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Sample records for musca domestica acetylcholinesterase

  1. Variation of Musca domestica L. acetylcholinesterase in Danish housefly populations

    DEFF Research Database (Denmark)

    Kristensen, Michael; Huang, Jing; Qiao, Chuan-Ling

    2006-01-01

    Anti-cholinesterase resistance is in many cases caused by modified acetylcholinesterase (MACE). A comparison was made of toxicological data and AChE activity gathered from 21 field populations and nine laboratory strains of houseflies, Musca domestica L., to elucidate the best way of generating...... data to provide advice for management strategies and gathering information for resistance risk assessment on the organophosphates azamethiphos and dimethoate and the carbamate methomyl, which have been the primary insecticides used against adult houseflies in Denmark. Cluster analysis was performed...... and > 2000 houseflies were assigned to one of three phenotypes based on total acetylcholinesterase activity as well as inhibition by azamethiphos, methomyl or omethoate. A cluster, i.e. a phenotype, with high total AChE activity and high sensitivity to azamethiphos and less sensitivity to inhibition...

  2. Identification and characterization of mutations in housefly (Musca domestica) acetylcholinesterase involved in insecticide resistance

    DEFF Research Database (Denmark)

    Walsh, Sinead B.; Dolden, Tracey A.; Moores, Graham D.

    2001-01-01

    Acetylcholinesterase (AChE) insensitive to organophosphate and carbamate insecticides has been identified as a major resistance mechanism in numerous arthropod species. However, the associated genetic changes have been reported in the AChE genes from only three insect species; their role in confe...... of the AChE protein from Torpedo californica and D. melanogaster....

  3. Pertumbuhan dan Reproduksi Lalat Musca domestica pada Berbagai Media Perkembangbiakan

    Directory of Open Access Journals (Sweden)

    Endang Puji Astuti

    2013-02-01

    Full Text Available Abstract. Musca domestica was insect that disturbing human and other animal. Intensity M. Domestica with human living includes public healthy. The aim of research is to observe succeed of Mdomestica in breeding wether difJerent place. Larva growth in each stadia was different, 2-4 days average. Its contributed by humidity. On dry container, pupae that succes eksklosi only 50%. Presentation offood show difJerences oflarvae development. 3rd stage of larvae that has enough food is 17mm in length and 12mm in length for larvae with less food. Salty water make larvae growing faster than water without salto Development of Musca domestica in laboratory injluenced by high humidity, preset of food and adaptation of Musca domestica its self.Keywords : Musca domestica, rearing, development, humidity, food, salt - water

  4. Antimicrobial activity of alcohols from Musca domestica.

    Science.gov (United States)

    Gołębiowski, Marek; Dawgul, Małgorzata; Kamysz, Wojciech; Boguś, Mieczysława I; Wieloch, Wioletta; Włóka, Emilia; Paszkiewicz, Monika; Przybysz, Elżbieta; Stepnowski, Piotr

    2012-10-01

    Information on the stimulatory and inhibitory effects of cuticular alcohols on growth and virulence of insecticidal fungi is unavailable. Therefore, we set out to describe the content of cuticular and internal alcohols in the body of housefly larvae, pupae, males and females. The total cuticular alcohols in larvae, males and females of Musca domestica were detected in comparable amounts (4.59, 3.95 and 4.03 μg g(-1) insect body, respectively), but occurred in smaller quantities in pupae (2.16 μg g(-1)). The major free alcohol in M. domestica larvae was C(12:0) (70.4%). Internal alcohols of M. domestica larvae were not found. Among cuticular pupae alcohols, C(12:0) (31.0%) was the most abundant. In the internal lipids of pupae, only five alcohols were identified in trace amounts. The most abundant alcohol in males was C(24:0) (57.5%). The percentage content of cuticular C(24:0) in males and females (57.5 and 36.5%, respectively) was significantly higher than that of cuticular lipids in larvae and pupae (0.9 and 5.6%, respectively). Only two alcohols were present in the internal lipids of males in trace amounts (C(18:0) and C(20:0)). The most abundant cuticular alcohols in females were C(24:0) (36.5%) and C(12:0) (26.8%); only two alcohols (C(18:0) and C(20:0)) were detected in comparable amounts in internal lipids (3.61±0.32 and 5.01±0.42 μg g(-1), respectively). For isolated alcohols, antimicrobial activity against 10 reference strains of bacteria and fungi was determined. Individual alcohols showed approximately equal activity against fungal strains. C(14:0) was effective against gram-positive bacteria, whereas gram-negative bacteria were resistant to all tested alcohols. Mixtures of alcohols found in cuticular lipids of larvae, pupae, males and females of M. domestica generally presented higher antimicrobial activity than individual alcohols. In contrast, crude extracts containing both cuticular and internal lipids showed no antifungal activity against the

  5. Circadian rhytm in the house fly (Musca domestica)

    OpenAIRE

    BAZALOVÁ, Olga

    2010-01-01

    In this thesis the expression pattern of five circadian clock genes (cwo, pdp 1{$\\varepsilon$}, ck 1{$\\varepsilon$}, ck 2{$\\beta$} and pdh) was studied in the housefly (Musca domestica). The influence of temperature on the expression pattern of these five genes and of two others genes, per and tim, was examined. The locomotor activity of flies exposed to three different temperature conditions was studied.

  6. Wound Myiasis Due to Musca domestica (Diptera: Muscidae in Persian Horned Viper, Pseudocerastes persicus (Squamata: Viperidae

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    R Dehghani

    2012-06-01

    Full Text Available A case of myiasis due to Musca domestica describes in Pseudocerastes persicus for the first time. The snake was found in Bari Karafs, Kashan, Iran, with a lesion on its body. Fourteen live larvae of M. domestica removed from its wound. This is the first report of a new larval habitat of M. domestica.

  7. Using the fungus Entomophthora muscae (chon Fresenius to eliminate some larval roles of Musca domestica

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    Walaa Yas Lahmood

    2017-07-01

    Full Text Available Studied effect serial concentrations from spores filtrate of fungus Entomophthora muscae on some larval roles of musca domestica in laboratory. Results were made clear that the insect roles are sensitive to fungus, and treated the food larva of musca domestica and sprinkle it by concentration 2.8×106 , 2.8×107, 2.8×108 (spore/ml has led to get rates of destruction of cumulative faculty certified on the concentration and time its magnitude 16.60 , 47.67, 53.30 % respectively , also recorded some phenotypic distortion infected dead larva represent by contraction and blackening body. The treatment of pupael by sprinkling the previous fungus concentration recorded rate of destruction of accumulative faculty its magnitude 13.33, 26.67, 33.33% respectively, also the rates emergence of adults ranged between 66.67 – 86.67 % in comparison with rates of emergence of adults in control treatment 96.67% The results are made clear that adults treatment by sprinkle with last concentration from fungus spore filtrate recorded rates of distraction its magnitude 46.61, 56.67, 70% respectively after one week from treatment .

  8. Musca domestica laboratory susceptibility to three ethnobotanical culinary plants.

    Science.gov (United States)

    El Zayyat, Elham A; Soliman, Mohammed I; Elleboudy, Noha A; Ofaa, Shaimaa E

    2015-10-01

    Throughout history, synanthropic Musca domestica had remained a worldwide problem whenever poor sanitation and bad hygienic conditions exists. Houseflies growing resistance to chemical insecticides are a rising environmental problem that necessitates search for alternatives. Mentha cervina, Ocimum basilicum, and Coriandrum sativum were tested for bioactivity on M. domestica adults and larvae. They are culinary Mediterranean plants. In adulticidal bioassay, using both CDC bottles and fumigation techniques, basil was the most effective extract with LC50 1.074 and 34.996 g/L, respectively. Concerning larvicidal bioassay by fumigation technique, coriander had the highest toxicity index with LC50 29.521 g/L. In both dipping and feeding technique, basil had the highest toxicity with LC50 32.643 and 0.749 g/L, respectively. Basil showed the highest toxicity results in four out of the five models tested followed by coriander then mint; this result highlights the potentiality of basil as a green insecticide in management of flies and opens new insight in the industrialization of basil-based fly control products.

  9. Evolution of resistance to pyrethroid insecticides in Musca domestica.

    Science.gov (United States)

    Scott, Jeffrey G

    2017-04-01

    Houseflies, Musca domestica L., are a significant pest because of the numerous diseases they transmit. Control of housefly populations, particularly at animal production facilities, is frequently done using pyrethroid insecticides which kill insects by prolonging the open time of the voltage-sensitive sodium channel (VSSC). Houseflies have evolved resistance to pyrethroids owing to mutations in Vssc and by cytochrome-P450-mediated detoxification. Three Vssc mutations are known: kdr (L1014F), kdr-his (L1014H) and super-kdr (M918T + L1014F). Generally, the levels of resistance conferred by these mutations are kdr-his resistance than kdr. P450-mediated resistance can result from overexpression of CYP6D1 or another P450 (unidentified) whose overexpression is linked to autosomes II or V. The initial use of field-stable pyrethroids resulted in different patterns of evolution across the globe, but with time these mutations have become more widespread in their distribution. What is known about the fitness costs of the resistance alleles in the absence of insecticide is discussed, particularly with respect to the current and future utility of pyrethroid insecticides. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  10. Adaptation of Musca domestica L. field population to laboratory breeding causes transcriptional alterations

    DEFF Research Database (Denmark)

    Højland, Dorte H.; Vagn Jensen, Karl-Martin; Kristensen, Michael

    2014-01-01

    The housefly, Musca domestica, has developed resistance to most insecticides applied for its control. Expression of genes coding for detoxification enzymes play a role in the response of the housefly when encountered by a xenobiotic. The highest level of constitutive gene expression of nine P450 ...

  11. Interaction of some organophosphorus compounds in susceptible and resistant houseflies (Musca domestica L.)

    NARCIS (Netherlands)

    Abdallah, M.D.

    1963-01-01

    The effect of tri-ortho-cresylphosphate (TOCP) on poisoning by parathion and paraoxon in susceptible and resistant houseflies (Musca domestica L.) was investigated.

    TOCP influenced the penetration of parathion and paraoxon, and this could explain the contradictory results of both an

  12. Temperature and Population Density Effects on Locomotor Activity of Musca domestica (Diptera: Muscidae)

    DEFF Research Database (Denmark)

    Schou, T. M.; Faurby, S.; Kjærsgaard, A.

    2013-01-01

    The behavior of ectotherm organisms is affected by both abiotic and biotic factors. However, a limited number of studies have investigated the synergistic effects on behavioral traits. This study examined the effect of temperature and density on locomotor activity of Musca domestica (L.). Locomot...

  13. The enhancement by caffeine of the frequency of lethal dominant mutation induced by gamma radiation in oocytes of Musca domestica

    International Nuclear Information System (INIS)

    Targa, H.J.; Rogatko, A.

    1982-01-01

    The results obtained, when a new technique for feeding insects is employed, on the effects of caffeine of the radiation - induced breaks of oocyte chromatids of Musca domestica are presented. (M.A.) [pt

  14. Melia azedarach L. extracts and their activity on Musca domestica L. (Diptera: Muscidae

    Directory of Open Access Journals (Sweden)

    Marise M. O. Cabral

    Full Text Available Crudes extracts and fractions from seeds of Melia azedarach L. (Meliaceae have been assayed on Musca domestica Linnaeus, 1758 (Diptera: Muscidae. Thus, the post-embryonic development of the flies was reduced and the delay from newly hatched larvae to adults had significant increase. In addition, the pupal weights were reduced and the sexual ratio altered. Toxicity to fly eggs was also observed.

  15. Survival and Movement of Insect Parasitic Nematodes in Poultry Manure and Their Infectivity Against Musca domestica

    OpenAIRE

    Georgis, Ramon; Mullens, Bradley A.; Meyer, Jeffery A.

    1987-01-01

    Survival, infectivity, and movement of three insect parasitic nematodes (Steinernema feltiae All strain, S. bibionis SN strain, and Heterorhabditis heliothidis NC strain) in poultry manure were tested under laboratory conditions. The majority (70-100%) of the nematodes died within 18 hours after exposure to the manure. Nematodes exposed to manure slurry for 6 hours killed at least 95% of the house fly larvae, Musca domestica, but nematodes exposed for 12 hours achieved less than 40% larval mo...

  16. Fatal attraction. Control of the housefly (Musca domestica)

    NARCIS (Netherlands)

    Smallegange, R.C.

    2004-01-01

    Despite its long association with man, the housefly (Musea domestica Linnaeus) remains one of the most difficult pests to control. It is a ubiquitous insect that can be found in houses, stables, food processing factories and other domesticated areas and buildings. Reliance on insecticides for fly

  17. Effects of Persea americana Mill. seed extracts on the postembryonic development of Musca domestica (Diptera: Muscoide

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    Silvia del C. Molina Bertrán

    2018-04-01

    Full Text Available Context: The synthetic insecticides used to control Diptera are harmful to the environment and humans. Extracts and compounds from plants are a more sustainable source for the development of bio-insecticides. Aims: To evaluate the efficacy of a hydroalcoholic extract of Persea americana Mill seeds as an alternative control of the species Musca domestica. Methods: The extracts were obtained by two methods, the Shaker (S and the Soxhlet extraction (SE method, using 94% ethanol as the solvent. Also, the qualitative chemical composition was determined by phytochemical screening. The effect of the two extracts on the post-embryonic development of the fly as well as the adulticidal effect was evaluated. Results: Phytochemical analysis revealed the presence of metabolites such as alkaloids, coumarins, tannins, flavonoids, sugars and amino acids. The influence on the post-embryonic development of M. domestica was demonstrated, especially on the viability of larvae and neolarvae to adults; however, the effect on the weight and duration of each period was low. The adulticidal effects of the extracts were determined by the lethal concentration 50(LC50 of 2.910 mg/100 mL and 3.944 mg/100 mL for the S and SE extracts, respectively. Conclusions: Both extracts showed their insecticidal effects against Musca domestica, but the extract elaborated by S method showed greater influence diminishing viability and better adulticidal effect.

  18. Larvicidal potential of essential oils against Musca domestica and Anopheles stephensi.

    Science.gov (United States)

    Chauhan, Nitin; Malik, Anushree; Sharma, Satyawati; Dhiman, R C

    2016-06-01

    The larvicidal activity of Mentha piperita, Cymbopogan citratus (lemongrass), Eucalyptus globulus and Citrus sinensis (orange) essential oils and their combinations was evaluated against Musca domestica (housefly) and Anopheles stephensi (mosquitoes) through contact toxicity assay. Among all the tested essential oils/combinations, Me. piperita was found to be the most effective larvicidal agent against Mu. domestica and An. stephensi with LC50 values of 0.66 μl/cm(2) and 44.66 ppm, respectively, after 48 h. The results clearly highlighted that the addition of mentha oil to other oils (1:1 ratio) improved their larvicidal activity. The order of effectiveness of essential oils/combinations indicated that the pattern for An. stephensi follows the trend as mentha > mentha + lemongrass > lemongrass > mentha + eucalyptus > eucalyptus > mentha + orange > orange and for Mu. domestica as mentha > mentha + lemongrass > lemongrass > mentha + orange > orange > mentha + eucalyptus > eucalyptus. The images obtained from scanning electron microscopy (SEM) analysis indicated the toxic effect of Me. piperita as the treated larvae were observed to be dehydrated and deformed. This study demonstrates the effectiveness of tested essential oils/combinations against the larval stages of Mu. domestica and An. stephensi and has the potential for development of botanical formulations.

  19. Effect of Triflumuron and Pyriproxyfen on Musca domestica L Larval Stages in the Laboratory

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    S Sulaiman

    2008-08-01

    Full Text Available Background:  The residual effect of triflumuron and pyriproxyfen on Musca domestica L larval stages was studied in the laboratory."n"nMethods: Both IGRs at varying concentrations ranging between 0.5 to 2.5 mg/L were placed inside beakers with mice chow and vitamin B complex and water as food for the 1st, 2nd and 3rd instars M.domestica larvae."n"nResults: Both IGRs inhibit M. domestica adult emergence of 98-98.5% when applied at the lowest concentration of 0.5 mg/L on the 1st instar, 93-97% of adult emergence inhibition on the 2nd instar,and 91-97% of adult emergence inhibi­tion on the 3rd instar larvae respectively. There was no significant difference between triflumuron and pyriproxy­fen on house­fly adult emergence inhibition when fed to the 1st, 2nd and 3rd instars M.domestica larvae (P> 0.05. However, there was a significant difference between the IGRs and the control (P< 0.05."n"nConclusion: Both triflumuron and pyriproxyfen are effective in inhibiting adult emergence of housefly M  domes­tica and therefore should be recommended for fly control particularly in chicken farms and dumping grounds in Malaysia for housefly control activities.

  20. Effect of four commercial fungal formulations on mortality and sporulation of house flies (Musca domestica) and stable flies (Stomoxys calcitrans)

    Science.gov (United States)

    House flies (Musca domestica L.) and stable flies (Stomoxys calcitrans (L.)) (Diptera: Muscidae) are major pests of livestock. Biological control is an important tool in an integrated control framework. Increased mortality in filth flies has been documented with entomopathogenic fungi, and several s...

  1. Effects of relative humidity, temperature, and population density on production of cuticular hydrocarbons in housefly Musca domestica L.

    NARCIS (Netherlands)

    Noorman, N; Den Otter, CJ

    The production of cuticular hydrocarbons by both males and females of Musca domestica L. under very wet conditions (90% relative humidity) compared to the production at 50 and 20% relative humidity is delayed up to at least 3 days after emergence from the pupae. Eight days after emergence, however,

  2. Genome of the house fly, Musca domestica L., a global vector of diseases with adaptations to a septic environment

    NARCIS (Netherlands)

    Scott, Jeffrey G; Warren, Wesley C; Beukeboom, Leo W; Bopp, Daniel; Clark, Andrew G; Giers, Sarah D; Hediger, Monika; Jones, Andrew K; Kasai, Shinji; Leichter, Cheryl A; Li, Ming; Meisel, Richard P; Minx, Patrick; Murphy, Terence D; Nelson, David R; Reid, William R; Rinkevich, Frank D; Robertson, Hugh M; Sackton, Timothy B; Sattelle, David B; Thibaud-Nissen, Francoise; Tomlinson, Chad; van de Zande, Louis; Walden, Kimberly; Wilson, Richard K; Liu, Nannan

    2014-01-01

    BACKGROUND: Adult house flies, Musca domestica L., are mechanical vectors of more than 100 devastating diseases that have severe consequences for human and animal health. House fly larvae play a vital role as decomposers of animal wastes, and thus live in intimate association with many animal

  3. INFECTIVITY OF TWO MEMBERS OF THE ENTOMOPHTHORA MUSCAE COMPLEX [ZYGOMYCETES: ENTOMOPHTHORALES] FOR MUSCA DOMESTICA [DIPT.: MUSCIDAE

    DEFF Research Database (Denmark)

    Bellini, R; Mullens, B A; Jespersen, J B

    1992-01-01

    Dose-mortality studies were conducted with 2 members of the Entomophthora muscae (Cohn) Fresenius complex from southern California (CA) and Denmark (DA) infecting house flies, Musca domestica L., from southern California. Primary conidia of the DA form were significantly more infective (LC50 = 34...

