, sensing of biological molecules such as proteins, DNA, glucose and other enzymes using either single wall or multiwall carbon nanotubes (MWCNTs) has been widely reported. Despite the current progress in this area, the electrochemical response of CNTs used in a variety of sensing arrangements still needs to be improved. An alternative towards the enhancement of this CNTs' electrochemical response is to chemically (or physically) modify its surface. The influence of the decoration with iron oxide nanoparticles in different types of MWCNTs on the amperometric sensing of glucose, urea, and cholesterol in solution is investigated. Commercial MWCNTs were oxidized in acid media and subsequently decorated with iron oxide nanoparticles; finally, the enzymes glucose oxidase, urease, and cholesterol oxidase are chemically immobilized to oxidized and decorated MWCNTs for glucose, urease, and cholesterol electrochemical sensing. The results of the electrochemical characterizations consistently show that the presence of iron oxide nanoparticles decorating the surface of MWCNTs enhance the amperometric response and the sensitivity to increments in glucose, urease, and cholesterol concentration when compared to non-decorated MWCNTs.

Keywords: WCNTs, enzymes, oxidation, decoration

Procedia PDF Downloads 93

Authors: Pawasuth Saengdee, Chamras Promptmas, Ting Zeng, Silke Leimkühler, Ulla Wollenberger

Abstract:

Sulfite has been used as a versatile preservative to limit the microbial growth and to control the taste in some food and beverage. However, it has been reported to cause a wide spectrum of severe adverse reactions. Therefore, it is important to determine the amount of sulfite in food and beverage to ensure consumer safety. An efficient electrocatalytic biosensor for sulfite detection was developed by immobilizing of human sulfite oxidase (hSO) on 3-aminoproplytriethoxysilane (APTES) modified indium tin oxide (ITO) electrode. Cyclic voltammetry was employed to investigate the electrochemical characteristics of the hSO modified ITO electrode for various pretreatment and binding conditions. Amperometry was also utilized to demonstrate the current responses of the sulfite sensor toward sodium sulfite in an aqueous solution at a potential of 0 V (vs. Ag/AgCl 1 M KCl). The proposed sulfite sensor has a linear range between 0.5 to 2 mM with a correlation coefficient 0.972. Then, the additional polymer layer of PVA was introduced to extend the linear range of sulfite sensor and protect the enzyme. The linear range of sulfite sensor with 5% coverage increases from 2.8 to 20 mM at a correlation coefficient of 0.983. In addition, the stability of sulfite sensor with 5% PVA coverage increases until 14 days when kept in 0.5 mM Tris-buffer, pH 7.0 at 4 8C. Therefore, this sensor could be applied for the detection of sulfite in the real sample, especially in food and beverage.

Keywords: sulfite oxidase, bioelectrocatalytsis, indium tin oxide, direct electrochemistry, sulfite sensor

Procedia PDF Downloads 191

Authors: Bhavini Patel, Aslihan Ugun-Klusek, Ellen Billet

Abstract:

The two main contributing factors to familial and idiopathic form of Parkinson’s disease (PD) are oxidative stress and altered proteolysis. Monoamine oxidase-A (MAO-A) plays a significant role in redox homeostasis by producing reactive oxygen species (ROS) via deamination of for example, dopamine. The ROS generated induces chemical modification of proteins resulting in altered biological function. The ubiquitin-proteasome system, which consists of three different types or proteolytic activity, namely “chymotrypsin-like” activity (CLA), “trypsin-like” activity (TLA) and “post acidic-like” activity (PLA), is responsible for the degradation of ubiquitinated proteins. Defects in UPS are known to be strongly correlated to PD. Herein, the effect of ROS generated by MAO-A on proteasome activity and the effects of proteasome inhibition on MAO-A protein levels in WT, mock and MAO-A overexpressed (MAO-A+) SHSY5Y neuroblastoma cell lines were investigated. The data in this study report increased proteolytic activity when MAO-A protein levels are significantly increased, in particular CLA and PLA. Additionally, 20S proteasome inhibition induced a decrease in MAO-A levels in WT and mock cells in comparison to MAO-A+ cells in which 20S proteasome inhibition induced increased MAO-A levels to be further increased at 48 hours of inhibition. This study supports the fact that MAO-A could be a potential pharmaceutical target for neuronal protection as data suggests that endogenous MAO-A levels may be essential for modulating cell death and survival.

Keywords: monoamine oxidase, neurodegeneration, Parkinson's disease, proteasome

Procedia PDF Downloads 96

Authors: Naouel Boussoualim, Hayat Trabsa, Imane Krache, Seddik Khennouf, Noureddine Charef, Lekhmici Arrar, Abderrahmane Baghiani

Abstract:

Purpose: To evaluate the in-vitro inhibition of xanthine oxidase (purified from bovine milk) by extracts of Lycium arabicum, as well as it is in vivo hypouricemic and renal protective effects. Methods: Four extracts of Lycium arabicum, methanol (CrE), chloroform (ChE), ethyl acetate (EaE) and aqueous (AqE) extracts, were screened for their total phenolics and potential inhibitory effects on purified bovine milk xanthine oxidase (XO) activity by measuring the formation of uric acid or superoxide radical. The mode of inhibition was investigated and compared with the standard drugs, allopurinol, quercitin, and catechin. To evaluate their hypouricemic effect, the extracts were administered to potassium oxonate-induced hyperuricemic mice at a dose of 50 mg/kg body weight. Results: The results showed that EaE had the highest content of phenolic compounds and was the most potent inhibitor of uric acid formation (IC50 = 0.017 ± 0.001 mg/mL) and formation of superoxide (IC50 = 0.035 ± 0.001 mg/ml). Lineweaver-Burk analysis showed that CrE and EaE inhibited XO competitively, whereas the inhibitory activities exerted by ChE and AqE were of a mixed type. Intraperetoneal injection of L. arabicum extracts (50 mg/kg) elicited hypouricemic actions in hyperuricemic mice. Hyperuricemic mice presented a serum uric acid concentration of 4.71 ± 0.29 mg/L but this was reduced to 1.78 ± 0.11 mg/L by EaE, which was the most potent hyporuricemic extract. Conclusion: L. arabicum fractions have a strong inhibitory effect on xanthine oxidase and and also have a significantly lowering effect on serum and liver creatinine and urea levels in hyperuricemic mice.

Keywords: lycium arabicum, uric acid, creatinine, superoxide, phenolic compounds, flavonoids, hyperuricemia

Procedia PDF Downloads 359

Authors: Arleta Rifati Nixha, Mustafa Arslan, Adem Ergün, Nahit Gencer

Abstract:

Polyphenol oxidase (PPO), sometimes referred to as phenol oxidase, catecholase, phenolase, catechol oxidase, or even tyrosinase, is considered to be an o-dipenol. PPO (EC 1.14.18.1), a multifunctional copper containing enzyme, is widely distributed in nature. It catalyzes two distinct reactions of melanin synthesis: a hydroxylation of monophenols to o-diphenols (monophenolase activity) and an oxidation of o-diphenols to o-quinones (diphenolase activity), both using molecular oxygen. Additionaly, investigation demonstrated that various dermatological disorders, such as age spots and freckle, were caused by the accumulation of an excessive level of epidermal pigmentation. Tyrosinase has also been linked to Parkinson’s and other neurodegenerative diseases. Nitrogen heterocycles have received a great deal of attention in the literature because of biological properties. Especially, among these heterocyclic systems, pyridine containing compounds have been the subject of expanding research efforts in heteroaromatic and biological chemistry. The pyrido [2,3-d] pyrimidine heterocycles, which are those annelated to a pyrimidine ring, are important because of their wide range of biological and pharmaceutical applications (i.e., bronchodilators, vasodilators) and their anti-allergic, cardiotonic, antihypertensive, and hepatoprotective activities. In this study series of 12 new carbazole substituted pyridopyrimidine urea(thiourea) derivatives were synthesized and evaluated effect on PPO. Additionally, we presented structure-activity relationship analyses and theoretical calculations of the compounds.