  4. Molecular detection and antimicrobial resistance of Aeromonas from houseflies (Musca domestica in Iran

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    Davood Ommi

    2015-11-01

    Full Text Available Objective. This study aimed to report the molecular detection and antimicrobial resistance of Aeromonas among houseflies (Musca domestica in Shahrekord and Isfahan provinces of Iran. Materials and methods. Flies were caught from household kitchens, cattle farms, animal hospitals, human hospitals, slaughter house and poultry farms and put in collection separate sterile tubes. Isolation was accomplished by culture of flies in alkaline peptone water followed by identification with Aeromonas-specific Polymerase Chain Reaction (PCR. Results. Out of 600 houseflies 73 (12.2% were infected with Aeromonas spp. Significantly higher frequencies of Aeromonas were isolated in Shahrekord province (13.0%; 39/300 than in Isfahan province (11.3%; 34/300. The recovery frequencies of the organisms were significantly lower in kitchens as compared to those in cattle farms and hospital wards which were similar. Higher proportions of infected flies were obtained during summer whereas low proportions were obtained during winter. Conclusions. It is concluded that houseflies do harbor diarrheagenic pathogens, including Aeromonas especially during summer. The carried organisms are resistant to a number of antimicrobials at different levels. Thus, future plans aimed at stemming infections caused by these organisms should take flies into account. Control efforts of infections caused by this particular bacterium should therefore take into account Musca domestica.

  5. UJI EFIKASI LARVASIDA BERBAHAN AKTIF DICHLORVOS TERHADAP LARVA LALAT RUMAH Musca domestica DI LABORATORIUM

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    Hadi Suwasono

    2012-10-01

    Full Text Available The house fly, Musca domestica have been implicated in the mechanical transmission of pathogens causing diseases. Important diseases that may be transmitted under certain conditions include infection of virus, bacterial, protozoan, fungal and helminthic. Therefore control measures should be carried out only when the presence of the house fly is intolerable to humans due to their being a health hazard or nuisance. The larvicide with active ingredient of dichlorvos 250 g/l was applied to the house fly larvae which conducted in the laboratory. There were jive concentrations of larvicides to be tested i.e. 6; 12; 24; 48 and 96 ml by product/ 3 l of water/m2. A dried powder of chicken dung was utilized as larvae medium. The medium was inoculated with the third instar larvae then it sprayed with the larvicide suspension. All larvicidal assays were carried out at 26 - 30°C temperature and 60-80% RH in the laboratory. The results revealed that all of the five tested concentrations yielded between 0 and 5.2 percent larval mortalities.   Keywords: house fly, Musca domestica, larvicide, dichlorvos

  6. Evaluation of the house fly Musca domestica as a mechanical vector for an anthrax.

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    Antonio Fasanella

    Full Text Available Anthrax is a disease of human beings and animals caused by the encapsulated, spore-forming, Bacillus anthracis. The potential role of insects in the spread of B. anthracis to humans and domestic animals during an anthrax outbreak has been confirmed by many studies. Among insect vectors, the house fly Musca domestica is considered a potential agent for disease transmission. In this study, laboratory-bred specimens of Musca domestica were infected by feeding on anthrax-infected rabbit carcass or anthrax contaminated blood, and the presence of anthrax spores in their spots (faeces and vomitus was microbiologically monitored. It was also evaluated if the anthrax spores were able to germinate and replicate in the gut content of insects. These results confirmed the role of insects in spreading anthrax infection. This role, although not major, given the huge size of fly populations often associated with anthrax epidemics in domestic animals, cannot be neglected from an epidemiological point of view and suggest that fly control should be considered as part of anthrax control programs.

  7. Plasticity in behavioural responses and resistance to temperature stress in Musca domestica

    DEFF Research Database (Denmark)

    Kjaersgaard, Anders; Blackenhorn, Wolf U.; Pertoldi, Cino

    2015-01-01

    , at the stressful high temperature Spanish flies flew the furthest and Danish flies the shortest distance. Neither body size nor wing loading affected flight performance, although flies with narrower wings tended to fly further (wing shape effect). Swiss flies were most active in terms of locomotor activity......Organisms can respond to and cope with stressful environments in a number of ways including behavioural, morphological and physiological adjustments. To understand the role of behavioural traits in thermal adaptations we compared heat resistance, locomotor (walking and flying) activity, flight...... performance and morphology of three European populations of Musca domestica (Diptera: Muscidae) originating from different thermal conditions (Spain, Switzerland and Denmark) at benign and stressful high temperatures. Spanish flies showed greater heat resistance than Swiss and Danish flies. Similarly...

  8. Ophthalmomyiasis externa by Musca domestica in a case of orbital metastasis

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    Rose Mary Tomy

    2013-01-01

    Full Text Available Infestation of the living tissues of the eye, ocular adnexae or orbit by larvae of flies of the order Diptera is known as ophthalmomyiasis. Ophthalmomyiasis externa is a limited superficial infestation of external ocular structures such as conjunctiva and adnexae. A case of follicular carcinoma thyroid with orbital metastasis presenting as ophthalmomyiasis externa is reported. The patient presented with foul smelling ulcers of the medial and lateral orbital walls of the right eye extending into the lids and nose, teeming with maggots. Computerized Tomography (CT showed widespread bone destruction with extension into surrounding sinuses. Over 200 maggots were manually removed after immobilisation with turpentine oil instillation. Entomological examination showed it to be a case of accidental myiasis caused by the common housefly Musca domestica. The patient was managed conservatively and the ulcer cavities filled up with healthy granulation tissue. This case is presented on account of its rarity.

  9. Biologically based machine vision: signal analysis of monopolar cells in the visual system of Musca domestica.

    Science.gov (United States)

    Newton, Jenny; Barrett, Steven F; Wilcox, Michael J; Popp, Stephanie

    2002-01-01

    Machine vision for navigational purposes is a rapidly growing field. Many abilities such as object recognition and target tracking rely on vision. Autonomous vehicles must be able to navigate in dynamic enviroments and simultaneously locate a target position. Traditional machine vision often fails to react in real time because of large computational requirements whereas the fly achieves complex orientation and navigation with a relatively small and simple brain. Understanding how the fly extracts visual information and how neurons encode and process information could lead us to a new approach for machine vision applications. Photoreceptors in the Musca domestica eye that share the same spatial information converge into a structure called the cartridge. The cartridge consists of the photoreceptor axon terminals and monopolar cells L1, L2, and L4. It is thought that L1 and L2 cells encode edge related information relative to a single cartridge. These cells are thought to be equivalent to vertebrate bipolar cells, producing contrast enhancement and reduction of information sent to L4. Monopolar cell L4 is thought to perform image segmentation on the information input from L1 and L2 and also enhance edge detection. A mesh of interconnected L4's would correlate the output from L1 and L2 cells of adjacent cartridges and provide a parallel network for segmenting an object's edges. The focus of this research is to excite photoreceptors of the common housefly, Musca domestica, with different visual patterns. The electrical response of monopolar cells L1, L2, and L4 will be recorded using intracellular recording techniques. Signal analysis will determine the neurocircuitry to detect and segment images.

  10. The Effects of Sex-Ratio and Density on Locomotor Activity in the House Fly, Musca domestica

    OpenAIRE

    Bahrndorff, Simon; Kjærsgaard, Anders; Pertoldi, Cino; Loeschcke, Volker; Schou, Toke M.; Skovgård, Henrik; Hald, Birthe

    2012-01-01

    Although locomotor activity is involved in almost all behavioral traits, there is a lack of knowledge on what factors affect it. This study examined the effects of sex-ratio and density on the circadian rhythm of locomotor activity of adult Musca domestica L. (Diptera: Muscidae) using an infra-red light system. Sex-ratio significantly affected locomotor activity, increasing with the percentage of males in the vials. In accordance with other studies, males were more active than females, but th...

  11. Crystallization, data collection and phasing of two digestive lysozymes from Musca domestica

    Energy Technology Data Exchange (ETDEWEB)

    Marana, S. R.; Cançado, F. C. [Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo (Brazil); Valério, A. A. [Centro de Biologia Molecular e Estrutural (CeBiMe), Laboratório Nacional de Luz Síncrotron (LNLS), CP 6192, Campinas, SP 13084-971 (Brazil); Departamento de Bioquímica, Universidade Estadual de Campinas, Campinas (Brazil); Ferreira, C.; Terra, W. R. [Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo (Brazil); Barbosa, J. A. R. G., E-mail: joao@lnls.br [Centro de Biologia Molecular e Estrutural (CeBiMe), Laboratório Nacional de Luz Síncrotron (LNLS), CP 6192, Campinas, SP 13084-971 (Brazil); Departamento de Bioquímica, Universidade Estadual de Campinas, Campinas (Brazil); Departamento de Genética e Evolução, Universidade Estadual de Campinas, Campinas (Brazil); Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo (Brazil)

    2006-08-01

    The digestive lysozymes 1 and 2 from M. domestica were crystallized by vapour diffusion. The crystallographic data were processed to a maximum resolution of 1.9 Å in both cases. Lysozymes are mostly known for their defensive role against bacteria, but in several animals lysozymes have a digestive function. Here, the initial crystallographic characterization of two digestive lysozymes from Musca domestica are presented. The proteins were crystallized using the sitting-drop vapour-diffusion method in the presence of ammonium sulfate or PEG/2-propanol as the precipitant. X-ray diffraction data were collected to a maximum resolution of 1.9 Å using synchrotron radiation. The lysozyme 1 and 2 crystals belong to the monoclinic space group P2{sub 1} (unit-cell parameters a = 36.52, b = 79.44, c = 45.20 Å, β = 102.97°) and the orthorhombic space group P2{sub 1}2{sub 1}2 (unit-cell parameters a = 73.90, b = 96.40, c = 33.27 Å), respectively. The crystal structures were solved by molecular replacement and structure refinement is in progress.

  12. Efficacy of Essential Oils from Edible Plants as Insecticides Against the House Fly, Musca Domestica L.

    Directory of Open Access Journals (Sweden)

    Sara M. Palacios

    2009-05-01

    Full Text Available The compositions of 12 essential oils (EOs obtained by hydrodistillation of edible fruits and herbs were analyzed by gas chromatography/mass spectroscopy (GC/MS. The insecticidal activity of each oil against the house fly Musca domestica was evaluated by placing flies in a glass jar with a screw cap that held a piece of EO-treated cotton yarn. The dose necessary to kill 50% of flies (LC50 in 30 min was determined at 26 ± 1°C. Twelve EOs and 17 individual terpenes were assayed against M. domestica, showing LC50 values ranging from 3.9 to 85.2 and from 3.3 to >100 mg/dm3, respectively. EO from Citrus sinensis was the most potent insecticide (LC50 = 3.9 mg/dm3, followed by EOs from C. aurantium (LC50 = 4.8 mg/dm3 and Eucalyptus cinerea (LC50 = 5.5 mg/dm3. According to GC/MS analysis, limonene (92.47%, linalool (1.43%, and b-myrcene (0.88% were the principal components of C. sinensis EO. Limonene was also the principal constituent (94.07% of C. aurantium, while 1,8-cineole (56.86% was the major constituent of E. cinerea EO. 1,8-Cineole was most active against M. domestica (LC50 = 3.3 mg/dm3, while (4R(+-limonene, was moderately active (LC50 = 6.2 mg/dm3. Dimethyl 2,2-dichlorovinyl phosphate (DDVP selected as a positive control, showed an LC50 of 0.5 mg/dm3. EOs from C. sinensis, C. aurantium, and E. cinerea show promise as natural insecticides against houseflies.

  13. Differential distribution of glutamate- and GABA-gated chloride channels in the housefly Musca domestica.

    Science.gov (United States)

    Kita, Tomo; Ozoe, Fumiyo; Azuma, Masaaki; Ozoe, Yoshihisa

    2013-09-01

    l-Glutamic acid (glutamate) mediates fast inhibitory neurotransmission by affecting glutamate-gated chloride channels (GluCls) in invertebrates. The molecular function and pharmacological properties of GluCls have been well studied, but not much is known about their physiological role and localization in the insect body. The distribution of GluCls in the housefly (Musca domestica L.) was thus compared with the distribution of γ-aminobutyric acid (GABA)-gated chloride channels (GABACls). Quantitative PCR and ligand-binding experiments indicate that the GluCl and GABACl transcripts and proteins are predominantly expressed in the adult head. Intense GluCl immunostaining was detected in the lamina, leg motor neurons, and legs of adult houseflies. The GABACl (Rdl) immunostaining was more widely distributed, and was found in the medulla, lobula, lobula plate, mushroom body, antennal lobe, and ellipsoid body. The present findings suggest that GluCls have physiological roles in different tissues than GABACls. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Gamma-irradiation of pupae of the house fly musca domestica L., and adult survival

    International Nuclear Information System (INIS)

    Guneidy, A.M.; Abdu, R.M.; Hamed, M.S.

    1975-01-01

    Effects of gamma-irradiation on pupae of the house fly, Musca domestica L., were determined. Pupae of 1, 2, 3, 4 and 5-day old, when exposed to a single dose of gamma rays, showed that the younger pupae were more susceptible than older pupae. The 1- and 2-day age groups were fully susceptible to all levels of treatment where no emergence was observed. The 3-day and 4-day age groups were of equal susceptibility to irradiation where emergence of 23.50% and 24.84%, respectively, was recorded at 5 Kr only. Pupae failed to emerge at dosages of 10, 15 and 25 Kr. Emergence was observed at all dosages in the 5-day old group of pupae where there was a correlation between dosage and emergence. At 5 Kr and 10 Kr, 85% and 66.36% of flies, respectively, emerged. This figure declined to 22.54% at 15 Kr and 4.09% at 25 Kr. Males were less susceptible than females. Flies of both sexes emerging from irradiated pupae died earlier than the control flies

  15. Effiacy of citronella and eucalyptus oils against Musca domestica, Cimex lectularius and Pediculus humanus

    Directory of Open Access Journals (Sweden)

    Jehangir Khan

    2017-11-01

    Full Text Available Objective: To assess potential impacts of two indigenous plant oils: the citronella (Cymbopogon nardus and eucalyptus (Eucalyptus globulus for their insecticidal effect against Musca domestica (house fly, Cimex lectularius (bed bug and Pediculus humanus (louse. Methods: The oils of these two medicinal plants were applied separately at various concentrations (1, 2 and 3 mL/cup in cups lined with filter paper containing the target insects. Mortality was evaluated after fixed intervals (6, 12 and 24 h subsequent to the release of adult insects. Results: Results showed that both oils exhibited concentration and time dependent mortality against the tested insects. Data pertaining to present investigation clearly showed that percentage mortality owing to these botanicals against these medical pests was significantly high (98.33% at the rate of 3 mL for 24 h of exposure, followed by 2 and 1 mL concentrations with 12 and 6 h of exposure times. Conclusions: The results suggest that these plant oils possess good insecticidal properties against house fly, bed bug, and louse, and are safe to humans. Furthermore, the molecular (biochemical based study of these botanicals against diverse species of pests will be of much significance to control these pest insects.

  16. Pertumbuhan dan perkembangan larva Musca domestica Linnaeus (Diptera: Muscidae dalam beberapa jenis kotoran ternak

    Directory of Open Access Journals (Sweden)

    Ramadhani Eka Putra

    2015-09-01

    Full Text Available House fly (Musca domestica is an important urban insect that can transmit various infectious diseases. This insect usually utilized organic wastes as nutrition source for their larvae. One of the main sources of organic wastes is livestock manure produced by animal farming located near human dwelling area. Thus, appropriate house fly population control program at animal farm is needed,based on information on the house fly’s life history. The research is focused on the development of house fly larvae reared with different livestock manures, such as cow, chicken, and horse. As comparison, rice bran were used as control. Results showed that larvae reared with horse manure has the shortest development time (5 days, with lowest larval survival rate (30%, pupal weight (6.8 ± 0.141 g, and weight of female imago (4.9 ± 0.14 g. This finding indicates the lowest nutrition value of horse manure for house flies larvae. Further research is needed to find the effect of manure to variables that directly influence population growth, such as fecundity of female flies and egg survivorship. These additional information on life history will help the design of appropriate house fly population management program for animal farm.

  17. Development of Supputius cincticeps (Heteroptera, Pentatomidae) fed with Zophobas confusa, Tenebrio molitor (Coleoptera, Tenebrionidae) and Musca domestica (Diptera, Muscidae) larvae

    OpenAIRE

    Beserra, Eduardo B.; Zanuncio, Teresinha V.; Zanuncio, José C.; Santos, Germi P.

    1995-01-01

    Egg viability and nymphal development of the predatory bug Supputius cincticeps (Stål, 1860) were evaluated during two generations in the Biological Control Laboratory of the Núcleo de Biotecnologia Aplicada à Agropecuária (Bioagro/UFV) in Viçosa (Minas Gerais, Brazil) at 24.72±1.10ºC and photophase of 12 hours. Three treatments were represented by S. cincticeps fed with Zophobas confusa Gebien, 1906, Tenebrio molitor Linnaeus, 1758 and Musca domestica Linnaeus, 1758 larvae. Higher egg viabil...

  18. Search efficiency of Spalangia cameroni and Muscidifurax raptor on Musca domestica pupae in dairy cattle farms in Denmark

    DEFF Research Database (Denmark)

    Skovgård, Henrik

    2006-01-01

    significantly to overall parasitism, M. raptor had a significantly higher attack rate when first a female had located bags with sentinel pupae. Based on the above results, however, S. cameroni seems the most appropriate species for managing house flies in straw bedded dairy cattle farms in Denmark. A biological......Indoor releases of Spalangia cameroni Perkins and Muscidifurax raptor Girauelt & Sanders (Hymenoptera: Pteromalidae) were conducted in five organic dairy cattle farms to evaluate the overall effect on parasitism and efficiency at different pupal depths of Musca domestica L. (Diptera: Muscidae...... control strategy of simultaneous releases of S. cameroni and M. raptor is discussed. Key words: biological control, dairy cattle, dispersion, parasitism, Phygadeuon fumat...

  19. The effects of sex-ratio and density on locomotor activity in the house fly, Musca domestica

    DEFF Research Database (Denmark)

    Bahrndorff, Simon; Kjaersgaard, Anders; Pertoldi, Cino

    2012-01-01

    Although locomotor activity is involved in almost all behavioral traits, there is a lack of knowledge on what factors affect it. This study examined the effects of sex-ratio and density on the circadian rhythm of locomotor activity of adult Musca domestica L. (Diptera: Muscidae) using an infra......-red light system. Sex-ratio significantly affected locomotor activity, increasing with the percentage of males in the vials. In accordance with other studies, males were more active than females, but the circadian rhythm of the two sexes was not constant over time and changed during the light period...... of the behavioral interactions between houseflies and highlight the importance of these factors when designing behavioral experiments using M. domestica....

  20. Evaluation of Beauveria bassiana infection in the hemolymph serum proteins of the housefly, Musca domestica L. (Diptera: Muscidae).

    Science.gov (United States)

    Mishra, Sapna; Kumar, Peeyush; Malik, Anushree

    2017-11-01

    Beauveria bassiana plays a prominent role in biocontrol of houseflies, Musca domestica (L.). Thus, a deeper insight into immune response of M. domestica during B. bassiana infection was warranted to assist the production of more efficient mycoinsecticides. The present study investigates changes in protein profile of M. domestica hemolymph serum post B. bassiana infection using two-dimensional difference gel electrophoresis (2D-DIGE) followed by identification of selected proteins by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). The non-infected or control group of flies showed an expression of 54 proteins, while M. domestica infected with B. bassiana expressed a total of 68 hemolymph serum proteins. Thirty three proteins were expressed in both groups of houseflies, whereas 35 proteins were exclusively expressed in infected flies and 21 proteins were exclusively expressed in control flies. Among the 33 proteins which were expressed in both groups of houseflies, 17 proteins showed downregulation, while16 proteins were upregulated in the infected flies compared to the non-infected ones. The results from this study are expected to facilitate better understanding of insect's immune response mechanism.

  1. Biological trait analysis and stability of lambda-cyhalothrin resistance in the house fly, Musca domestica L. (Diptera: Muscidae).

    Science.gov (United States)

    Abbas, Naeem; Shah, Rizwan Mustafa; Shad, Sarfraz Ali; Iqbal, Naeem; Razaq, Muhammad

    2016-05-01

    House flies, Musca domestica L., (Diptera: Muscidae), are pests of poultry and have the ability to develop resistance to insecticides. To design a strategy for resistance management, life history traits based on laboratory observations were established for lambda-cyhalothrin-resistant, susceptible and reciprocal crosses of M. domestica strains. Bioassay results showed that the lambda-cyhalothrin-selected strain developed a resistance ratio of 98.34 compared to its susceptible strain. The lambda-cyhalothrin-selected strain had a relative fitness of 0.26 and lower fecundity, hatchability, lower number of next generation larvae, and net reproductive rate compared with its susceptible strain. Mean population growth rates, such as intrinsic rate of population increase, and biotic potential were lower for the lambda-cyhalothrin-selected strain compared to its susceptible strain. Resistance to lambda-cyhalothrin, indoxacarb, and abamectin was unstable while resistance to bifenthrin and methomyl was stable in the lambda-cyhalothrin-selected strain of M. domestica. Development of resistance can cost considerable fitness for the lambda-cyhalothrin-selected strain. The present study provided useful information for making potential management strategies to delay resistance development in M. domestica.

  2. The Effects of Temperature and Innate Immunity on Transmission of Campylobacter jejuni (Campylobacterales: Campylobacteraceae) Between Life Stages of Musca domestica (Diptera: Muscidae)

    DEFF Research Database (Denmark)

    Bahrndorff, Simon; Gill, C.; Lowenberger, C.

    2014-01-01

    The house fly (Musca domestica L.) is a well-established vector of human pathogens, including Campylobacter spp., which can cause infection of broiler chicken flocks, and through contaminated broiler meat can cause outbreaks of campylobacteriosis in humans. We investigated whether Campylobacter j...

  3. Chemical composition and insecticidal property of Myrsine stolonifera (Koidz.) walker (Family: Myrsinaceae) on Musca domestica (Diptera: Muscidae).

    Science.gov (United States)

    Wang, Xue Gui; Li, Qian; Jiang, Su Rong; Li, Pei; Yang, Ji Zhi

    2017-06-01

    Musca domestica is one of the most important pests of human health, and has developed strong resistance to many chemicals used for its control. One important approach for creating new pesticides is the exploration of novel compounds from plants. During a wide screening of plants with insecticidal properties that grow in southern China, we found that the methanolic extracts of Myrsine stolonifera had insecticidal activity against the adults of M. domestica. However, the insecticidal constituents and mechanisms of the M. stolonifera extracts remain unclear. The insecticidal components of the methanolic extracts of M. stolonifera were isolated with activity-guided fractionation. From the spectra of nuclear magnetic resonance (NMR) and mass spectrometry (MS), the compounds were identified as syringing (1), 2,6-dimethoxy-4-hydroxyphenol-1-O-β-d-glu (2), kaempferol-3-O-glu-rha-glu (3), and quercetin-3-O-glu-rha-glu (4). This study is the first to report the spectral data for compounds 3 and 4, and their LC 50 values were 0.52mg/g sugar and 0.36mg/g sugar 24h after treatment of the adults of M. domestica, respectively. Compounds 3 and 4 (LC 25 ) also inhibited the activities of the enzymes carboxylesterase, glutathione S-transferase, mixed function oxidase, and acetylcholine esterase of adult M. domestica, particularly mixed function oxidase and acetylcholine esterase. The cytotoxic effects of compounds 3 and 4 on cell proliferation, mitochondrial membrane potentials (MMP) and reactive oxygen species (ROS) were demonstrated on SL-1 cells. From the extracts of M. stolonifera, quercetin-3-O-glu-rha-glu and kaempferol-3-O-glu-rha-glu have displayed comparable toxicities to rotenone on M. domestica and also exhibited cytotoxic effects on SL-1 cells; therefore, the extracts of M. stolonifera and their compounds have potential as botanical insecticides to control M. domestica. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Enterobacteria associated with houseflies (Musca domestica) as an infection risk indicator in swine production farms.

    Science.gov (United States)

    Cervelin, V; Fongaro, G; Pastore, J B; Engel, F; Reimers, M A; Viancelli, A

    2018-04-23

    Houseflies (Musca domestica) spend part of their life development on animal or human manure. Manure is high in pathogenic microbes; thus, houseflies have been known as a mechanical vector for various important zoonotic diseases. Therefore, the present study showcases captured houseflies from intensive swine production regions (which are areas of high manure concentration) in Southern Brazil, and analyses their bodies' to the presence of Escherichia coli and Salmonella sp. and the sensitivity of these bacteria to various antibiotics. Additionally, Quantitative Microbiology Risk Assessment was performed simulating the contamination of lettuce by flies' bacteria and subsequent lettuce consumption by an adult human being. Houseflies were captured in swine buildings and farm houses from five farms. E. coli quantification values ranged from 10 4 to 10 6 CFU/20 flies, and all sampling sites had positive results from bacteria presence in the collected houseflies. On the other hand, Salmonella sp. presence was observed in only three farms, where the quantification ranged from 10 2 to 10 5 CFU/20 flies. The bacteria showed to be resistant to at least two from the four tested antibiotics (ampicillin, Cefalotin, Ciprofloxacin and Norfloxacin) antibiotics used in human or veterinary medicine. Infection probability analyses showed risk of human infection by E.coli, indicating possible transmission of zoonotic pathogens through flies. In this context, it was possible to conclude that there is a need for flies control, especially in swine farms where zoonotic pathogens can be abundant, to minimize the health impact of the vectorization of enteric bacteria. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. Efficacy of entomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) on developmental stages of house fly, Musca domestica.

    Science.gov (United States)

    Archana, M; D'Souza, Placid E; Patil, Jagadeesh

    2017-09-01

    The housefly, Musca domestica is a major domestic, medical and veterinary pest. The management of these flies reliance on insecticide, causes environmental constraints, insecticide resistance and residues in the meat, skin. Therefore one of the eco-friendly alternate methods is by using biological agents such as entomopathogenic nematodes (EPN). In the present study evaluated the survival of EPN species Steinernema feltiae , Heterorhabditis indica , S. carpocapsae , S. glaseri and S. abbasi in poultry manure and also their efficacy against different developmental stages of house fly. After exposing to poultry manure, S. feltiae showed more survival as followed by H. indica , S. carpocapsae , S. glaseri and S. abbasi in all exposition period. When the exposition period extended to 96 h, all nematode species survivability was drastically reduced. After exposing these nematodes to poultry manure at 24 h their virulence capacity against wax moth, Galleria mellonella showed all the nematode species were able cause 100% mortality. However their progeny production was significantly reduced. Fly eggs and pupae were refractory to these nematode infection. Petri dish without artificial diet assay showed that, second and 3rd-instar larvae were highly susceptible to EPNs as compared to larvae provided with artificial diet. H. indica showed high virulence capacity compared to other nematodes tested. Poultry manure assay revealed that, H. indica and S. carpocapsae caused minimal mortality where as S. feltiae , S. glaseri and S. abbasi did not cause any mortality. This may be because of poor survival and limited movement of nematodes in poultry manure which may be due to ammonia, other toxic substances in poultry manure. The decrease in larval mortality in manure suggests that biocontrol of housefly by using EPNs is unlikely.

  6. The blow fly, Chrysomya megacephala, and the house fly, Musca domestica, as mechanical vectors of pathogenic bacteria in Northeast Thailand.

    Science.gov (United States)

    Chaiwong, T; Srivoramas, T; Sueabsamran, P; Sukontason, K; Sanford, M R; Sukontason, K L

    2014-06-01

    The Oriental latrine fly, Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) and the house fly, Musca domestica L., (Diptera: Muscidae) are synanthropic flies which are adapted to live in close association with human habitations, thereby making them likely mechanical vectors of several pathogens to humans. There were two main aims of this study. The first aim was to determine the prevalence of these two fly species from five types of human habitations including: fresh-food markets, garbage piles, restaurants, school cafeterias and paddy fields, in the Muang Ubon Ratchathani and Warinchamrap districts of Ubon Ratchathani province of Northeast Thailand. Flies collection were conducted monthly from September 2010-October 2011 using a reconstructable funnel trap, containing 1 day-tainted beef offal as bait. A total of 7 750 flies (6 401 C. megacephala and 1 349 M.domestica) were collected. The second aim was to examine the potential of these flies to carry pathogenic bacteria. Bacteria were isolated from 994 individual flies collected using a sweep net (555 C. megacephala and 439 M. domestica). A total of 15 bacterial genera were isolated from the external surfaces, comprising ten genera of gram-negative bacteria and five gram-positive bacteria. The most common bacteria isolated from both species were coagulase-negative staphylococci, followed by Streptococcus group D non-enterococci. Human pathogenic enteric bacteria isolated were Salmonella sp., Shigella sp., Escherichia coli O157:H7, Salmonella typhi, Bacillus sp., and Enterococcus sp., of which S. typhi is the first report of isolation from these fly species. Other human pathogens included Staphylococcus aureus and Pseudomonas aeruginosa. Not only were the number of C. megacephala positive for bacteria significantly higher than for M. domestica, but they were also carrying ~11-12 times greater bacterial load than M. domestica. These data suggest that both fly species should be considered potential

  7. The Effects of Sex-Ratio and Density on Locomotor Activity in the House Fly, Musca domestica

    Science.gov (United States)

    Bahrndorff, Simon; Kjærsgaard, Anders; Pertoldi, Cino; Loeschcke, Volker; Schou, Toke M.; Skovgård, Henrik; Hald, Birthe

    2012-01-01

    Although locomotor activity is involved in almost all behavioral traits, there is a lack of knowledge on what factors affect it. This study examined the effects of sex—ratio and density on the circadian rhythm of locomotor activity of adult Musca domestica L. (Diptera: Muscidae) using an infra—red light system. Sex—ratio significantly affected locomotor activity, increasing with the percentage of males in the vials. In accordance with other studies, males were more active than females, but the circadian rhythm of the two sexes was not constant over time and changed during the light period. There was also an effect of density on locomotor activity, where males at intermediate densities showed higher activity. Further, the predictability of the locomotor activity, estimated as the degree of autocorrelation of the activity data, increased with the number of males present in the vials both with and without the presence of females. Overall, this study demonstrates that locomotor activity in M. domestica is affected by sex—ratio and density. Furthermore, the predictability of locomotor activity is affected by both sex—ratio, density, and circadian rhythm. These results add to our understanding of the behavioral interactions between houseflies and highlight the importance of these factors when designing behavioral experiments using M. domestica.

  8. Studies on the Utilization, Metabolism and Function of Sterols in the House-Fly, Musca Domestica; Utilisation. Metabolisme et fonctions des sterols chez la mouche domestique (Musca Domestica); Izuchenie usvoeniya, metabolizma i funktsii sterinov v organizme domashnej mukhi Musca Domestica; Estudios sobre la asimilacion, el metabolismo y la funcion de los esteroles in la mosca comun (Musca Domestica)

    Energy Technology Data Exchange (ETDEWEB)

    Robbins, W. E. [United States Department of Agriculture, Agricultural Research Service, Entomology Research Division, Beltsville, MD (United States)

    1963-09-15

    Insects generally have been found to require a dietary source of sterol for normal larval growth and metamorphosis. Our work has pointed to two additional physiological roles for sterols in the housefly, Musca domestica L.: (1) A dietary source of sterol is essential for sustained viable egg production in the female fly; on a sterol-deficient diet eggs are produced but hatch and viability are low. (2) Cholesterol is also involved in the mobilization and utilization of nutrient reserves associated with the initiation of ovarian maturation in the female fly. The quantitative sterol requirements for the above physiological processes and the metabolic conversions that occur during growth, metamorphosis and reproduction have been studied in this insect, using C{sup 14}- and H{sup 3}-labelled sterols in conjunction with a variety of analytical tools, including reverse isotope dilution, gasliquid chromatography and spectroscopy, and employing aseptic rearing techniques and semi-defined larval and Adult diets. Both C{sup 14}-cholesterol and H{sup 3}-{beta}- sitosterol have been used as a source df sterol in either the larval or the adult diet of the house fly, and the pattern of utilization and metabolism was found to be almost identical for these two sterols. However, there was no detectable conversion of {beta}-sitosterol to cholesterol. Sub-minimal quantities of cholesterol have also been used in the larval diet in combination with ''sparing sterols'' such as choies tanol, which will fulfill in part but not entirely the sterol requirement of this insect. The utilization and fate of the 'sparing sterol' has been investigated using C{sup 14} cholestanol, and the metabolism of the minute quantity of essential cholesterol is currently under study using high-specific-activity C{sup 14} cholesterol. - Other species of insects, including the German cockroach (Blattella germanica), have been examined in relation to the patterns of utilization and the metabolic pathways for

  9. Reduction of Escherichia coli, Salmonella Enteritidis and Campylobacter jejuni in poultry manure by rearing of Musca domestica fly larvae

    DEFF Research Database (Denmark)

    Nordentoft, Steen; Fischer, C.; Bjerrum, L.

    2017-01-01

    A major barrier for using animal waste as substrate for production of insects for feed or food is the concern for safety of the end products. In this study we investigated how rearing of fly larvae of Musca domestica in poultry manure influenced the counts of three pathogenic test strains...... of the larvae stage. This study provides data for evaluation of feed safety of fly larvae reared on animal waste. Furthermore suggests a potential use for reduction of these pathogens in manure........ Enteritidis, and C. jejuni was faster in manure with rearing of fly larvae than in manure without larvae; an 8 log10 reduction of all three test bacteria was observed within four days in manure with larvae; compared to manure without larvae where a 1 to 2 log10 was observed. We found no sign of propagation...

  10. Cytochrome P450 monooxygenase-mediated neonicotinoid resistance in the house fly Musca domestica L

    DEFF Research Database (Denmark)

    Markussen, Mette D K; Kristensen, Michael

    2010-01-01

    Neonicotinoids play an essential role in the control of house flies Musca domestica. The development of neonicotinoid resistance was found in two field populations. 766b was 130- and 140-fold resistant to imidacloprid and 17- and 28-fold resistant to thiamethoxam in males and females, respectively....... 791a was 22- and 20-fold resistant to imidacloprid and 9- and 23-fold resistant to thiamethoxam in males and females, respectively. Imidacloprid selection of 791a increased imidacloprid resistance to 75- and 150-fold in males and females, respectively, whereas selection with thiamethoxam had minimum...... of the imidacloprid-selected strain after neonicotinoid exposure. CYP6D1 expression was increased after neonicotinoid exposure in resistant males. CYP6D3 expression was induced in both sexes upon neonicotinoid exposure but significantly higher in females....

  11. Size relationships of different body parts in the three dipteran species Drosophila melanogaster, Ceratitis capitata and Musca domestica.

    Science.gov (United States)

    Siomava, Natalia; Wimmer, Ernst A; Posnien, Nico

    2016-06-01

    Body size is an integral feature of an organism that influences many aspects of life such as fecundity, life span and mating success. Size of individual organs and the entire body size represent quantitative traits with a large reaction norm, which are influenced by various environmental factors. In the model system Drosophila melanogaster, pupal size and adult traits, such as tibia and thorax length or wing size, accurately estimate the overall body size. However, it is unclear whether these traits can be used in other flies. Therefore, we studied changes in size of pupae and adult organs in response to different rearing temperatures and densities for D. melanogaster, Ceratitis capitata and Musca domestica. We confirm a clear sexual size dimorphism (SSD) for Drosophila and show that the SSD is less uniform in the other species. Moreover, the size response to changing growth conditions is sex dependent. Comparison of static and evolutionary allometries of the studied traits revealed that response to the same environmental variable is genotype specific but has similarities between species of the same order. We conclude that the value of adult traits as estimators of the absolute body size may differ among species and the use of a single trait may result in wrong assumptions. Therefore, we suggest using a body size coefficient computed from several individual measurements. Our data is of special importance for monitoring activities of natural populations of the three dipteran flies, since they are harmful species causing economical damage (Drosophila, Ceratitis) or transferring diseases (Musca).

  12. Sustainable production of housefly (Musca domestica) larvae as a protein-rich feed ingredient by utilizing cattle manure

    Science.gov (United States)

    Hussein, Mahmoud; Pillai, Viju V.; Goddard, Joshua M.; Park, Hui G.; Kothapalli, Kumar S.; Ross, Deborah A.; Ketterings, Quirine M.; Brenna, J. Thomas; Milstein, Mark B.; Marquis, Helene; Johnson, Patricia A.; Nyrop, Jan P.