Keywords: carbazole, pyridopyrimidine, urea, thiourea, tyrosinase inhibitors

Procedia PDF Downloads 398

Authors: Gulnur Arabaci

Abstract:

Heavy metals are one of the major groups of contaminants in the environment and many of them are toxic even at very low concentration in plants and animals. However, some metals play important roles in the biological function of many enzymes in living organisms. Metals such as zinc, iron, and cooper are important for survival and activity of enzymes in plants, however heavy metals can inhibit enzyme which is responsible for defense system of plants. Polyphenol oxidase (PPO) is a copper-containing metalloenzyme which is responsible for enzymatic browning reaction of plants. Enzymatic browning is a major problem for the handling of vegetables and fruits in food industry. It can be increased and effected with many different futures such as metals in the nature and ground. In the present work, PPO was isolated and characterized from green leaves of red poppy plant (Papaver rhoeas). Then, the effect of some known antibrowning agents which can form complexes with metals and metals were investigated on the red poppy PPO activity. The results showed that glutathione was the most potent inhibitory effect on PPO activity. Cu(II) and Fe(II) metals increased the enzyme activities however, Sn(II) had the maximum inhibitory effect and Zn(II) and Pb(II) had no significant effect on the enzyme activity. In order to reduce the effect of heavy metals, the effects of metal-antibrowning agent complexes on the PPO activity were determined. EDTA and metal complexes had no significant effect on the enzyme. L-ascorbic acid and metal complexes decreased but L-ascorbic acid-Cu(II)-complex had no effect. Glutathione–metal complexes had the best inhibitory effect on Red poppy leaf PPO activity.

Keywords: inhibition, metal, red poppy, poly phenol oxidase (PPO)

Procedia PDF Downloads 287

Authors: Antonella Cartoni, Mattea Carmen Castrovilli

Abstract:

A biosensor for lactate detection has been developed using an environmentally friendly approach. The biosensor is based on lactate oxidase (LOX) and has remarkable capabilities for reuse and storage at room temperature. The manufacturing technique employed is ambient electrospray deposition (ESD), which enables efficient and sustainable immobilization of the LOX enzyme on a cost-effective com-mercial screen-printed Prussian blue/carbon electrode (PB/C-SPE). The study demonstrates that the ESD technology allows the biosensor to be stored at ambient pressure and temperature for extended periods without affecting the enzymatic activity. The biosensor can be stored for up to 90 days without requiring specific storage conditions, and it can be reused for up to 24 measurements on both freshly prepared electrodes and electrodes that are three months old. The LOX-based biosensor exhibits a lin-ear range of lactate detection between 0.1 and 1 mM, with a limit of detection of 0.07±0.02 mM. Ad-ditionally, it does not exhibit any memory effects. The immobilization process does not involve the use of entrapment matrices or hazardous chemicals, making it environmentally sustainable and non-toxic compared to current methods. Furthermore, the application of a electrospray deposition cycle on previously used biosensors rejuvenates their performance, making them comparable to freshly made biosensors. This highlights the excellent recycling potential of the technique, eliminating the waste as-sociated with disposable devices.

Keywords: green friendly, reuse, storage performance, immobilization, matrix-free, electrospray deposition, biosensor, lactate oxidase, enzyme

Procedia PDF Downloads 27

Authors: Yasser M. Saad, Heba El-Sebaie Abd El-Sadek

Abstract:

Some Metapenaeus monoceros cox1 gene fragments were isolated, purified, sequenced, and comparatively analyzed with some other Crustacean Cox1 gene sequences (obtained from National Center for Biotechnology Information). This work was designed for testing the efficiency of this system in reconstruction of phylogenetic relations among some Crustacean species belonging to four genera (Metapenaeus, Artemia, Daphnia and Calanus). The single nucleotide polymorphism and haplotype diversity were calculated for all estimated mt-DNA fragments. The genetic distance values were 0.292, 0.015, 0.151, and 0.09 within Metapenaeus species, Calanus species, Artemia species, and Daphnia species, respectively. The reconstructed phylogenetic tree is clustered into some unique clades. Cytochrome oxidase subunit 1 gene (cox1) was a powerful system in reconstruction of phylogenetic relations among evaluated crustacean species.