    2017-01-01

    The common housefly, Musca domestica, is a considerable component of nutrient recycling in the environment. Use of housefly larvae to biodegrade manure presents an opportunity to reduce waste disposal while the rapidly assimilated insect biomass can also be used as a protein rich animal feed. In this study, we examine the biodegradation of dairy cattle manure using housefly larvae, and the nutritional value of the resulting larva meal as a feed ingredient. Our results demonstrated that dairy cattle manure presents a balanced substrate for larval growth, and the spent manure showed reductions in concentration of total nitrogen (24.9%) and phosphorus (6.2%) with an overall reduction in mass. Larva yield at an optimum density was approximately 2% of manure weight. Nutritional analysis of M. domestica larva meal showed values comparable to most high protein feed ingredients. Larva meal was 60% protein with a well-balanced amino acid profile, and 20% fat with 57% monounsaturated fatty acids, and 39% saturated fatty acids. Larva meal lacked any significant amount of omega-3 fatty acids. Evaluation of micronutrients in larva meal suggested that it is a good source of calcium and phosphorus (0.5% and 1.1% respectively). The nutritional value of larva meal closely matches that of fishmeal, making it a potentially attractive alternative for use as a protein-rich feed ingredient for livestock and aquaculture operations. PMID:28170420

  13. Reproduction and longevity of Supputius cincticeps (Het.: Pentatomidae fed with larvae of Zophobas confusa, Tenebrio molitor (Col.: Tenebrionidae or Musca domestica (Dip.: Muscidae

    Directory of Open Access Journals (Sweden)

    José Cola Zanuncio

    2005-09-01

    Full Text Available Reproduction and longevity of Supputius cincticeps (Stål (Heteroptera: Pentatomidae fed on Zophobas confusa Gebien, Tenebrio molitor L. (Coleoptera: Tenebrionidae or Musca domestica (L. (Diptera: Muscidae larvae were studied during two generations at 24.7 ± 1.1ºC, 70 ± 10% R.H. and 12 h of photophase. Body weight of newly-emerged adults, oviposition period, number of egg masses, total number of eggs and longevity of S. cincticeps were higher when fed on Z. confusa or T. molitor larvae than on M. domestica larvae. Regardless of diet, S. cincticeps showed better reproduction and longevity in the second generation in laboratory conditions.Foram avaliadas, em duas gerações, a reprodução e a longevidade de Supputius cincticeps (Stål (Heteroptera: Pentatomidae alimentado com larvas de Zophobas confusa Gebien, Tenebrio molitor L. (Coleoptera: Tenebrionidae ou Musca domestica (L. (Diptera: Muscidae a 24,7 ± 1,1ºC, 70 ± 10% de U.R. e fotofase de 12 h. O peso de adultos recém emergidos, o período de oviposição, o número de posturas, de ovos totais e a longevidade de fêmeas de S. cincticeps foram maiores com larvas de Z. confusa ou T. molitor que com M. domestica. Independentemente do tipo de presa, S. cincticeps mostrou melhor performance reprodutiva e longevidade na segunda geração.

  14. House Fly (Musca domestica L. (Diptera Muscidae Development in Different Types of Manure Desarrollo de la Mosca Doméstica (Musca domestica L. (Díptera Muscidae en Distintos Tipos de Estiércol

    Directory of Open Access Journals (Sweden)

    Patricia Larraín S

    2008-06-01

    Full Text Available Animal production units with different management systems can be found along the Huasco Valley, Region of Atacama, Chile. These constitute sources of house flies (Musca domestica L. and other vector fly species that cause damage to animals and nuisance problems in urban areas. In order to asses the importance of fly breeding sources, an experiment was carried out under laboratory conditions to evaluate different types of animal manure and composted swine manure. Time of larval development, larva mortality, pupa size, and weight and time of development to the adult stage were assessed. Results show that swine, poultry and calf manure produced a significantly higher number of adult flies, shorter life cycles and larger and heavier pupae. Cow, dog, goat and horse manure follow in effectiviveness for fly production. Composted swine manure was totally ineffective for domestic fly development.En algunas comunas del Valle del Huasco, Región de Atacama, Chile, se encuentran diversas explotaciones pecuarias con diferentes sistemas de manejo, las cuales constituyen focos generadores de mosca doméstica, Musca domestica L., y otras especies de moscas con importancia sanitaria y médica. Estos insectos provocan pérdidas económicas en dichos planteles afectando además el bienestar de poblaciones urbanas cercanas. Con el objetivo de cuantificar la importancia de dichos focos en la generación de mosca doméstica, se realizó un ensayo de laboratorio donde se evaluó como sustrato de desarrollo del insecto estiércol de diferentes especies animales y estiércol de cerdo compostado. La calidad de estos sustratos en la producción del insecto se evaluó a través de los siguientes parámetros biológicos: tiempo de desarrollo de larvas, mortalidad de larvas, tamaño y peso de pupas, y tiempo hasta la emergencia de moscas adultas. Los resultados indicaron que el estiércol de cerdo, gallina y ternero produce significativamente más moscas adultas, con un

  15. Microscopic investigation to determine the effect of Beauveria bassiana (Bals.) Vuill. and Cymbopogon citratus (DC.) Stapf. treatment on different life stages of Musca domestica (L.).

    Science.gov (United States)

    Mishra, Sapna; Kumar, Peeyush; Malik, Anushree

    2017-06-01

    Microscopic investigation was done to determine the effect of entomopathogenic fungi, Beauveria bassiana and essential oil of Cymbopogon citratus on different life stages of Musca domestica . Scanning electron microscopy investigation of fungal infected larvae showed sluggish movement, rigor, and failure of body to balance in water. Treated larvae also revealed varied level of cuticle shrinkage and extreme dehydration. Surface of B. bassiana infected pupae showed varied stage of mycelial growth, while the cadaver of adult fly was observed to have extensive fungal growth covering their entire body surface. The application of C. citratus oils on M. domestica larvae resulted in skin shrinkage, spinous cells proliferation and bleb formation, while the treated pupae showed high incidence of incomplete emergence and malformation in emerged adult flies. The current study establishes effect of C. citratus essential oil and B. bassiana infection on different life stages of M. domestica .

  16. Musca domestica Salivary Gland Hypertrophy Virus, a Globally Distributed Insect Virus That Infects and Sterilizes Female Houseflies

    DEFF Research Database (Denmark)

    Prompiboon, Pannipa; Lietze, Verena-Ulrike; Denton, John S S

    2010-01-01

    The housefly, Musca domestica, is a cosmopolitan pest of livestock and poultry and is of economic, veterinary, and public health importance. Populations of M. domestica are naturally infected with M. domestica salivary gland hypertrophy virus (MdSGHV), a nonoccluded double-stranded DNA virus...... that inhibits egg production in infected females and is characterized by salivary gland hypertrophy (SGH) symptoms. MdSGHV has been detected in housefly samples from North America, Europe, Asia, the Caribbean, and the southwestern Pacific. In this study, houseflies were collected from various locations......, and the polymorphism detected was correlated with geographic source. The virulence of the geographic MdSGHV isolates was evaluated by per os treatment of newly emerged and 24-h-old houseflies with homogenates of infected salivary glands. In all cases, 24-h-old flies displayed a resistance to oral infection...

  17. Biological changes in Musca domestica L., 1758 (Diptera:Muscidae), induced by gamma radiation (60 Co)

    International Nuclear Information System (INIS)

    Grecchi, M.A.

    1989-01-01

    This work was carried out in the Entomology Section of the Centre of Nuclear Energy in Agriculture (CENA), University of Sao Paulo, Piracicaba, Sao Paulo, Brazil. The objective of the present research work was to investigate some effects of gamma radiation on the various stages of M. domestica life cycle. (author)

  18. Insecticide mixtures could enhance the toxicity of insecticides in a resistant dairy population of Musca domestica L [corrected].

    Directory of Open Access Journals (Sweden)

    Hafiz Azhar Ali Khan

    Full Text Available House flies, Musca domestica L., are important pests of dairy operations worldwide, with the ability to adapt wide range of environmental conditions. There are a number of insecticides used for their management, but development of resistance is a serious problem. Insecticide mixtures could enhance the toxicity of insecticides in resistant insect pests, thus resulting as a potential resistance management tool. The toxicity of bifenthrin, cypermethrin, deltamethrin, chlorpyrifos, profenofos, emamectin benzoate and fipronil were assessed separately, and in mixtures against house flies. A field-collected population was significantly resistant to all the insecticides under investigation when compared with a laboratory susceptible strain. Most of the insecticide mixtures like one pyrethroid with other compounds evaluated under two conditions (1∶1-"A" and LC50: LC50-"B" significantly increased the toxicity of pyrethroids in the field population. Under both conditions, the combination indices of pyrethroids with other compounds, in most of the cases, were significantly below 1, suggesting synergism. The enzyme inhibitors, PBO and DEF, when used in combination with insecticides against the resistant population, toxicities of bifenthrin, cypermethrin, deltamethrin and emamectin were significantly increased, suggesting esterase and monooxygenase based resistance mechanism. The toxicities of bifenthrin, cypermethrin and deltamethrin in the resistant population of house flies could be enhanced by the combination with chlorpyrifos, profenofos, emamectin and fipronil. The findings of the present study might have practical significance for resistance management in house flies.

  19. Insecticide Mixtures Could Enhance the Toxicity of Insecticides in a Resistant Dairy Population of Musca domestica L

    Science.gov (United States)

    Khan, Hafiz Azhar Ali; Akram, Waseem; Shad, Sarfraz Ali; Lee, Jong-Jin

    2013-01-01

    House flies, Musca domestica L., are important pests of dairy operations worldwide, with the ability to adapt wide range of environmental conditions. There are a number of insecticides used for their management, but development of resistance is a serious problem. Insecticide mixtures could enhance the toxicity of insecticides in resistant insect pests, thus resulting as a potential resistance management tool. The toxicity of bifenthrin, cypermethrin, deltamethrin, chlorpyrifos, profenofos, emamectin benzoate and fipronil were assessed separately, and in mixtures against house flies. A field-collected population was significantly resistant to all the insecticides under investigation when compared with a laboratory susceptible strain. Most of the insecticide mixtures like one pyrethroid with other compounds evaluated under two conditions (1∶1-“A” and LC50: LC50-“B”) significantly increased the toxicity of pyrethroids in the field population. Under both conditions, the combination indices of pyrethroids with other compounds, in most of the cases, were significantly below 1, suggesting synergism. The enzyme inhibitors, PBO and DEF, when used in combination with insecticides against the resistant population, toxicities of bifenthrin, cypermethrin, deltamethrin and emamectin were significantly increased, suggesting esterase and monooxygenase based resistance mechanism. The toxicities of bifenthrin, cypermethrin and deltamethrin in the resistant population of house flies could be enhanced by the combination with chlorpyrifos, profenofos, emamectin and fipronil. The findings of the present study might have practical significance for resistance management in house flies. PMID:23613758

  20. The effect of Beauveria bassiana infection on cell mediated and humoral immune response in house fly, Musca domestica L.

    Science.gov (United States)

    Mishra, Sapna; Kumar, Peeyush; Malik, Anushree

    2015-10-01

    Entomopathogenic fungi that manifest infections by overcoming insect's immune response could be a successful control agent for the house fly, Musca domestica L. which is a major domestic, medical, and veterinary pest. In this study, the immune response of house fly to Beauveria bassiana infection was investigated to reveal fundamental aspects of house fly hemocyte biology, such as hemocyte numbers and size, which is poorly understood. The total hemocyte counts (THCs) in B. bassiana-infected house fly showed an initial increase (from 6 to 9 h), followed by subsequent decrease (9 to 12 h) with increase in time of infection. The THCs was slightly greater in infected flies than the non-infected ones. Insight into relative hemocyte counts depicted a significant increase in prohemocyte (PR) and decrease in granulocyte (GR) in infected house flies compared to non-infected ones. The relative cell area of hemocyte cells showed a noticeable increase in PR and intermediate cells (ICs), while a considerable reduction was observed for plasmatocyte (PL) and GR. The considerable variation in relative cell number and cell area in the B. bassiana-infected house flies indicated stress development during infection. The present study highlights changes occurring during B. bassiana invasion to house fly leading to establishment of infection along with facilitation in understanding of basic hemocyte biology. The results of the study is expected to help in better understanding of house fly immune response during fungal infection, so as to assist production of more efficient mycoinsecticides for house fly control using B. bassiana.

  1. Ganancia de peso del depredador Podisus distinctus (Heteroptera: Pentatomidae en combinaciones de las presas Tenebrio molitor (Coleoptera: Tenebrionidae y Musca domestica (Diptera: Muscidae

    Directory of Open Access Journals (Sweden)

    Fausto da Costa Matos Neto

    2004-03-01

    Full Text Available Entre las ninfas de los asopíneos usados para el control de gusanos desfoliadores en plantaciones de eucalipto, Podisus distinctus (Stal (Heteroptera: Pentatomidae representa un potencial agente de control biológico, sin embargo esta especie ha sido poco estudiada. El presente trabajo evaluó el efecto de las diferentes combinaciones de las presas Musca domestica L. (Diptera: Muscidae y Tenebrio molitor L. (Coleoptera: Tenebrionidae sobre el peso de ninfas de P. distinctus. El experimento se realizó en laboratorio do "Instituto de Biotecnologia Aplicada à Agropecuaria (BIOAGRO", a 25 ± 0.5ºC, 60 ± 10% de humedad relativa y 14 horas de fotoperiodo. Las ninfas de P. distinctus fueron individualizadas en cajas de Petri y alimentadas de acuerdo con los siguientes tratamientos: T1- larvas de M. domestica durante toda la fase ninfal; T2- larvas de M. domestica en el II estadio y de T. molitor en los III, IV y V estadios; T3- larvas de M. domestica en el II y III estadios y de T. molitor en los IV y V estadios; T4- larvas de M. domestica en el II, III y IV estadios y de T. molitor en el V estadio; T5- larvas de T. molitor en todos los estadios. Los mejores resultados de peso y ganancia de peso fueron encontrados cuando P. distinctus fue alimentado alternadamente con larvas de M. domestica y T. molitor. Cuando esse depredador fue solamente alimentado con larvas de M. domestica, presentó pesos menoresLitlle is known about Podisus distinctus (Stal (Heteroptera: Pentatomidae one of the Asopinae species with good possibilities for mass rearing and releasing against defoliator caterpillars in eucalyptus reforested areas in Brazil. We evaluated the impact of prey combinations on weight of nymphs and adults of P. distinctus. The prey were Musca domestica L. (Diptera: Muscidae and Tenebrio molitor L. (Coleoptera: Tenebrionidae. The experiment was developed under 25 ± 0.5ºC, 60 ± 10% R.H. and photophase of 14 hr, with nymphs of P. distinctus

  2. Reproduction and longevity of Supputius cincticeps (Het.: Pentatomidae) fed with larvae of Zophobas confusa, Tenebrio molitor (Col.: Tenebrionidae) or Musca domestica (Dip.: Muscidae)

    OpenAIRE

    Zanuncio,José Cola; Beserra,Eduardo Barbosa; Molina-Rugama,Adrián José; Zanuncio,Teresinha Vinha; Pinon,Tobias Baruc Moreira; Maffia,Vanessa Pataro

    2005-01-01

    Reproduction and longevity of Supputius cincticeps (Stål) (Heteroptera: Pentatomidae) fed on Zophobas confusa Gebien, Tenebrio molitor L. (Coleoptera: Tenebrionidae) or Musca domestica (L.) (Diptera: Muscidae) larvae were studied during two generations at 24.7 ± 1.1ºC, 70 ± 10% R.H. and 12 h of photophase. Body weight of newly-emerged adults, oviposition period, number of egg masses, total number of eggs and longevity of S. cincticeps were higher when fed on Z. confusa or T. mol...

  3. Isolation of fungi in Musca domestica Linnaeus, 1758 (Diptera: Muscidae captured at two natural breeding grounds in the municipality of Seropédica, Rio de Janeiro, Brazil

    Directory of Open Access Journals (Sweden)

    Sales Márcia de Senna Nunes

    2002-01-01

    Full Text Available The objective of this study was to isolate and identify fungal species found in natural association with adults of Musca domestica. The adult insects were collected from two natural breeding grounds: hog pens and an urban sanitary landfill. The isolated fungi were identified as: Aspergillus flavus (23.8%, A. niger var. niger (14.4%, Penicillium corylophilum (21.4%, P. fellutanum (11.9%, Cladosporium cladosporoides (4.7%, Fusarium sp. (4.7%, Alternaria alternata (11.9%, Curvularia brachyspora (2.4%, Mycelia sterilia (2.4% and the Mucorales order (2.4%.

  4. Isolation and sequence of cDNA encoding a cytochrome P-450 from an insecticide-resistant strain of the house fly, Musca domestica.

    OpenAIRE

    Feyereisen, R; Koener, J F; Farnsworth, D E; Nebert, D W

    1989-01-01

    A cDNA expression library from phenobarbital-treated house fly (Musca domestica) was screened with rabbit antisera directed against partially purified house fly cytochrome P-450. Two overlapping clones with insert lengths of 1.3 and 1.5 kilobases were isolated. The sequence of a 1629-base-pair (bp) cDNA was obtained, with an open reading frame (nucleotides 81-1610) encoding a P-450 protein of 509 residues (Mr = 58,738). The insect P-450 protein contains a hydrophobic NH2 terminus and a 22-res...

  5. Fungal dissemination by housefly (Musca domestica L.) and contamination of food commodities in rural areas of South Africa.

    Science.gov (United States)

    Phoku, J Z; Barnard, T G; Potgieter, N; Dutton, M F

    2016-01-18

    Several insects that act as vectors, including houseflies (Musca domestica L.), are often considered to be an important source of fungal contamination in human foods. Houseflies are also involved in the transmission of bacterial pathogens that may pose a serious hazard to human health. Thus, the rural population of South Africa, as typified by that in the Gauteng Province investigated in this study, is at high risk from fungal exposure disseminated by houseflies and it is therefore important to assess the role of flies in contaminating various food commodities. Eighty four samples of houseflies (captured from households and pit toilets) were studied for their potential to carry fungal spores into food commodities. The fungi occurring in samples of raw maize (15) and porridge (19) were also assessed. Fungal isolates were identified based on morphological characteristics by conventional identification methods. Fifteen genera of fungi were isolated and identified, of which Aspergillus, Fusarium, Penicillium, Cladosporium, Moniliella and Mucor were the most prevalent in all three sample types analysed. The incidence rates of fungal contamination per total fungal count isolated in houseflies, maize and porridge were recorded with mean fungal load of 2×10(8) CFU/ml, 1×10(7)CFU/g and 2×10(7)CFU/g respectively. Additionally, A. flavus, A. parasiticus, F. verticillioides, F. proliferatum, P. verrucosum, P. aurantiogriseum and M. suaveolens were the most frequent fungal isolates in houseflies with incidence rate of 34%, 11%, 27%, 21%, 22%, 17% and 32% respectively. F. verticillioides, A. flavus, A. niger and P. oslonii were the most prevalent species contaminating porridge and maize with incidence rate of 23%, 32%, 16% and 28% in maize samples, while incidence rates of 59%, 15% and 29% were recorded in porridge samples with the exception of F. verticillioides. The prevalence of these genera of fungi may pose serious health risks. Copyright © 2015 Elsevier B.V. All rights

  6. The effect of temperature on the toxicity of insecticides against Musca domestica L.: implications for the effective management of diarrhea.