Keywords: crustaceans, genetics, Cox1, phylogeny

Procedia PDF Downloads 324

Authors: Navodit Goel, Prabir K. Paul

Abstract:

Tomato (Solanum lycopersicum) is attacked by Pseudomonas syringae pv. tomato causing speck lesions on the leaves leading to severe economic casualty. In the present study, aqueous fruit extracts of Azadirachta indica (neem) were sprayed on a single node of tomato plants grown under controlled contamination-free conditions. The treatment of plants was performed with neem fruit extract either alone or along with the pathogen. The parameters of observation were activities of polyphenol oxidase (PPO) and lysozyme, and isoform analysis of PPO; both at the treated leaves as well as untreated leaves away from the site of extract application. Polyphenol oxidase initiates phenylpropanoid pathway resulting in the synthesis of quinines from cytoplasmic phenols and production of reactive oxygen species toxic to broad spectrum microbes. Lysozyme is responsible for the breakdown of bacterial cell wall. The results indicate the upregulation of PPO and lysozyme activities in both the treated and untreated leaves along with de novo expression of newer PPO isoenzymes (which were absent in control samples). The appearance of additional PPO isoenzymes in bioelicitor-treated plants indicates that either the isoenzymes were expressed after bioelicitor application or the already expressed but inactive isoenzymes were activated by it. Lysozyme activity was significantly increased in the plants when treated with the bioelicitor or the pathogen alone. However, no new isoenzymes of lysozyme were expressed upon application of the extract. Induction of resistance by neem fruit extract could be a potent weapon in eco-friendly plant protection strategies.

Keywords: Azadirachta indica, lysozyme, polyphenol oxidase, Solanum lycopersicum

Procedia PDF Downloads 250

Authors: Anahita Izadyar

Abstract:

Using a recombinant enzyme derived from corn and a simple modification, we are fabricating a facile, fast, and cost-beneficial novel biosensor to measure glucose. We are applying Plant Produced Mn Peroxidase (PPMP), glucose oxidase (GOx), polyaniline (PANI) as conductive polymer and gold nanoparticles (AuNPs) on Au electrode using electrochemical response to detect glucose. We applied the entrapment method of enzyme composition, which is generally used to immobilize conductive polymer and facilitate electron transfer from the enzyme oxidation-reduction center to the sample solution. In this work, the oxidation of glucose on the modified gold electrode was quantified with Linear Sweep Voltammetry(LSV). We expect that the modified biosensor has the potential for monitoring various biofluids.

Keywords: plant-produced manganese peroxidase, enzyme-based biosensors, glucose, modified gold nanoparticles electrode, polyaniline

Procedia PDF Downloads 160

Authors: Ayesha Imtiaz, Darlina Md. Naim

Abstract:

Family Nemipteridae is among the most abundantly distributed family in Malaysian fish markets due to its high contribution to landing sites of Malaysia. Using an advanced molecular approach that used two mitochondrial (Cytochrome oxidase c I and Cytochrome oxidase b) and one nuclear gene (Recombination activating gene, RAGI) to expose cryptic diversity and phylogenetic relationships among commercially important species of family Nemipteridae. Our research covered all genera (including 31 species out total 45 species) of family Nemipteridae, distributed in Malaysia. We also found certain type of geographical barriers in the South China sea that reduces dispersal and stops a few species to intermix. Northside of the South China Sea (near Vietnam) does not allow genetic diversity to mix with the Southern side of the South China sea (Sarawak) and reduces dispersal. Straits of Malacca reduce the intermixing genetic diversity of South China Sea and the Indian Ocean.

Keywords: Nemipteridae, RAG I, south east Asia, Malaysia

Procedia PDF Downloads 109

Authors: M. Omer Faruk, A. M. A. M. Zonaed Siddiki, M. Fazal Karim, Md. Masuduzzaman, S. Chowdhury, Md. Shafiqul Islam, M. Alamgir Hossain

Abstract:

The dog tapeworms Echinococcus granulosus develop hydatid cysts in various organs in human and domestic animals worldwide including Bangladesh. The aim of this study was to identify and characterize the genotype of E. granulosus isolated from cattle using 12S rRNA and Cytochrome oxidase 1 (COX 1) genes. A total of 43 hydatid cyst samples were collected from 390 examined cattle samples derived from slaughterhouses. Among them, three cysts were fertile. Genomic DNA was extracted from germinal membrane and/or protoscoleces followed by PCR amplification of mitochondrial 12S rRNA and Cytochrome oxidase 1 gene fragments. The sequence data revealed existence of G1 (64.28%) and possible G3 (21.43%) genotypes for the first time in Bangladesh. The study indicates that common sheep strain G1 is the dominant subtype of E. granulosus in Chittagong region of Bangladesh. This will increase our understanding of the epidemiology of hydatidosis in the southern part of the country and will be useful to plan suitable control measures in the long run.