    Directory of Open Access Journals (Sweden)

    Hafiz Azhar Ali Khan

    Full Text Available BACKGROUND: Diarrhea is an important cause of childhood mortality in developing countries like Pakistan because of unhygienic conditions, lack of awareness, and unwise use of preventive measures. Mechanical transmission of diarrheal pathogens by house flies, Musca domestica, is believed as the most effective route of diarrhea transmission. Although the use of insecticides as a preventive measure is common worldwide for the management of house flies, success of the measure could be compromised by the prevailing environmental temperature since it significantly affects toxicity of insecticides and thus their efficacy. Peaks of the house fly density and diarrheal cases are usually coincided and season specific, yet little is known about the season specific use of insecticides. METHODOLOGY/PRINCIPAL FINDINGS: To determine the temperature-toxicity relationship in house flies, the effect of post-bioassays temperature (range, 20-34°C on the toxicity of seven insecticides from organophosphate (chlorpyrifos, profenofos, pyrethroid (cypermethrin, deltamethrin and new chemical (emamectin benzoate, fipronil, spinosad classes was evaluated by using a feeding bioassay method. From 20-34°C, the toxicities of chlorpyrifos, profenofos, emamectin and fipronil increased 2.10, 2.93, 2.40 and 3.82 fold (i.e. positive temperature coefficient, respectively. Whereas, the toxicities of cypermethrin, deltamethrin and spinosad decreased 2.21, 2.42 and 3.16 fold (i.e. negative temperature coefficient, respectively. CONCLUSION/SIGNIFICANCE: These findings suggest that for the reduction in diarrheal cases, house flies should be controlled with insecticides according to the prevailing environmental temperature. Insecticides with a positive temperature coefficient may serve as potential candidates in controlling house flies and diarrhea epidemics in hot season and vice versa.

  7. Interactions between Entomopathogenic Fungus, Metarhizium Anisopliae and Sublethal Doses of Spinosad for Control of House Fly, Musca Domestica

    Directory of Open Access Journals (Sweden)

    M Sharififard

    2011-06-01

    Full Text Available Background: Metarhizium anisopliae strain IRAN 437C is one of the most virulent fungal isolates against house fly, Musca domestica. The objective of this study was to determine the interaction of this isolate with sublethal doses of spino­sad against housefly.Methods: In adult bioassay, conidia of entomopathogenic fungus were applied as inoculated bait at 105 and 107 spore per gram and spinosad at 0.5, 1 and 1.5 µg (A.I. per gram bait. In larval bioassay, conidia were applied as combina­tion of spore with larval bedding at 106 and 108 spore per gram and spinosad at sublethals of 0.002, 0.004 and 0.006 µg (AI per gram medium. Results: Adult mortality was 48% and 72% for fungus alone but ranged from 66–87% and 89–95% in combination treat­ments of 105 and 107 spore/g with sublethal doses of spinosad respectively. The interaction between 105 spore/g with sublethals exhibited synergistic effect, but in combination of 107 spore in spite of higher mortality, the interac­tion was additive. There was significant difference in LT50 among various treatments. LT50 values in all combination treat­ments were smaller than LT50 values in alone ones. Larval mortality was 36% and 69% for fungus alone but ranged from 58%–78% and 81%–100% in combination treatments of 106 and 108 spore/g medium with sublethals of spino­sad respectively. The interaction was synergistic in all combination treatments of larvae.Conclusion: The interaction between M. anispliae and spinosad indicated a synergetic effect that increased the house fly mortality as well as reduced the lethal time.

  8. Levels of Cadmium, Chromium and Lead in dumpsites soil, earthworm (Lybrodrilus Violaceous), Housefly (Musca Domestica) and dragon fly (Libellula luctosa)

    International Nuclear Information System (INIS)

    Adeniyi, A.A.; Okedeyi, O.O.; Idowu, A.B.

    2003-01-01

    Chemical analyses of cadmium, chromium and lead in dumpsites soil, earthworm (Lybrodrilus violaceous), housefly (Musca domestica) and in indigenous dragonfly (Libellula luctosa) were performed by atomic absorption spectrophotometry to estimate the degree of metal pollution in two Lagos dumpsites located at Iba Housing Estate (dumpsite A) and Soluos along LASU - Isheri road (dumpsite B). Soil pH and moisture content were also determined. Chromium was not detected (ND) in most of the samples except in the soil samples whose mean and standard deviation (SD) were 0.43 Plus minus 0.37 micro g/g and 0.23 plus minus 0.37 micro g/g, respectively for dumpsites A and B, and the earthworm samples harvested from dumpsite B (1.00 plus minus 1.41 micro g/g the cadmium levels were 4.00 plus minus 3.16 micro g/g and 7.50 plus minus 6.37 micro g/g for earthwarm; 2.86 plus minus 1.43 micro g/g and 4.29 plus minus 3.74 micro g/g for housefly, 0.75 plus minus 1.26 micro g/g and 1.25 plus minus 0.95 micro g/g for dragonfly, respectively for dumpsites A and B. However, the concentration of lead in the invertebrates were, 130.00 plus minus 112.58 micro g/g and 105.75 plus minus 94.44 micro g/g for earthworm; 145.71 plus minus 101.87 micro g/g and 225.71 plus minus 79.31 micro g/g for housefly; 165.00 plus minus 69.78 micro g/g and 85.00 plus minus 69.73 micro g/g for dragonfly respectively for dumpsites A and B. Cadmium and lead levels were found to be higher in the invertebrates harvested from the dumpsites than those collected from the non-dumpsites. The non-dumpsite values for cadmium were 1.24 plus minus 0,94 micro g/g, 0.45 plus minus 0.56 micro g/g and 0.38 plus minus 0.4 micro g/g for earthworm, housefly and dragonfly, respectively. Similarly, the non-dumpsite lead levels for earthworm, housefly and dragonfly were 23.12 plus minus 10.11 micro g/g, 20.75 plus minus 11.85 micro g/g and 33.62 plus minus 14.95 micro g/g, respectively.(author)

  9. Field Studies of Entomophthora (Zygomycetes: Entomophthorales)-Induced Behavioral Fever in Musca domestica (Diptera: Muscidae) in Denmark

    DEFF Research Database (Denmark)

    Kalsbeek, V.; Mullens, Bradley A.; Jespersen, Jørgen Brøchner

    2001-01-01

    House flies were collected over 3 days (three to five times per day) from specific sites on a dairy farm with a range of high to low temperatures. Flies were held individually to determine whether the distribution of fungus-infected (Entomophthora muscae and E. schizophorae) house flies differed ...... that behavioral fever occurs in the field for flies infected with both E. muscae and E. schizophorae and that flies can cure themselves of infection through the use of artificial heat sources....

  10. Factibilidad del empleo de hongos entomopatógenos en el control de Musca domestica l. en paisajes antropizados del Noroeste de Michoacán, México.

    OpenAIRE

    García Munguía, Carlos Alberto

    2016-01-01

    El objetivo de la presente investigación fue seleccionar aislamientos de los hongos M. anisopliae y B. bassiana capaces de colonizar y controlar adultos y larvas de Musca domestica e identificar las condiciones ambientales de temperatura y humedad relativa favorables para su empleo. B. bassiana y M. anisopliae fueron transmitidos sexualmente en M. domestica usando 1, 5 y 10 machos vírgenes de 3 dias de edad, expuestos a 6 x 108 conidias mL- 1 de hongos los cuales fueron confinados con 30 hemb...

  11. The effects of temperature and innate immunity on transmission of Campylobacter jejuni (Campylobacterales: Campylobacteraceae) between life stages of Musca domestica (Diptera: Muscidae).

    Science.gov (United States)

    Bahrndorff, S; Gill, C; Lowenberger, C; Skovgård, H; Hald, B

    2014-05-01

    The house fly (Musca domestica L.) is a well-established vector of human pathogens, including Campylobacter spp., which can cause infection of broiler chicken flocks, and through contaminated broiler meat can cause outbreaks of campylobacteriosis in humans. We investigated whether Campylobacter jejuni (Jones) could be transferred between life stages of M. domestica (larvae-pupae-adults) and determined bacterial counts of C. jejuni at different time points after bacterial exposure. C. jejuni was transmitted from infected larvae to pupae, but not to the adult stage. Infected larvae maintained at 25 degrees C had mean bacterial numbers of 6.5 +/- 0.2 SE log10 (colony forming units [CFU]/g) that subsequently dropped to 3.6 +/- 0.3 SE log10 (CFU/g) 8 h after infection. Pupae originating from infected larvae contained mean bacterial numbers of 5.3 +/- 0.1 SE log10 (CFU/g), and these numbers dropped to 4.8 +/- 0.1 SE log10 (CFU/g) 24 h after pupation. The decline in C. jejuni numbers during pupal development coincided with increased expression of antimicrobial peptides, including cecropin, diptericin, attacin, and defensin, in the larva-pupa transition stage and a later second peak in older pupae (4 or 48 h). Conversely, there was a reduced expression of the digestive enzyme, lysozyme, in pupae and adults compared with larvae.

  12. Protozoarios parásitos de importancia en salud pública transportados por Musca domestica Linnaeus en Lima, Perú

    Directory of Open Access Journals (Sweden)

    Martín Cárdenas

    2013-06-01

    Full Text Available El presente trabajo investiga los protozoarios parásitos de importancia para la salud pública transportados por Musca domestica. El estudio se llevó a cabo entre los meses de junio y diciembre de 1998, en los distritos de Comas y San Juan de Lurigancho de la provincia de Lima. Se estudiaron 3014 moscas colectadas en la basura acumulada en la vía pública de los pueblos jóvenes de los distritos mencionados. Las capturas se agruparon en lotes constituidos por un promedio de 30 moscas. Se estudiaron un total de 100 lotes. Las moscas fueron disecadas y procesadas con agua destilada estéril para obtener un macerado intestinal. Para la observación de los parásitos se utilizó el método directo simple y la coloración tricrómica de Gomori ácido resistente modificado. Se encontraron los siguientes protozoarios que parasitan al hombre: Blastocystis hominis (9,0%, Giardia lamblia (3,0%, Cryptosporidium sp. (2,0%, Cyclospora cayetanensis (1,0%, Iodamoeba bütschlii (17,0%, Endolimax nana (5,0% y Chilomastix mesnili (3,0%. Se reporta por primera vez para el Perú a B. hominis y Cyclospora cayetanensis en M. domestica.

  13. Desenvolvimento de Supputius cincticeps (Heteroptera, Pentatomidae alimentado com larvas de Zophobas confusa, Tenebrio molitor (Coleoptera, Tenebrionidae e Musca domestica (Diptera, Muscidae Development of Supputius cincticeps (Heteroptera, Pentatomidae fed with Zophobas confusa, Tenebrio molitor (Coleoptera, Tenebrionidae and Musca domestica (Diptera, Muscidae larvae

    Directory of Open Access Journals (Sweden)

    Eduardo B. Beserra

    1995-09-01

    Full Text Available Egg viability and nymphal development of the predatory bug Supputius cincticeps (Stål, 1860 were evaluated during two generations in the Biological Control Laboratory of the Núcleo de Biotecnologia Aplicada à Agropecuária (Bioagro/UFV in Viçosa (Minas Gerais, Brazil at 24.72±1.10ºC and photophase of 12 hours. Three treatments were represented by S. cincticeps fed with Zophobas confusa Gebien, 1906, Tenebrio molitor Linnaeus, 1758 and Musca domestica Linnaeus, 1758 larvae. Higher egg viability of this predator was found when the preys were Z. confusa and T. molitor, 74.46% and 80.91 %, than in M. domestica, 57.02%, but incubation period showed no differences between preys. Shorter nymphal development and higher nymphal viability were found with Z. confusa and T. molitor than with M. domestica. Higher weight increase was found for nymphs which originated males and females in the second generation specialy with the first two preys.

  14. Phenology of Spalangia endius Walker (Hymenoptera, Pteromalidae in pupae of Musca domestica Linnaeus (Diptera, Muscidae under laboratory conditions Fenologia de Spalangia endius Walker (Hymenoptera, Pteromalidae em pupas de Musca domestica Linnaeus (Diptera, Muscidae em condições de laboratório

    Directory of Open Access Journals (Sweden)

    Dani Furtado de Araújo

    2012-12-01

    Full Text Available This work describes the phenology of Spalangia endius Walker in pupae of Musca domestica Linnaeus under laboratory conditions. In order to understand the developmental cycle of Spalangia endius under laboratory conditions, 360 Musca domestica pupae aged from 24 to 48 hours were exposed to 15 S. endius pairs for a period of 24 hours at 26 ± 2ºC. These pupae were kept in a BOD incubator at the same temperature, with a relative humidity of Objetivando conhecer o ciclo de desenvolvimento de Spalangia endius sob condições de laboratório, 360 pupas de Musca domestica com idade de 24 a 48 horas foram expostas a 15 casais de S. endius por um período de 48 horas a 26 ± 2ºC. Estas pupas foram mantidas em BOD. Com mesma temperatura, umidade relativa <70% e com fotofase de 12 horas, onde diariamente dissecava-se 15 espécimes para avaliar o estágio e tempo de desenvolvimento do himenóptero. A fenologia permite concluir que S. endius apresenta um ciclo de desenvolvimento de 19 dias, cujo período de incubação foi de 24 horas, o desenvolvimento de larvas de S. endius ocorreu nos oito dias subsequentes nos quais uma série de alterações morfológicas foi observada. O estágio de pré-pupa deu-se no décimo dia onde cessa a movimentação e inicia a eliminação de mecônio. O estágio pupal ocorreu do décimo primeiro dia ao décimo nono, momento da emergência dos primeiros machos, já as fêmeas iniciaram a emergência aproximadamente 24 horas após. Estes resultados permitem avaliar aspectos da bionomia pormenorizada do desenvolvimento de S. endius para armazenar e programar a produção deste parasitóide, otimizando sua utilização como agente de controle biológico.

  15. Efectos de un Inhibidor de Síntesis de Quitina en el Control de Población de Musca Domestica en una Piara Comercial

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    Hoyos Duque Diego

    1997-12-01

    Full Text Available La Musca domestica asociada a las actividades pecuarias, se ha convertido en un grave problema económico y sanitario dadas las notables cantidades de desechos orgánicos generados por éstas. Se evaluó la actividad y eficacia de un insecticida inhibidor de la síntesis de quitina administrado en el alimento a cerdos en engorde. Para ello se conformaron tres grupos de animales, uno por tratamiento. A cada grupo se le suministró el insecticida mezclado en el alimento así: 25, 12,5 y 0 ppm. Los animales, durante 15 días, consumieron el alimento medicado. Durante los 7 días siguientes consumieron alimento sin medicar. En ambos periodos se evaluó el efecto del insecticida. Para ello, en los días 2, 6, 9, 12 y 15 del primer periodo, se sembraron muestras fecales con larvas de M. domestica. Igual se hizo a los 2,4, 6 y 8 del segundo periodo. Posteriormente se hicieron los recuentos de transformación de larvas a pupas y de eclosión de adultos, encontrándose un control efectivo tanto de la pupación como de la emergencia de adultos sin diferencia significativa entre las dos dosis utilizadas y significativa entre estas y la de O ppm. Se halló un incremento de la metahemoglobina proporcional a la dosis usada y no se detectaron, con la prueba de ICH, efectos teratogénicos. Se recomienda, para bajar picos poblacionales e implementar un programa integrado de control, el uso de 12,2 ppm, usada en periodos alternados de 7 días, excepto en cerdos de menos de 40 kilos y hembras gestantes.

  16. LD50 and inviably dose of gamma radiation for Musca domestica L., 1758 (diptera: muscidae) pupae aged 1, 2, 3 and 4 days

    International Nuclear Information System (INIS)

    Itepan, Natanael M.; Itepan, Sara E.D.Z.

    2013-01-01

    This experiment was carried out in Biological Control of Domestic Fly 'Eduardo Hiroshi Mizumoto' Laboratory at Entomology and Acarology Department (LEA/ESALQ/USP) and in Food Irradiation and Radioentomology Laboratory (LIARE/CENA/USP). The gamma radiation source that was used is a Co-60 irradiator model Gammabeam-650 of the Atomic Energy of Canada Ltd. whose activity in the beginning of the experiments was 9.8x10 13 Bq. (2,644 Ci). The lots of pupae of Musca domestica had been kept in acclimatized room with 25 ± 2 deg C of temperature and 70 ± 5% of relative humidity, until reaching the desired ages. Lots of pupae of M. domestica that had been used, gotten by the flotation process. They had been irradiated with the ages of 1, 2, 3 and 4 days. The used doses for 1 day pupae was 0 (control) 2.5, 5, 7.5, 10, 12.5, 15, 17.5 20, 22.5, 25, 27.5 and 30 Gy; for 2 days pupae: 0 (control), 20, 40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280 and 300 Gy; for 3 days pupae: 0 (control), 0, 40, 80, 120, 160, 200, 240, 280, 320 and 360 Gy; and for 4 days pupae: 0 (control), 40; 80; 120; 160, 200, 240, 280, 320, 360, 400 480 and 520 Gy. The dose rate was about 1,500Gy/hr. At these age intervals, the dose to prevent adult emergence was 25, 220, 360 and 520 Gy and the LD50 was 14.28, 128.04, 243.09 e 353.57 Gy, respectively. (author)

  17. LD50 and inviably dose of gamma radiation for Musca domestica L., 1758 (diptera: muscidae) pupae aged 1, 2, 3 and 4 days

    Energy Technology Data Exchange (ETDEWEB)

    Itepan, Natanael M., E-mail: nmitepan@ifsp.edu.br [Instituto Federal de Sao Paulo (IFSP), Piracicaba, SP (Brazil); Itepan, Sara E.D.Z., E-mail: sarazenitepan@ig.com.br [Universidade de Sao Paulo (FFCLRP/USP), Ribeirao Preto, SP (Brazil). Faculdade de Filosofia Ciencias e Letras; Arthur, Valter, E-mail: arthur@cena.usp.br [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil)

    2013-07-01

    This experiment was carried out in Biological Control of Domestic Fly 'Eduardo Hiroshi Mizumoto' Laboratory at Entomology and Acarology Department (LEA/ESALQ/USP) and in Food Irradiation and Radioentomology Laboratory (LIARE/CENA/USP). The gamma radiation source that was used is a Co-60 irradiator model Gammabeam-650 of the Atomic Energy of Canada Ltd. whose activity in the beginning of the experiments was 9.8x10{sup 13} Bq. (2,644 Ci). The lots of pupae of Musca domestica had been kept in acclimatized room with 25 ± 2 deg C of temperature and 70 ± 5% of relative humidity, until reaching the desired ages. Lots of pupae of M. domestica that had been used, gotten by the flotation process. They had been irradiated with the ages of 1, 2, 3 and 4 days. The used doses for 1 day pupae was 0 (control) 2.5, 5, 7.5, 10, 12.5, 15, 17.5 20, 22.5, 25, 27.5 and 30 Gy; for 2 days pupae: 0 (control), 20, 40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280 and 300 Gy; for 3 days pupae: 0 (control), 0, 40, 80, 120, 160, 200, 240, 280, 320 and 360 Gy; and for 4 days pupae: 0 (control), 40; 80; 120; 160, 200, 240, 280, 320, 360, 400 480 and 520 Gy. The dose rate was about 1,500Gy/hr. At these age intervals, the dose to prevent adult emergence was 25, 220, 360 and 520 Gy and the LD50 was 14.28, 128.04, 243.09 e 353.57 Gy, respectively. (author)

  18. Campylobacter jejuni in Musca domestica: An examination of survival and transmission potential in light of the innate immune responses of the house flies.

    Science.gov (United States)

    Gill, Carson; Bahrndorff, Simon; Lowenberger, Carl

    2017-08-01

    The house fly, Musca domestica, has been implicated as a vector of Campylobacter spp., a major cause of human disease. Little is known whether house flies serve as biological amplifying hosts or mechanical vectors for Campylobacter jejuni. We investigated the period after C. jejuni had been ingested by house flies in which viable C. jejuni colonies could be isolated from whole bodies, the vomitus and the excreta of adult M. domestica and evaluated the activation of innate immune responses of house flies to ingested C. jejuni over time. C. jejuni could be cultured from infected houseflies soon after ingestion but no countable C. jejuni colonies were observed > 24 h postingestion. We detected viable C. jejuni in house fly vomitus and excreta up to 4 h after ingestion, but no viable bacteria were detected ≥ 8 h. Suppression subtractive hybridization identified pathogen-induced gene expression in the intestinal tracts of adult house flies 4-24 h after ingesting C. jejuni. We measured the expression of immune regulatory (thor, JNK, and spheroide) and effector (cecropin, diptericin, attacin, defensing, and lysozyme) genes in C. jejuni-infected and -uninfected house flies using quantitative real time PCR. Some house fly factor, or combination of factors, eliminates C. jejuni within 24 h postingestion. Because C. jejuni is not amplified within the body of the housefly, this insect likely serves as a mechanical vector rather than as a true biological, amplifying vector for C. jejuni, and adds to our understanding of insect-pathogen interactions. © 2016 Institute of Zoology, Chinese Academy of Sciences.

  19. Novel cytochrome P450 (CYP6D1) and voltage sensitive sodium channel (Vssc) alleles of the house fly (Musca domestica) and their roles in pyrethroid resistance.

    Science.gov (United States)

    Pan, Jing; Yang, Chan; Liu, Yan; Gao, Qi; Li, Mei; Qiu, Xinghui

    2018-04-01

    The house fly Musca domestica is an important disease vector. Point mutation-mediated target-site insensitivity of the voltage sensitive sodium channel (VSSC) and increased detoxification mediated by cytochrome P450 (CYP6D1) overexpression have been characterized as two major mechanisms of pyrethroid resistance. In this study, genetic mutations in the Vssc and CYP6D1 genes and their contribution to pyrethroid resistance were investigated. Twelve lines of house flies homozygous for four genotypes were established. House flies carrying the VSSC 1014F mutation and overexpressing CYP6D1 had higher resistance to pyrethroids than those carrying 1014F alone. The presence of the 15-bp insert in the promoter region of the CYP6D1 gene did not necessarily result in a significant increase in CYP6D1 mRNA and pyrethroid resistance levels. A novel Vssc allele carrying two mutations (G1924D and G2004S) in combination with the classic 1014F and a novel CYP6D1 allele that is very similar to CYP6D1v1 were identified in Chinese house flies. This work demonstrates the effect of genetic mutations in CYP6D1 and Vssc on the susceptibility of house flies to pyrethroids, and verifies that 15-bp insert-containing CYP6D1 alleles have a single origin. These findings offer insights into the evolution of insecticide resistance and have implications for house fly control. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  20. Assessment of resistance risk to lambda-cyhalothrin and cross-resistance to four other insecticides in the house fly, Musca domestica L. (Diptera: Muscidae).

    Science.gov (United States)

    Abbas, Naeem; Shad, Sarfraz Ali

    2015-07-01

    Lambda-cyhalothrin, a sodium channel modulator insecticide, has been used frequently for the control of house flies, Musca domestica L. (Diptera: Muscidae) worldwide, including Pakistan. This experiment was performed to determine the selection and assessment of lambda-cyhalothrin resistance evolution along with four other insecticides. After 26 generations of selection, the lambda-cyhalothrin-selected population developed 445-fold resistance to lambda-cyhalothrin compared to the susceptible population. There was low cross-resistance to bifenthrin and very low cross-resistance to methomyl, imidacloprid, and fipronil in the lambda-cyhalothrin-selected population compared to the field population (G1). Realized heritability (h (2)) of resistance to lambda-cyhalothrin, bifenthrin, methomyl, imidacloprid, and fipronil was 0.07, 0.05, 0.01, 0.08, and 0.08, respectively. The projected rate of resistance development revealed that if 90 % house flies were selected, then a tenfold increase in lethal concentration 50 occurred after 17, 20, 159, 13, and 14 generations for lambda-cyhalothrin (h (2) = 0.07, slope = 2.09), bifenthrin (h (2) = 0.05, slope = 1.73), methomyl (h (2) = 0.01, slope = 2.52), imidacloprid (h (2) = 0.08, slope = 1.89), and fipronil (h (2) = 0.08, slope = 2.03), respectively. The results of our study concluded that the house fly has the potential to develop multiple insecticide resistances following continued selection pressure with lambda-cyhalothrin. This study will be helpful for assisting the development of resistance management strategies.

  1. Influences of nutrition on the biology and midgut histology of gamma irradiated Musca domestica L. (diptera: muscidae)

    International Nuclear Information System (INIS)

    Banditsing, C.

    1975-01-01

    The interaction between gamma irradiation and nutrition in the house fly, M. domestica, was investigated following irradiation of 2-hr-old adult virgin females mated with non-irradiated males. Radiation levels used were: 0 (non-irradiated), 250 rads, 500 rads, 750 rads, and 1,500 rads. Flies were fed diets consisting of varying ratio by weight of casein and sucrose at 1:1, 0.5:1, 1:0.5, and 0.1 respectively, and kept in the insectary at 25 +- 2 0 C and 70 percent R.H. Egg hatchability, adult emergence, body and ovary weights, survival of irradiated females, and histopathology were investigated at 4, 7, 11, and 15 days after irradiation. Survival of irradiated females was also determined at 45 days after irradiation. The results of these studies suggest that carbohydrate is important in house fly survival and repair of radiation damage

  2. Housefly (Musca domestica as Carrier of Enterotoxigenic Staphylococcus aureus in Broiler Farms in Iran: Is it Important for Public Health?

    Directory of Open Access Journals (Sweden)

    Mehdi Rashki Ghalehnoo

    2015-08-01

    Full Text Available Background: During the last decade, the prevalence of insect borne diseases due to contaminated food as well as the outbreaks of diseases due to enterotoxin-producing Staphylococcus aureus (ETSA strains has increased. Objectives: This study was conducted to determine the prevalence rate, enterotoxigenecity, and antimicrobial resistance of S. aureus isolated from M. domestica collected from the residential areas of nine districts broiler farms in Zabol, Iran. Materials and Methods: The flies were captured with a sterile nylon net and washed twice with distilled water. The contents were streaked onto selective media and S. aureus was identified using the conventional biochemical tests. Antimicrobial susceptibility testing was performed using CLSI guideline. Staphylococcal enterotoxins (SEs, SEA, SEB, SEC and SED, was detected by the reverse passive latex agglutination (PRLA method. Coagulase typing was performed with coagulase typing kit. Results: A total of 87 (17% S. aureus was isolated from 450 samples. The ability to synthesize staphylococcal enterotoxins (SEs was determined in 62 of 87 (71% isolates. SE type B was the most common enterotoxin found in the isolated S. aureus (45%, followed by SE type A (26%, SE type C (5%, SE type D (8%, SEA + SEC (7% and SEA + SEB (7%. Among the antibiotic tested, Penicillin was the most resistant antibiotic tested. Sixty seven percent of isolates belonged to coagulase type III, VI, VII, VIII, IV, and V. Conclusions: Our results suggest that S. aureus, including ETSA, is being carried by house flies such as M. domestica and may contribute to the spread of pathogenic isolates, with an impact on public health.

  3. Bacterial Contamination of Adult House Flies (Musca domestica) and Sensitivity of these Bacteria to Various Antibiotics, Captured from Hamadan City, Iran.

    Science.gov (United States)

    Nazari, Mansour; Mehrabi, Tahereh; Hosseini, Seyed Mostafa; Alikhani, Mohammad Yousef

    2017-04-01

    House flies ( Musca domestica ) have been known as a mechanical vector in spreading infectious diseases such as cholera, shigellosis, salmonellosis and skin infections. To investigate the bacterial contaminations of house flies and determine the resistance of these bacteria against various antibiotics. An analytical descriptive cross- sectional study was conducted. The study was performed from July to September 2015 in Hamadan University of Medical Sciences, Hamadan, Iran. A total number of 300 house flies were collected from four places, 75 flies from each place, including two educational hospitals belonging to Hamadan University of Medical Sciences, a fruit and vegetables center, and a livestock slaughter. The body surface of house flies was washed using the physiological sterile serum and the obtained solution was centrifuged at 2000 rpm for five minutes. The identification of bacteria was carried out using the phenotypic methods. The resistance of bacteria against various antibiotics was determined using the disk diffusion approach. Data were analysed by the employment of SPSS software package version 20.0. A total number of 394 bacterial strains were isolated from 275 house flies. The most prevalent type of bacteria was Bacillus spp which was detected in 31.1% of house flies. Moreover, Staphylococcus s pp. (22.9%), Escherichia coli (11.6%) were other prevalent species, whereas, Enterococcus s pp. was the least prevalent type of bacteria in the collected house flies. In terms of resistance to antibiotics, it was identified that bacteria extracted from house flies which were collected from hospital environments were more resistant to antibiotics compared with the resistance of bacteria extracted from house flies which were collected from non hospital environments. The maximum bacterial isolation was found in houseflies from hospital No.1 environment from around the accumulation of garbage. It is a well-known fact that house flies are a source of bacterial

  4. Progeny of Spalangia endius Walker, 1839 (Hymenoptera: Pteromalidae) in Musca domestica L., 1758 (Diptera: Muscidae) non-viable pupae irradiated by gamma Cobalt-60 and cold stored

    International Nuclear Information System (INIS)

    Itepan, Sara Eloiza Zen.

    1992-06-01

    This study was designed to test acceptability of cold stored irradiated and non-irradiated pupae of Musca domestica L., 1758 (Diptera: Muscidae) by the parasitoid Spalangia endius Walker, 1839 (Hymenoptera: Pteromalidae). The work was carried out in the laboratory of the Entomology Section of the Center of Nuclear Energy in Agriculture (CENA/USP) and in the laboratory of Biological Control of House Flies Eduardo Hiroshi Mizumoto of Entomology Department of the College of Agriculture Luiz de Queiroz (ESALQ/USP), Piracicaba, Sao Paulo, Brazil. The gamma radiation source used was a Cobalt-60 irradiator, Gamma bean-650, with an initial activity of 1,6265 x 10 4 Bq (6,746 Ci). The radiation LD 50 e LD 100 , using a dose ratio of 1457 Gy/h, for 12, 24, 36, 48, 60 and 72 hours old pupae and resulted in 15.35; 15.69; 11.30; 49.4; 142.78 and 175 Gy for LD 50 and 20, 22.5; 32.5; 175; 225 and 300 Gy for LD'1 00 respectively. Following exposure, the pupae were kept in climate chamber at 27 ± 3 0 C and 75 ± % of relative humidity, until all adults had emerged. In succession, 24, 48 and 72 hour-old pupae were exposed to gamma irradiation with a dose of 1440 Gy/hour, in their respective lethal doses. Following the exposure the pupae were stored at cool temperature (10 ± 2 0 C) for different periods of time (5, 15, 20 and 30 days). The same treatment was given for non-irradiated pupae. After the storage period the pupae were expose for 24 h to S. endius in a ratio of 1:10 parasitoid-host. It was not observed any statistical difference (Turkey 5%) in progeny of males and females emerged from irradiated and non-irradiated pupae. There was a preference of the parasitoid to pupae of 48 hours old followed by 24 and 72 hours old for storage period more than 5 days and shorter than 20 days. (author). 50 refs., 6 figs., 17 tabs

  5. Fluorescence technique application of X-ray in labeling with Mn, Sr and Cu, of the parasitoid and host: Muscidifuax uniraptor Kogan and Legner, 1970 (Hymenoptera: Pteromalidae) and Musca domestica L., 1758 (Diptera: Muscidae)

    International Nuclear Information System (INIS)

    Itepan, Natanael Marcio

    2003-01-01

    The objective of this work was to develop the methodology of the labeling adult of Musca domestica and Muscidifurax uniraptor using the elements Mn, Sr and Cu with the use of x-ray fluorescence. This work was carried out in the Laboratory of Biological Control of House Flies, 'Eduardo Hiroshi Mizumoto', of the 'Entomology, Phytopatology and Zoology Department of the College of Agriculture 'Luiz de Queiroz' ESALQ/USP), and the Division of Methods the Development and Nuclear Analytics Techniques, of CENA/USP, Piracicaba, Sao Paulo, Brazil. The larvae was removed to the labeled diet with increasing level of the elements Mn, Sr and Cu. The levels tested for all element were: 0 (control); 0,25; 0,50; 1,00; 2,00; 4,00; 8,00; 16,00; 32,00 and 64,00 mg/gr of diet. Labeled pupae with 1,00 to 4,00 (Mn) and 1,00 (Sr and Cu) mg/gr of diet were tested for the pupal parasitoid M. uniraptor. The concentration quantity of the chemical elements was realized by the Analytical Technique denominated (EDXRF) energy dispersive X-ray fluorescence. Concentrations of 2,00 (Mn) and 1,00 (Sr) supplemented to the diet of M. domestica were sufficient for the adult insect labeled, however, not affecting its life expectancy. Pupae originated from the larvae of M. domestica treated with dose of 2,00 (Mn) and 1,00 (Sr and Cu) mg supplemented to the diet, and used as hosts of the parasitoid M. uniraptor, affected the viability of the immature phase and did not label the adults. (author)

  6. Synergism between caffeine and γ-radiation in the induction of dominant lethal mutations in oocytes and spermatozoa of Musca domestica

    International Nuclear Information System (INIS)

    Targa, H.J.

    1983-01-01

    Caffeine was studied with regard to its synergism with γ-radiation in the induction of dominant lethal mutations in S14 oocytes and mature spermatozoa of M. domestica. In S14 oocytes an increase in the frequency of such a type of mutation was observed only when the exposure to γ-radiation followed a pretreatment with a diet containing 0.2% of caffeine. Negative results were obtained with (a) post-treatment with the same kind of diet, (b) pretreatment with diets containing 0.1 and 0.02% of caffeine and (c) exposure to the radiation 6 h after interruption of the feeding treatment with the diet containing 0.2% of caffeine. Such influence of the conditions under which the treatment is performed and the synergistic effects is probably related to the food intake pattern and the rapid metabolism of the caffeine. When the 0.2% caffeine pretreatment was combined with an exposure of the oocytes to variable doses of γ-radiation, the increments in the mutations observed seemed to be negatively correlated to the radiation doses used. Also, under such conditions, the dose/survival relationship fits well an exponential curve expressed by in y=-0.866chi. With mature spermatozoa, synergism by caffeine was found only when the females, after having been mated with the irradiated males, were fed for 24 h on a diet supplemented with 0.2% of caffeine. (orig.)

  7. Tolerance of house fly, Musca domestica L. (Diptera: Muscidae) to dichlorvos (76% EC) an insecticide used for fly control in the tsunami-hit coastal villages of southern India.

    Science.gov (United States)

    Srinivasan, R; Jambulingam, P; Gunasekaran, K; Boopathidoss, P S

    2008-02-01

    The Directorate of Public Health (DPH), Tamil Nadu, in southern India employed spraying of dichlorvos (76% EC) for quick elimination of fly concentrations in the tsunami-hit coastal villages at the concentration of 304g (a.i.)/10,000m(2). However, nuisance of house flies remained high particularly in temporary shelters and centralized relief kitchens. Susceptibility of house fly, Musca domestica to dichlorvos was determined in the laboratory to provide information for an effective management of this pest. Various concentrations of dichlorvos (76% EC) viz., 0.1, 0.2, 0.4, 0.6 and 0.8microg (a.i.) per fly, were tested using topical application against F(1) progenies of house flies collected 12 months after insecticide applications from different habitats in the tsunami-hit coastal villages. Fly mortality was recorded at 24h post treatment. Parallel controls were maintained for comparison. Mortality of the house flies varied between 17.5% and 100% and increased with an increase in dosage of the insecticide. Mortality was >80% at 0.6 and 0.8microg (a.i.) per fly. The LD(50) of dichlorvos tested against flies collected from different villages varied from 0.218microg (a.i.) to 0.235microg (a.i.) per fly and the LD(90) varied from 0.574microg (a.i.) to 0.639microg (a.i.) per fly. House flies collected from a rural village, Thirukanur that had never been exposed for insecticide treatment in the past one decade, when tested, the mortality varied between 92.5% and 100% and increased with concentration of dichlorvos. Mortality was >90% from 0.2microg (a.i.) per fly and the LD(50) was 0.0399microg (a.i.)/fly, while the LD(90) was 0.1604microg (a.i.)/fly. The LD(90) values of the flies collected from the tsunami-hit villages were 3.5-3.9 times higher than that of the flies collected from Thirukanur. Fly abundance remained high in tsunami-hit villages with no marked reduction, suggesting that the flies had developed tolerance to dichlorvos. It is suggested that for an effective

  8. Activity and relative abundance of hymenopterous parasitoids that attack puparia of Musca domestica and Stomoxys calcitrans (Diptera: Muscidae) on confined pig and cattle farms in Denmark

    DEFF Research Database (Denmark)

    Skovgård, H; Jespersen, J B

    1999-01-01

    (Linnaeus). In total, seven pteromalid species were recovered: Spalangia cameroni Perkins, S. nigripes Curtis, S. subpunctata Förster, Muscidifurax raptorGirault & Sanders, Pachycrepoideus vindemiae (Rondani), Urolepis rufipes(Ashmead) and Nasonia vitripennis (Walker), an ichneumonid Phygadeuon fumator......Gravenhorst, a diapriid Trichopria sp., and a staphylinid Aleocharasp. This is the first time that U. rufipes has been recorded in Europe. Spalangia cameroni and M. raptor were the most frequently recorded species in all regions of the country, and accounted for the main parasitism of Musca domesticaand Stomoxys...... calcitrans puparia. The overall rate of parasitism per farm was low: 12.9% of the total number of fly puparia collected. Direct ordination, used to assess the habitat distribution of the parasitoids, showed that Muscidifurax raptor mainly seeks fly puparia in outdoor manure heaps and especially in manure...