Keywords: Echinococcus granulosus, Cox1, 12S rRNA, molecular characterization, Bangladesh

Procedia PDF Downloads 303

Authors: Michal Kielbik, Izabela Szulc-Kielbik, Anna Brzostek, Jaroslaw Dziadek, Magdalena Klink

Abstract:

The goal of many research groups in the world is to find new components that are important for survival of mycobacteria in the host cells. Mycobacterium tuberculosis (Mtb) possesses a number of enzymes degrading cholesterol that are considered to be an important factor for its survival and persistence in host macrophages. One of them - cholesterol oxidase (ChoD), although not being essential for cholesterol degradation, is discussed as a virulence compound, however its involvement in macrophages’ response to Mtb is still not sufficiently determined. The recognition of tubercle bacilli antigens by pathogen recognition receptors is crucial for the initiation of the host innate immune response. An important receptor that has been implicated in the recognition and/or uptake of Mtb is Toll-like receptor type 2 (TLR2). Engagement of TLR2 results in the activation and phosphorylation of intracellular signaling proteins including IRAK-1 and -4, TRAF-6, which in turn leads to the activation of target kinases and transcription factors responsible for bactericidal and pro-inflammatory response of macrophages. The aim of these studies was a detailed clarification of the role of Mtb cholesterol oxidase as a virulence factor affecting the TLR2 signaling pathway in human macrophages. As human macrophages the THP-1 differentiated cells were applied. The virulent wild-type Mtb strain (H37Rv), its mutant lacking a functional copy of gene encoding cholesterol oxidase (∆choD), as well as complimented strain (∆choD–choD) were used. We tested the impact of Mtb strains on the expression of TLR2-depended signaling proteins (mRNA level, cytosolic level and phosphorylation status). The cytokine and bactericidal response of THP-1 derived macrophages infected with Mtb strains in relation to TLR2 signaling pathway dependence was also determined. We found that during the 24-hours of infection process the wild-type and complemented Mtb significantly reduced the cytosolic level and phosphorylation status of IRAK-4 and TRAF-6 proteins in macrophages, that was not observed in the case of ΔchoD mutant. Decreasement of TLR2-dependent signaling proteins, induced by wild-type Mtb, was not dependent on the activity of proteasome. Blocking of TLR2 expression, before infection, effectively prevented the induced by wild-type strain reduction of cytosolic level and phosphorylation of IRAK-4. None of the strains affected the surface expression of TLR2. The mRNA level of IRAK-4 and TRAF-6 genes were significantly increased in macrophages 24 hours post-infection with either of tested strains. However, the impact of wild-type Mtb strain on both examined genes was significantly stronger than its ΔchoD mutant. We also found that wild-type strain stimulated macrophages to release high amount of immunosuppressive IL-10, accompanied by low amount of pro-inflammatory IL-8 and bactericidal nitric oxide in comparison to mutant lacking cholesterol oxidase. The influence of wild-type Mtb on this type of macrophages' response strongly dependent on fully active IRAK-1 and IRAK-4 signaling proteins. In conclusion, Mtb using cholesterol oxidase causes the over-activation of TLR2 signaling proteins leading to the reduction of their cytosolic level and activity resulting in the modulation of macrophages response to allow its intracellular survival. Supported by grant: 2014/15/B/NZ6/01565, National Science Center, Poland

Keywords: Mycobacterium tuberculosis, cholesterol oxidase, macrophages, TLR2-dependent signaling pathway

Procedia PDF Downloads 389

Authors: Jyoti Kumari, Pavuluri Srinivasa Rao

Abstract:

Jackfruit (ArtocarpusheterophyllusL.) is an underutilized yet highly nutritious fruit with unique flavour, known for its therapeutic and culinary properties. Fresh jackfruit bulb has a very short shelf life due to high moisture and sugar content leading to microbial and enzymatic browning, hindering its consumer acceptability and marketability. An attempt has been made for the preservation of the ripe jackfruit bulbs, by the application of high pressure (HP) over a range of 200-500 MPa at ambient temperature for dwell times ranging from 5 to 20 min. The physicochemical properties of jackfruit bulbs such as the pH, TSS, and titrable acidity were not affected by the pressurization process. The ripening index of the fruit bulb also decreased following HP treatment. While the ascorbic acid and antioxidant activity of jackfruit bulb were well retained by high pressure processing (HPP), the total phenols and carotenoids showed a slight increase. The HPP significantly affected the colour and textural properties of jackfruit bulb. High pressure processing was highly effective in reducing the browning index of jackfruit bulbs in comparison to untreated bulbs. The firmness of the bulbs improved upon the pressure treatment with longer dwelling time. The polyphenol oxidase has been identified as the most prominent oxidative enzyme in the jackfruit bulb. The enzymatic activity of polyphenol oxidase and peroxidase were significantly reduced by up to 40% following treatment at 400 MPa/15 min. HPP of jackfruit bulbs at ambient temperatures is shown to be highly beneficial in improving the shelf stability, retaining its nutrient profile, color, and appearance while ensuring the maximum inactivation of the spoilage enzymes.

Keywords: antioxidant capacity, ascorbic acid, carotenoids, color, HPP-high pressure processing, jackfruit bulbs, polyphenol oxidase, peroxidase, total phenolic content

Procedia PDF Downloads 136

Authors: Sangeeta Palekar, Neeraj Rangwani, Akash Poddar, Jayu Kalambe

Abstract:

The bio-medical analysis is an indispensable procedure for identifying health-related diseases like diabetes. Monitoring the glucose level in our body regularly helps us identify hyperglycemia and hypoglycemia, which can cause severe medical problems like nerve damage or kidney diseases. This paper presents a method for predicting the glucose concentration in blood samples using image processing and machine learning algorithms. The glucose solution is prepared by the glucose oxidase (GOD) and peroxidase (POD) method. An experimental database is generated based on the colorimetric technique. The image of the glucose solution is captured by the raspberry pi camera and analyzed using image processing by extracting the RGB, HSV, LUX color space values. Regression algorithms like multiple linear regression, decision tree, RandomForest, and XGBoost were used to predict the unknown glucose concentration. The multiple linear regression algorithm predicts the results with 97% accuracy. The image processing and machine learning-based approach reduce the hardware complexities of existing platforms.

Keywords: artificial intelligence glucose detection, glucose oxidase, peroxidase, image processing, machine learning

Procedia PDF Downloads 163

Authors: Khaled S. Al Salhen

Abstract:

Aldehyde oxidase (AO) and xanthine oxidoreductase (XOR) catalyze the oxidation of many different N-heterocyclic compounds as well as aliphatic and aromatic aldehydes to their corresponding lactam and carboxylic acids respectively. The present study examines the oxidation of dimethylamino-cinnamaldehyde (DMAC), vanillin and phenanthridine by AO and xanthine by XOR from Drosophila cytosol. Therefore, the results obtained in the present study showed the DMAC, vanillin and phenanthridine substrates used were found to be good substrates of Drosophila AO and xanthine is the preferred substrate for Drosophila XOR. Km values of AO substrates were observed with DMAC (50±5.4 µM), phenanthridine (80±9.1 µM) and vanillin (303±11.7 µM) respectively for Drosophila cytosol. The Km values for DMAC and phenanthridine were ~6 and ~4 fold lower than that for vanillin as a substrate. The Km for XOR with xanthine using NAD+ as an electron acceptor was 27±4.1 µM. Relatively low Vmax values were obtained with phenanthridine (1.78±0.38 nmol/min/mg protein) and DMAC (1.80±0.35 nmol/min/mg protein). The highest Vmax was obtained from Drosophila cytosol with vanillin (7.58±2.11 nmol/min/mg protein). It is concluded these results that AO and XOR in Drosophila were able to catalyse the biotransformation of numerous substrates of the well-characterised mammalian AO and XOR. The kinetic parameters have indicated that the activity of AO of Drosophila may be a significant factor the oxidation of aromatic aldehyde compounds.