  9. Exuberant oral myiasis caused by Musca domestica (Housefly

    Directory of Open Access Journals (Sweden)

    Rajkumar N Parwani

    2014-01-01

    Full Text Available Tissues of oral cavity, when invaded by the parasitic larvae of houseflies, the condition is called as oral myiasis. It is a rare disease that is most common in developing countries and is associated with conditions leading to persistent mouth opening along with poor oral hygiene, suppurative lesions, severe halitosis and maxillofacial trauma. A case of exuberant oral myiasis in a 42-year-old female patient is described here. She reported with swelling, pain, mobility of teeth and foul odor. Diagnosis was based primarily on history and clinical features. Management included use of turpentine oil, mechanical removal of larvae followed by extraction of mobile teeth and curettage along with supportive antibiotic and analgesic therapy. Supportive nutritional supplements and timely institution of treatment encompassing removal of the offending larvae and carious teeth with proper education and motivation of the patient including oral hygiene instructions led to the resolution of these lesions.

  10. House Fly (Musca domestica L. Attraction to Insect Honeydew.

    Directory of Open Access Journals (Sweden)

    Kim Y Hung

    Full Text Available House flies are of major concern as vectors of food-borne pathogens to food crops. House flies are common pests on cattle feedlots and dairies, where they develop in and feed on animal waste. By contacting animal waste, house flies can acquire human pathogenic bacteria such as Escherichia coli and Salmonella spp., in addition to other bacteria, viruses, or parasites that may infect humans and animals. The subsequent dispersal of house flies from animal facilities to nearby agricultural fields containing food crops may lead to pre-harvest food contamination with these pathogens. We hypothesized that odors from honeydew, the sugary excreta produced by sucking insects feeding on crops, or molds and fungi growing on honeydew, may attract house flies, thereby increasing the risk of food crop contamination. House fly attraction to honeydew-contaminated plant material was evaluated using a laboratory bioassay. House flies were attracted to the following plant-pest-honeydew combinations: citrus mealybug on squash fruit, pea aphid on faba bean plants, whitefly on navel orange and grapefruit leaves, and combined citrus mealybug and cottony cushion scale on mandarin orange leaves. House flies were not attracted to field-collected samples of lerp psyllids on eucalyptus plants or aphids on crepe myrtle leaves. Fungi associated with field-collected honeydews were isolated and identified for further study as possible emitters of volatiles attractive to house flies. Two fungal species, Aureobasidium pullulans and Cladosporium cladosporioides, were repeatedly isolated from field-collected honeydew samples. Both fungal species were grown in potato dextrose enrichment broth and house fly attraction to volatiles from these fungal cultures was evaluated. House flies were attracted to odors from A. pullulans cultures but not to those of C. cladosporioides. Identification of specific honeydew odors that are attractive to house flies could be valuable for the development of improved house fly baits for management of this pest species.

  11. Insecticidal activity of Jatropha curcas extracts against housefly, Musca domestica.

    Science.gov (United States)

    Chauhan, Nitin; Kumar, Peeyush; Mishra, Sapna; Verma, Sharad; Malik, Anushree; Sharma, Satyawati

    2015-10-01

    The hexane and ether extracts of leaves, bark and roots of Jatropha curcas were screened for their toxicity against different developmental stages of housefly. The larvicidal, pupicidal and adulticidal activities were analysed at various concentrations (0.78-7.86 mg/cm(2)) of hexane and ether extracts. The lethal concentration values (LC50) of hexane extract of J. curcas leaves were 3.0 and 0.27 mg/cm(2) for adult and larval stages of housefly, respectively, after 48 h. Similarly, the ether extract of leaf showed the LC50 of 2.20 and 4.53 mg/cm(2) for adult and larval stages of housefly. Least toxicity was observed with hexane root extract of J. curcas with LC50 values of 14.18 and 14.26 mg/cm(2) for adult and larvae of housefly, respectively, after 48 h. The variation in LC50 against housefly pupae was found to be 8.88-13.10 mg/cm(2) at various J. curcas extract concentrations. The GC-MS analysis of J. curcas leaf extract revealed the presence of trans-phytol (60.81 %), squalene (28.58 %), phytol (2.52 %) and nonadecanone (1.06 %) as major components that could be attributed for insecticidal activity of J. curcas extracts.

  12. Dispersal, behavioral responses and thermal adaptation in Musca domestica

    DEFF Research Database (Denmark)

    Kjaersgaard, Anders; Blackenhorn, Wolf U.; Pertoldi, Cino

    were obtained with flies held for several generations in a laboratory common garden setting, therefore we suggest that exposure to and avoidance of high temperatures under natural conditions has been an important selective agent causing the suggested adaptive differentiation between the populations.......Behavioral traits can have great impact on an organism’s ability to cope with or avoidance of thermal stress, and are therefore of evolutionary importance for thermal adaptation. We compared the morphology, heat resistance, locomotor (walking and flying) activity and flight performance of three...

  13. Mosquito-larvicidal efficacy of the extract of Musca domestica ...

    African Journals Online (AJOL)

    The intolerably high burdens of mosquito-borne diseases will be reduced sustainably through the development of integral eco-friendly alternative insecticides of natural products origin. The need to broaden the global search for such insecticidal lead-agents, especially , those that will be less vulnerable to resistance, was ...

  14. Circadian rhythm gene regulation in the housefly, Musca domestica

    Czech Academy of Sciences Publication Activity Database

    Codd, V.; Doležel, David; Stehlík, Jan; Piccin, A.; Garner, K. J.; Racey, S. N.; Straatman, K. R.; Louis, E. J.; Costa, R.; Šauman, Ivo; Kyriacou, C. P.; Rosato, E.

    2007-01-01

    Roč. 177, č. 3 (2007), s. 1539-1551 ISSN 0016-6731 R&D Projects: GA ČR GA204/04/0862; GA MŠk 2B06129 Institutional research plan: CEZ:AV0Z50070508 Keywords : circadian * evolution * Diptera Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.001, year: 2007

  15. House Fly (Musca domestica L.) Attraction to Insect Honeydew

    Science.gov (United States)

    Hung, Kim Y.; Michailides, Themis J.; Millar, Jocelyn G.; Wayadande, Astri; Gerry, Alec C.

    2015-01-01

    House flies are of major concern as vectors of food-borne pathogens to food crops. House flies are common pests on cattle feedlots and dairies, where they develop in and feed on animal waste. By contacting animal waste, house flies can acquire human pathogenic bacteria such as Escherichia coli and Salmonella spp., in addition to other bacteria, viruses, or parasites that may infect humans and animals. The subsequent dispersal of house flies from animal facilities to nearby agricultural fields containing food crops may lead to pre-harvest food contamination with these pathogens. We hypothesized that odors from honeydew, the sugary excreta produced by sucking insects feeding on crops, or molds and fungi growing on honeydew, may attract house flies, thereby increasing the risk of food crop contamination. House fly attraction to honeydew-contaminated plant material was evaluated using a laboratory bioassay. House flies were attracted to the following plant-pest-honeydew combinations: citrus mealybug on squash fruit, pea aphid on faba bean plants, whitefly on navel orange and grapefruit leaves, and combined citrus mealybug and cottony cushion scale on mandarin orange leaves. House flies were not attracted to field-collected samples of lerp psyllids on eucalyptus plants or aphids on crepe myrtle leaves. Fungi associated with field-collected honeydews were isolated and identified for further study as possible emitters of volatiles attractive to house flies. Two fungal species, Aureobasidium pullulans and Cladosporium cladosporioides, were repeatedly isolated from field-collected honeydew samples. Both fungal species were grown in potato dextrose enrichment broth and house fly attraction to volatiles from these fungal cultures was evaluated. House flies were attracted to odors from A. pullulans cultures but not to those of C. cladosporioides. Identification of specific honeydew odors that are attractive to house flies could be valuable for the development of improved house fly baits for management of this pest species. PMID:25970333

  16. House Fly (Musca domestica L.) Attraction to Insect Honeydew.

    Science.gov (United States)

    Hung, Kim Y; Michailides, Themis J; Millar, Jocelyn G; Wayadande, Astri; Gerry, Alec C

    2015-01-01

    House flies are of major concern as vectors of food-borne pathogens to food crops. House flies are common pests on cattle feedlots and dairies, where they develop in and feed on animal waste. By contacting animal waste, house flies can acquire human pathogenic bacteria such as Escherichia coli and Salmonella spp., in addition to other bacteria, viruses, or parasites that may infect humans and animals. The subsequent dispersal of house flies from animal facilities to nearby agricultural fields containing food crops may lead to pre-harvest food contamination with these pathogens. We hypothesized that odors from honeydew, the sugary excreta produced by sucking insects feeding on crops, or molds and fungi growing on honeydew, may attract house flies, thereby increasing the risk of food crop contamination. House fly attraction to honeydew-contaminated plant material was evaluated using a laboratory bioassay. House flies were attracted to the following plant-pest-honeydew combinations: citrus mealybug on squash fruit, pea aphid on faba bean plants, whitefly on navel orange and grapefruit leaves, and combined citrus mealybug and cottony cushion scale on mandarin orange leaves. House flies were not attracted to field-collected samples of lerp psyllids on eucalyptus plants or aphids on crepe myrtle leaves. Fungi associated with field-collected honeydews were isolated and identified for further study as possible emitters of volatiles attractive to house flies. Two fungal species, Aureobasidium pullulans and Cladosporium cladosporioides, were repeatedly isolated from field-collected honeydew samples. Both fungal species were grown in potato dextrose enrichment broth and house fly attraction to volatiles from these fungal cultures was evaluated. House flies were attracted to odors from A. pullulans cultures but not to those of C. cladosporioides. Identification of specific honeydew odors that are attractive to house flies could be valuable for the development of improved house fly baits for management of this pest species.

  17. Oggetti e memoria domestica

    Directory of Open Access Journals (Sweden)

    Marina Brancato

    2011-12-01

    Full Text Available Questo contributo è un ideale viaggio nella memoria attraverso le stanze che compongono le nostre case. Un itinerario fra gli oggetti che fanno da sfondo quotidiano alla nostra scenografia domestica. Nello specifico indagheremo album fotografici, souvenir, ninnoli (mnemoteche domestiche. Sin dall’inizio abbiamo considerato gli ambienti domestici veri e propri serbatoi di memorie individuali e collettive. In tali serbatoi i soggetti depositano tracce delle proprie biografie, esprimendo e confermando progetti identitari.

  18. Distribution and substrate specificity of esterases in the housefly, Musca domestica L.

    NARCIS (Netherlands)

    Asperen, K. van

    1959-01-01

    Housefly homogenates perform high cholinesterase and ali-esterase activity. Warburg-manometric studies show that acetylcholine, acetyl-β-methylcholine, butyrylcholine, and benzoylcholine are exclusively hydrolysed by a cholinesterase, the properties of which are more or less comparable to those of

  19. Effect of temperature and humidity on pathogenicity of native Beauveria bassiana isolate against Musca domestica L.

    Science.gov (United States)

    Mishra, Sapna; Kumar, Peeyush; Malik, Anushree

    2015-12-01

    Beauveria bassiana HQ917687 virulence to housefly larvae and adult was assessed at different relative humidity, RH (50, 75, 90, and 100 %) and temperature (15, 20, 25, 30, 35, 40, 45 °C) conditions at the fungal dose of 10(8) conidia/ml. Depending on the temperature and RH regime tested, difference in mortality rates of housefly adult and larvae were detected. During assay on adult housefly, 100 % mortality was achieved at RH, 90 and 100 % while the temperature of 30 °C showed maximum mortality at all the tested humidity conditions. Lethal time, LT50 was 2.9 days at 100 % RH. Larval mortality at different humidity conditions varied between 30 and 74 %, with maximum mortality at 100 % RH and 30 °C. Optimum temperature for B. bassiana virulence to housefly larvae was also found to be 30 °C. The interaction between temperature and RH revealed significant effect of RH at moderate temperature range (20-35 °C), while such an interaction was not observed at extreme temperatures. The results obtained in this study have useful implications in understanding the pathogen behavior under actual field conditions. This in turn may help devising suitable entomopathogen release schedules for maximum fungal infection.

  20. Composting poultry manure by fly larvae (Musca domestica) eliminates Campylobacter jejuni from the manure

    DEFF Research Database (Denmark)

    Nordentoft, Steen; Hald, Birthe

    2013-01-01

    study To monitor fly larvae composting of poultry manure artificially contaminated with C. jejuni, and to investigate a possible transmission route of C. jejuni from the manure through the fly larvae to the adult fly. Conclusions The addition of fly larvae both accelerated the degradation of manure...

  1. The gut microbiota in larvae of the housefly Musca domestica and their horizontal transfer through feeding.

    Science.gov (United States)

    Zhao, Yao; Wang, Wanqiang; Zhu, Fen; Wang, Xiaoyun; Wang, Xiaoping; Lei, Chaoliang

    2017-12-01

    House fly larvae provide a prolific and sustainable source of proteins used in poultry and fish feed. Wheat bran is a superior diet for house fly larvae and has been widely investigated to exploit its potential in the food and feed area. Using Illumina MiSeq 16S rDNA sequencing, this study investigated the gut microbiota of house fly larvae feeding on wheat bran and the bacterial community in the wheat bran. The bacterial communities in the house fly larvae were dominated by the phyla Proteobacteria and Firmicutes. Enterobacteriaceae and Providencia were the predominant bacteria at the family and genus levels, respectively. Some bacteria in the phyla Actinobacteria, Proteobacteria, Bacteroidetes and Firmicutes may be transferred from the gut of house flies to the wheat bran during feeding and may be involved in degrading and utilizing polysaccharides in the cell wall of wheat bran. The significance of the gut microbiota of house fly larvae, their transferring and roles in degradation of wheat bran is discussed. These findings regarding the gut microbiota of house fly larvae will provide opportunities for research on the impact of microbial communities on poultry and fish.

  2. Susceptibility to thiamethoxam of Musca domestica from Danish livestock farms

    DEFF Research Database (Denmark)

    Kristensen, Michael; Jespersen, Jørgen B

    2008-01-01

    . There was no correlation between the toxicities of thiamethoxam and spinosad, dimethoate, methomyl, bioresmethrin or azamethiphos. The toxicity in feeding bioassay at 72 h of imidacloprid in a susceptible strain was 32 μg imidacloprid g-1 sugar at LC50, making it 19-fold less toxic to houseflies than thiamethoxam....... There was a strong significant correlation between the toxicities of thiamethoxam and imidacloprid in field populations. CONCLUSION: Neonicotinoid-resistant houseflies were present at a detectable and noticeable level before thiamethoxam and imidacloprid were introduced for housefly control in Denmark. The toxicity...... of thiamethoxam is explained by other parameters than the toxicities of spinosad, dimethoate, methomyl, bioresmethrin or azamethiphos. The cross-resistance between thiamethoxam and imidacloprid indicates a coincidence of mechanism of the toxicity and resistance in the field populations....

  3. A comparative study of P450 gene expression in field and laboratory Musca domestica L. strains.

    Science.gov (United States)

    Højland, Dorte H; Vagn Jensen, Karl-Martin; Kristensen, Michael

    2014-08-01

    The housefly is a global pest that has developed resistance to most insecticides applied for its control. Resistance has been associated with cytochrome P450 monooxygenases (P450s). The authors compare the expression of six genes possibly associated with insecticide resistance in three unselected strains: a multiresistant strain (791a), a neonicotinoid-resistant strain (766b) and a new field strain (845b). CYP4G2 was highly expressed throughout the range of strains and proved to be the one of the most interesting expression profiles of all P450s analysed. CYP6G4 was expressed up to 11-fold higher in 766b than in WHO-SRS. Significant differences between expression of P450 genes between F1 flies from 845b and established laboratory strains were shown. In general, P450 gene expression in 845b was 2-14-fold higher than in the reference strain (P resistance. There is a strong indication that CYP6G4 is a major insecticide resistance gene involved in neonicotinoid resistance. © 2013 Society of Chemical Industry.

  4. A comparative study of P450 gene expression in field and laboratory Musca domestica L. strains

    DEFF Research Database (Denmark)

    Højland, Dorte Heidi; Vagn Jensen, Karl-Martin; Kristensen, Michael

    2014-01-01

    BACKGROUND The housefly is a global pest that has developed resistance to most insecticides applied for its control. Resistance has been associated with cytochrome P450 monooxygenases (P450s). The authors compare the expression of six genes possibly associated with insecticide resistance in three...... unselected strains: a multiresistant strain (791a), a neonicotinoid-resistant strain (766b) and a new field strain (845b). RESULTS CYP4G2 was highly expressed throughout the range of strains and proved to be the one of the most interesting expression profiles of all P450s analysed. CYP6G4 was expressed up...... to 11-fold higher in 766b than in WHO-SRS. Significant differences between expression of P450 genes between F1 flies from 845b and established laboratory strains were shown. In general, P450 gene expression in 845b was 2–14-fold higher than in the reference strain (P

  5. A Case for Sequencing the Genome of Musca domestica (Diptera: Muscidae)

    DEFF Research Database (Denmark)

    Scott, J G; Liu, N; Kristensen, M

    2009-01-01

    House flies are carriers of >100 devastating diseases that have severe consequences for human and animal health. Despite the fact that it is a passive vector, a key bottleneck to progress in controlling the human diseases transmitted by house flies is lack of knowledge of the basic molecular...

  6. Acetylcholinesterase Inhibitory Activity of Green Tea Polyphenols ...

    African Journals Online (AJOL)

    Inhibition of acetylcholinesterase activity is one of the most popular approaches for treatment of neurological disorders such as Alzheimer's disease and others. In the present study, we evaluated inhibition of acetylcholinesterase activity by different concentrations of green tea (Camellia sinensis L.) extract using ...

  7. Filamentary structure and magnetic field orientation in Musca

    Science.gov (United States)

    Cox, N. L. J.; Arzoumanian, D.; André, Ph.; Rygl, K. L. J.; Prusti, T.; Men'shchikov, A.; Royer, P.; Kóspál, Á.; Palmeirim, P.; Ribas, A.; Könyves, V.; Bernard, J.-Ph.; Schneider, N.; Bontemps, S.; Merin, B.; Vavrek, R.; Alves de Oliveira, C.; Didelon, P.; Pilbratt, G. L.; Waelkens, C.