Keywords: aldehyde oxidase, xanthine oxidoreductase, dimethylamino-cinnamaldehyde, vanillin, phenanthridine, Drosophila melanogaster

Procedia PDF Downloads 404

Authors: Yijun Lai, Saber Khederzadeh, Lingshaung Han

Abstract:

Species in Thunnus are organized due to the similarity between them. The closeness between T. maccoyii, T. thynnus, T. Tonggol, T. atlanticus, T. albacares, T. obsesus, T. alalunga, and T. orientails are in different degrees. However, the genetic pattern of differentiation has not been presented based on individuals yet, to the author’s best knowledge. Hence, we aimed to analyze the difference in individuals level of tuna species to identify the factors that contribute to the maternal lineage variety using Cytochrome c oxidase subunit I (COXI) gene sequences. Our analyses provided evidence of sharing lineages in the Thunnus. A phylogenetic analysis revealed that these lineages are basal to the other sequences. We also showed a close connection between the T. tonggol, T. thynnus, and T. albacares populations. Also, the majority of the T. orientalis samples were clustered with the T. alalunga and, then, T. atlanticus populations. Phylogenetic trees and migration modeling revealed high proximity of T. thynnus sequences to a few T. orientalis and suggested possible gene flow with T. tonggol and T. albacares lineages, while all T. obsesus samples indicated unique clustering with each other. Our results support the presence of old maternal lineages in Thunnus, as a legacy of an ancient wave of colonization or migration.

Keywords: Thunnus Tuna, phylogeny, maternal lineage, COXI gene

Procedia PDF Downloads 246

Authors: V. Surena, B. Nazemisalman, F. Noghrehkar

Abstract:

Introduction: Gingival overgrowth (GO) is a common side effect involving users of antiepileptic, immunosuppressive and calcium channel blocker drugs. Cyclosporine and phenytoin are amongst the most widely used drugs associated with GO. Gingival fibroblasts seem to have a significant role in the production of certain enzymes after administration of the drugs contributing to GO. Previous studies have shown a higher prevalence of GO in children and adolescents. The aim of this study was to compare normal human gingival fibroblasts with those exposed to Cyclosporine or phenytoin in measuring the production levels of certain enzymes that could have a possible role in GO. Methods: samples were obtained from the gingival biopsies of seven adult and seven children and were cultured into plates. With the growth of fibroblast cells, they were treated with or without either Cyclosporine or phenytoin. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to determine the expressed levels of R-EGF, cathepsin B,L, Lysyl oxidase, COL1, TGF β1, MMP-1,2, and TIMP1. Results: according to RT-PCR analyses, the expressed levels of R-EGF, cathepsin B, L, Lysyl oxidase, COL1, TGF β1, MMP-1, 2 and TIMP1 were affected by Cyclosporine and phenytoin. TGF-β1, TIMP, Cathepsin B and EGF showed comparable values in the adult and pediatric groups. Conclusions: Different expressed levels of enzymes after treatment of the gingival fibroblasts of adults and pediatrics with phenytoin or Cyclosporine could be the reason for the higher severity of GO in children. More studies need to be performed on the pathogenesis of GO at different age groups.

Keywords: cyclosporine, fibroblasts, phenytoin, gingivae

Procedia PDF Downloads 235

Authors: Ofonime U. M. John, Justina I. R. Udotong, Victor O. Nwaugo, Ime R. Udotong

Abstract:

Oily wastes from oil and gas exploration and production (O&G E&P) activities were remediated for twelve weeks using Soil-Poultry dropping amendment. Culture-dependent microbiological, chemical and enzymatic techniques were employed to assess the efficacy of remediation process. Microbiological activities of the remediated wastes showed increased hydrocarbonoclastic microbial populations with increased remediation time; 2.7±0.1 x 105cfu/g to 8.3 ± 0.04 x106cfu/g for hydrocarbon utilizing bacteria, 1.7 ± 0.2 x103cfu/g to 6.0 ± 0.01 x 104cfu/g for hydrocarbon utilizing fungi and 2.2 ± 0.1 x 102cfu/g to 6.7 ± 0.1 x 103cfu/g for hydrocarbon utilizing actinomycetes. Bacteria associated with the remediated wastes after the remediation period included the genera Bacillus, Psuedomonas, Beijerinckia, Acinetobacter, Alcaligenes and Serratia. Fungal isolates included species of Penicillium, Aspergillus and Cladosporium, while the Actinomycetes included species of Rhodococcus, Nocardia and Streptomyces. Slight fluctuations in pH values between 6.5± 0.2 and 7.1 ± 0.08 were recorded throughout the process, while total petroleum hydrocarbon (TPH) content decreased from 89, 900 ± 0.03mg/kg to 425 ± 0.1 mg/kg after twelve weeks of remediation. The polycyclic aromatic hydrocarbon (PAH) levels decreased with increased remediation time; naphthalene, flourene, pheneanthrene, anthracene, pyrene, chrysene and benzo(b)flouranthene showed decreased values < 0.01 after twelve weeks of remediation. Enzyme activities revealed increased dehydrogenase and urease activities with increased remediation time and decreased phenol oxidase activity with increased remediation period. There was a positive linear correlation between densities of hydrocarbonoclastic microbes and dehydrogenase activity. On the contrary, phenol oxidase and urease activities showed negative correlation with microbial population. Results of this study confirmed that remediation of oily wastes using soil-poultry dropping amendment can result in eco-friendly O&G E&P wastes. It also indicates that urease and phenol oxidase activities can be reliable indices/tools to monitor PAH levels and rates of petroleum hydrocarbon degradation.

Keywords: dehydrogenase activity, oily wastes, remediation, soil-poultry dropping amendment

Procedia PDF Downloads 283

Authors: Gourav Jain, Sameer Dixit, Surjeet Kumar Arya, Praveen C. Verma

Abstract:

Plant cell wall plays a major role in defence mechanism against biotic and abiotic stress as it constitutes the physical barrier between the microenvironment and internal component of the cell. It is a complex structure composed of mostly carbohydrates among which cellulose and hemicelluloses are most abundant that is embedded in a matrix of pectins and proteins. Multiple enzymes have been reported which plays a vital role in cell wall modification, Pectin Methylesterase (PME) is one of them which catalyses the demethylesterification of homogalacturonans component of pectin which releases acidic pectin and methanol. As emitted methanol is toxic to the insect pest, we use PME gene for the better methanol production. In the current study we showed overexpression of PME gene isolated from Withania somnifera under the insect inducible promoter causes enhancement of methanol production at the time of insect feeds to plants, and that provides better insect resistance property. We found that the 85-90% mortality causes by transgenic tobacco in both chewing (Spodoptera litura larvae and Helicoverpa armigera) and sap-sucking (Aphid, mealybug, and whitefly) pest. The methanol content and emission level were also enhanced by 10-15 folds at different inducible time point interval (15min, 30min, 45min, 60min) which would be analysed by Purpald/Alcohol Oxidase method.

Keywords: methanol, Pectin methylesterase, inducible promoters, Purpald/Alcohol oxidase

Procedia PDF Downloads 205

Authors: Mutalubi Aremu Akintunde, John Isa

Abstract:

The negative impact on the atmosphere, of chlorofluorocarbon refrigerants (CFC) radical changes and measures were put in place to replace them. This has led to search for alternative refrigerants over the past decades. This paper presents thermal conductivity, diffusivity and performance of two alternative refrigerants as replacement to R12, which has been a versatile refrigerant which had turned the refrigeration industries around for decades, but one of the offensive refrigerants. The new refrigerants were coded RA1 (50%R600a/50%R134a;) and RA2 (70%R600a/30%R134a). The diffusivities for RA1 and RA2 were estimated to be, 2.76384 X 10-8 m2/s and 2.74386 X 10-8 m2/s respectively, while that of R12 under the same experimental condition is 2.43772 X 10-8 m2/s. The performances of the two refrigerants in a refrigerator initially designed for R12, were very close to that of R12. Other thermodynamic parameters showed that R12 can be replaced with both RA1 and RA2.

Keywords: alternative refrigerants, conductivity, diffusivity, performance, refrigerants

Procedia PDF Downloads 120