    2016-05-01

    Herschel has shown that filamentary structures are ubiquitous in star-forming regions, in particular in nearby molecular clouds associated with Gould's Belt. High dynamic range far-infrared imaging of the Musca cloud with SPIRE and PACS reveals at least two types of filamentary structures: (1) the main ~10-pc scale high column-density linear filament; and (2) low column-density striations in close proximity to the main filament. In addition, we find features with intermediate column densities (hair-like strands) that appear physically connected to the main filament. We present an analysis of this filamentary network traced by Herschel and explore its connection with the local magnetic field. We find that both the faint dust emission striations and the plane-of-the-sky (POS) magnetic field are locally oriented close to perpendicular to the high-density main filament (position angle ~25-35°). The low-density striations and strands are oriented parallel to the POS magnetic field lines, which are derived previously from optical polarization measurements of background stars and more recently from Planck observations of dust polarized emission. The position angles are 97 ± 25°, 105 ± 7°, and 105 ± 5°. From these observations, we propose a scenario in which local interstellar material in this cloud has condensed into a gravitationally-unstable filament (with "supercritical" mass per unit length) that is accreting background matter along field lines through the striations. We also compare the filamentary structure in Musca with what is seen in similar Herschel observations of the Taurus B211/3 filament system and find that there is significantly less substructure in the Musca main filament than in the B211/3 filament. We suggest that the Musca cloud may represent an earlier evolutionary stage in which the main filament has not yet accreted sufficient mass and energy to develop a multiple system of intertwined filamentary components. Herschel is an ESA space

  8. A novel endogenous inhibitor of phenoloxidase from Musca domestica has a cystine motif commonly found in snail and spider toxins.

    Science.gov (United States)

    Daquinag, A C; Sato, T; Koda, H; Takao, T; Fukuda, M; Shimonishi, Y; Tsukamoto, T

    1999-02-16

    Phenoloxidase inhibitor (POI), found in the hemolymph of housefly pupae, is a novel dopa-containing and cystine-rich peptide that competitively inhibits phenoloxidase with a Ki in the nanomolar range. [Tyr32]POI is a potential precursor molecule also found in the hemolymph that may be posttranslationally oxidized to the dopa-containing peptide after creation of a rigid structure. By employing both a solid-phase peptide synthesis system based on a 9-fluorenylmethoxycarbonyl strategy and a specific air oxidation technique to ensure correct folding, we have been able to synthesize [Tyr32]POI. The synthetic [Tyr32]POI was confirmed to be identical to the native [Tyr32]POI by coelution high-performance liquid chromatography analysis and by enzymatic analysis using the phenoloxidase inhibition assay. To determine the disulfide pairings within the peptides, a series of enzyme hydrolyses and partial reduction/alkylation steps were performed. Three cystine pairs (Cys11-Cys25, Cys18-Cys29, and Cys24-Cys36) were determined by identification of the resulting peptides. The disulfide pairings of the two adjacent Cys residues (Cys11-Cys25 and Cys24-Cys36) were unambiguously assigned by comparing the derived fragments with the two possible isomers synthesized through a novel disulfide-linking technique. The arrangement of the disulfide bridges in POI was found to be topologically identical to those found for several peptides within the inhibitor cystine knot structural family. Although these peptides share a low primary sequence homology and display a diversity of biological functions, they nonetheless share similarities in their cystine motifs and tertiary structure. The tertiary structure model of POI, which was derived through molecular dynamics and energy minimization studies using restraints with determined disulfide connectivities, suggests that POI is a new class member of the inhibitor cystine-knot structural family.

  9. Bioconversion of Gibberellin Fermentation Residue into Feed Supplement and Organic Fertilizer Employing Housefly (Musca domestica L. Assisted by Corynebacterium variabile.

    Directory of Open Access Journals (Sweden)

    Sen Yang

    Full Text Available The accumulation of a considerable quantity of gibberellin fermentation residue (GFR during gibberellic acid A3 (GA3 production not only results in the waste of many resources, but also poses a potential hazard to the environment, indicating that the safe treatment of GFR has become an urgent issue for GA3 industry. The key to recycle GFR is converting it into an available resource and removing the GA3 residue. To this end, we established a co-bioconversion process in this study using house fly larvae (HFL and microbes (Corynebacterium variabile to convert GFR into insect biomass and organic fertilizer. About 85.5% GA3 in the GFR was removed under the following optimized solid-state fermentation conditions: 60% GFR, 40% rice straw powder, pH 8.5 and 6 days at 26 °C. A total of 371 g housefly larvae meal and 2,064 g digested residue were bio-converted from 3,500 g raw GFR mixture contaning1, 400 g rice straw in the unit of (calculated dry matter. HFL meal derived from GFR contained 56.4% protein, 21.6% fat, and several essential amino acids, suggesting that it is a potential alternative animal feed protein source. Additionally, the digested GFR could be utilized as an organic fertilizer with a content of 3.2% total nitrogen, 2.0% inorganic phosphorus, 1.3% potassium and 91.5% organic matter. This novel GFR bio-conversion method can mitigate potential environmental pollution and recycle the waste resources.

  10. Bioconversion of Gibberellin Fermentation Residue into Feed Supplement and Organic Fertilizer Employing Housefly (Musca domestica L.) Assisted by Corynebacterium variabile

    Science.gov (United States)

    Yang, Sen; Xie, Jiufeng; Hu, Nan; Liu, Yixiong; Zhang, Jiner; Ye, Xiaobin; Liu, Ziduo

    2015-01-01

    The accumulation of a considerable quantity of gibberellin fermentation residue (GFR) during gibberellic acid A3 (GA3) production not only results in the waste of many resources, but also poses a potential hazard to the environment, indicating that the safe treatment of GFR has become an urgent issue for GA3 industry. The key to recycle GFR is converting it into an available resource and removing the GA3 residue. To this end, we established a co-bioconversion process in this study using house fly larvae (HFL) and microbes (Corynebacterium variabile) to convert GFR into insect biomass and organic fertilizer. About 85.5% GA3 in the GFR was removed under the following optimized solid-state fermentation conditions: 60% GFR, 40% rice straw powder, pH 8.5 and 6 days at 26°C. A total of 371g housefly larvae meal and 2,064g digested residue were bio-converted from 3,500g raw GFR mixture contaning1, 400g rice straw in the unit of (calculated) dry matter. HFL meal derived from GFR contained 56.4% protein, 21.6% fat, and several essential amino acids, suggesting that it is a potential alternative animal feed protein source. Additionally, the digested GFR could be utilized as an organic fertilizer with a content of 3.2% total nitrogen, 2.0% inorganic phosphorus, 1.3% potassium and 91.5% organic matter. This novel GFR bio-conversion method can mitigate potential environmental pollution and recycle the waste resources. PMID:25992605

  11. Continuous recording of excretory water loss from Musca domestica using a flow-through humidity meter: hormonal control of diuresis.

    Science.gov (United States)

    Coast, Geoffrey M

    2004-05-01

    Water loss from adult male houseflies was continuously recorded using a flow-through humidity meter, which enabled losses to be apportioned between the sum of cuticular and respiratory transpiration, salivation and excretion. Transpiration accounted for >95% of water lost from sham-injected flies, compared with excretion (3.0%) and salivation (2.4%). In contrast, excretion accounted for 40% of water lost from flies injected with > or =3 microl of saline, whereas salivary losses were unchanged. Saline injections (1-5 microl) expanded the abdomen in the dorsal-ventral plane, and this expansion was positively correlated with the magnitude of the ensuing diuresis, suggesting the signal for diuretic hormone release originates from stretch receptors in abdominal tergal-sternal muscles. The effects of decapitation, severing the ventral nerve cord within the neck or ligaturing the neck, showed the head was needed to initiate and maintain diuresis, but was neither the source of diuretic hormone nor did it control the discharge of urine from the anus. These findings indicate the head is part of the neural-endocrine pathway between abdominal stretch receptors and sites for diuretic hormone release from the thoracic-abdominal ganglion mass. Evidence is presented for Musdo-K having a hormonal role in the control of diuresis, although other neuropeptides may also be implicated.

  12. Resistance risk assessment of 2 insect development inhibitors, diflubenzuron and cyromazine, for control of the housefly musca-domestica l

    DEFF Research Database (Denmark)

    Keiding, J.; El-Khodary, A.S.; Jespersen, Jørgen B.

    1992-01-01

    pressure is compared with other investigations and discussed. Moderate to high resistance to DFB or CYR can develop in housefly populations if the selection pressure is strong, especially when used as feed-through applications on poultry farms where all feed contains DFB or CYR. If the treatment of fly...

  13. Daily activity of the housefly, Musca domestica, is influenced by temperature independent of 3’UTR period gene splicing

    Czech Academy of Sciences Publication Activity Database

    Bazalová, Olga; Doležel, David

    2017-01-01

    Roč. 7, č. 8 (2017), s. 2637-2649 ISSN 2160-1836 R&D Projects: GA ČR(CZ) GA17-01003S EU Projects: European Commission(XE) 316304 - MODBIOLIN Institutional support: RVO:60077344 Keywords : temperature compensation of circadian rhythms * locomotor activity * transcription Subject RIV: ED - Physiology OBOR OECD: Biology (theoretical, mathematical, thermal, cryobiology, biological rhythm), Evolutionary biology Impact factor: 2.861, year: 2016 http://www.g3journal.org/content/early/2017/06/15/g3.117.042374

  14. Structural basis of hAT transposon end recognition by Hermes, an octameric DNA transposase from Musca domestica.

    Science.gov (United States)

    Hickman, Alison B; Ewis, Hosam E; Li, Xianghong; Knapp, Joshua A; Laver, Thomas; Doss, Anna-Louise; Tolun, Gökhan; Steven, Alasdair C; Grishaev, Alexander; Bax, Ad; Atkinson, Peter W; Craig, Nancy L; Dyda, Fred

    2014-07-17

    Hermes is a member of the hAT transposon superfamily that has active representatives, including McClintock's archetypal Ac mobile genetic element, in many eukaryotic species. The crystal structure of the Hermes transposase-DNA complex reveals that Hermes forms an octameric ring organized as a tetramer of dimers. Although isolated dimers are active in vitro for all the chemical steps of transposition, only octamers are active in vivo. The octamer can provide not only multiple specific DNA-binding domains to recognize repeated subterminal sequences within the transposon ends, which are important for activity, but also multiple nonspecific DNA binding surfaces for target capture. The unusual assembly explains the basis of bipartite DNA recognition at hAT transposon ends, provides a rationale for transposon end asymmetry, and suggests how the avidity provided by multiple sites of interaction could allow a transposase to locate its transposon ends amidst a sea of chromosomal DNA. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. The antibiotic resistome of swine manure is significantly altered by association with the Musca domestica larvae gut microbiome.

    Science.gov (United States)

    Wang, Hang; Sangwan, Naseer; Li, Hong-Yi; Su, Jian-Qiang; Oyang, Wei-Yin; Zhang, Zhi-Jian; Gilbert, Jack A; Zhu, Yong-Guan; Ping, Fan; Zhang, Han-Luo

    2017-01-01

    The overuse of antibiotics as veterinary feed additives is potentially contributing to a significant reservoir of antibiotic resistance in agricultural farmlands via the application of antibiotic-contaminated manure. Vermicomposting of swine manure using housefly larvae is a promising biotechnology for waste reduction and control of antibiotic pollution. To determine how vermicomposting influences antibiotic resistance traits in swine manure, we explored the resistome and associated bacterial community dynamics during larvae gut transit over 6 days of treatment. In total, 94 out of 158 antibiotic resistance genes (ARGs) were significantly attenuated (by 85%), while 23 were significantly enriched (3.9-fold) following vermicomposting. The manure-borne bacterial community showed a decrease in the relative abundance of Bacteroidetes, and an increase in Proteobacteria, specifically Ignatzschineria, following gut transit. ARG attenuation was significantly correlated with changes in microbial community succession, especially reduction in Clostridiales and Bacteroidales. Six genomes were assembled from the manure, vermicompost (final product) and gut samples, including Pseudomonas, Providencia, Enterococcus, Bacteroides and Alcanivorax. Transposon-linked ARGs were more abundant in gut-associated bacteria compared with those from manure and vermicompost. Further, ARG-transposon gene cassettes had a high degree of synteny between metagenomic assemblies from gut and vermicompost samples, highlighting the significant contribution of gut microbiota through horizontal gene transfer to the resistome of vermicompost. In conclusion, the larvae gut microbiome significantly influences manure-borne community succession and the antibiotic resistome during animal manure processing.

  16. Resistance to pyrethroid insecticides in house flies, Musca domestica L., (Diptera: Muscidae) collected from urban areas in Punjab, Pakistan.

    Science.gov (United States)

    Khan, Hafiz Azhar Ali; Akram, Waseem; Fatima, Ammara

    2017-12-01

    House flies are one of the major public health pests in urban settings. People usually use insecticides containing pyrethroids for the management of house flies; however, there is a lack of information on pyrethroid resistance in house flies from urban areas. In the present study, resistance to four pyrethroids (beta-cyfluthrin, deltamethrin, permethrin, transfluthrin) was assessed in house flies collected from urban areas of Punjab, Pakistan. Significant levels of resistance to all the pyrethroids were found in different strains of house flies. The resistance ratios (RRs) at the median lethal dose (LD 50 ) level were in the range of 5.25- to 11.02-fold for beta-cyfluthrin, 7.22- to 19.31-fold for deltamethrin, 5.36- to 16.04-fold for permethrin, and 9.05- to 35.50-fold for transfluthrin. Pairwise comparison of the log LD 50 s revealed a highly significant correlation (p house flies from urban areas of Punjab. Regular resistance monitoring surveys and integrated approaches for the management of house flies are needed to retain the efficacy of these insecticides for a longer period of time.

  17. Evaluation of cimi-shield knock-out bed bug eliminator against house fly (Musca domestica) adults.

    Science.gov (United States)

    Cimi-Shield Knock-Out (CSKO) Bed Bug Eliminator is a green treatment labeled for use against bed bugs, carpet beetles, ants, roaches, fleas, ticks, silverfish, millipedes and centipedes. The active ingredient is soybean oil. If CSKO is formulated according to label instructions and sprayed directly ...

  18. Analysis of differentially expressed genes related to resistance in spinosad- and neonicotinoid-resistant Musca domestica L. (Diptera: Muscidae) strains

    DEFF Research Database (Denmark)

    Castberg, Dorte Heidi Højland; Kristensen, Michael

    2017-01-01

    strains differing significantly in their response to insecticides. High differential expression of P450s and genes coding for cuticle protein indicates a combination of factors involved in metabolic neonicotinoid and spinosad resistance. Conclusion Resistance in these strains is apparently not linked...... interesting in terms of neonicotinoid resistance, while cyp4d9 was overexpressed in 791spin compared to spinosad-susceptible strains. GSTs, ESTs and UGTs were mostly overexpressed, but not to the same degree as P450s. We present a comprehensive and comparative picture of gene expression in three housefly......Background The housefly is a global pest that has developed resistance to most insecticides applied against it. Resistance of the spinosad-resistant strain 791spin and the neonicotinoid-resistant 766b strain is believed to be due to metabolism. We investigate differentially expressed genes...

  19. A whole transcriptomal linkage analysis of gene co-regulation in insecticide resistant house flies, Musca domestica

    DEFF Research Database (Denmark)

    Li, Ming; Reid, William R; Zhang, Lee

    2013-01-01

    autosomes, especially between autosomes 2 and 5, suggesting that signaling transduction cascades controlled by GPCRs, protein kinase/phosphates and proteases may be involved in the regulation of resistance P450 gene regulation. Conclusion Taken together, our findings suggested that not only is insecticide......Background Studies suggest that not only is insecticide resistance conferred via multiple gene up-regulation, but it is mediated through the interaction of regulatory factors. However, no regulatory factors in insecticide resistance have yet been identified, and there has been no examination...... of the regulatory interaction of resistance genes. Our current study generated the first reference transcriptome from the adult house fly and conducted a whole transcriptome analysis for the multiple insecticide resistant strain ALHF (wild-type) and two insecticide susceptible strains: aabys (with morphological...

  20. Evaluation of Commercial and Field-Expedient Baited Traps for House Flies, Musca domestica L. (Diptera: Muscidae)

    Science.gov (United States)

    2009-01-09

    32212, U.S.A. Received 7 September 2008; Accepted 9 January 2009 ABSTRACT: A comparison of nine commercial baited fly traps on Florida dairy farms...baits relied on natural products such as fermented egg slurries (Willson and Mulla 1973) or combinations of such items as molasses, milk, yeast, grain...2006 on four dairy farms in Gilchrist and Alachua counties, FL. Traps were placed near the calf pens on three of the farms and by the commodity

  1. Expression of xenobiotic metabolizing cytochrome P450 genes in a spinosad-resistant Musca domestica L. strain.

    Directory of Open Access Journals (Sweden)

    Dorte H Højland

    Full Text Available Spinosad is important in pest management strategies of multiple insect pests. However, spinosad resistance is emerging in various pest species. Resistance has in some species been associated with alterations of the target-site receptor, but in others P450s seems to be involved. We test the possible importance of nine cytochrome P450 genes in the spinosad-resistant housefly strain 791spin and investigate the influence of spinosad on P450 expression in four other housefly strains.Significant differences in P450 expression of the nine P450 genes in the four strains after spinosad treatment were identified in 40% of cases, most of these as induction. The highly expressed CYP4G2 was induced 6.6-fold in the insecticide susceptible WHO-SRS females, but decreased 2-fold in resistant 791spin males. CYP6G4 was constitutively higher expressed in the resistant strain compared to the susceptible strain. Furthermore, CYP6G4 gene expression was increased in susceptible WHO-SRS flies by spinosad while the expression level did not alter significantly in resistant fly strains. Expression of CYP6A1 and male CYP6D3 was constitutively higher in the resistant strain compared to the susceptible. However, in both cases male expression was higher than female expression.CYP4G2, CYP6A1, CYP6D3 and CYP6G4 have expressions patterns approaching the expectations of a hypothesized sex specific spinosad resistance gene. CYP4G2 fit requirements of a spinosad resistance gene best, making it the most likely candidate. The overall high expression level of CYP4G2 throughout the strains also indicates importance of this gene. However, the data on 791spin are not conclusive concerning spinosad resistance and small contributions from multiple P450s with different enzymatic capabilities could be speculated to do the job in 791spin. Differential expression of P450s between sexes is more a rule than an exception. Noteworthy differences between spinosad influenced expression of P450 genes between a field population and established laboratory strains were shown.

  2. Phytochemical screening and in vitro acetylcholinesterase inhibitory ...

    African Journals Online (AJOL)

    Phytochemical screening and in vitro acetylcholinesterase inhibitory activity of seven plant extracts. Titilayo Johnson, Oduje A. Akinsanmi, Enoch J. Banbilbwa, Tijani A. Yahaya, Karima Abdulaziz, Kolade Omole ...

  3. Synthesis of Novel Chalcones as Acetylcholinesterase Inhibitors

    Directory of Open Access Journals (Sweden)

    Thanh-Dao Tran

    2016-07-01

    Full Text Available A new series of benzylaminochalcone derivatives with different substituents on ring B were synthesized and evaluated as inhibitors of acetylcholinesterase. The study is aimed at identification of novel benzylaminochalcones capable of blocking acetylcholinesterase activity for further development of an approach to Alzheimer’s disease treatment. These compounds were produced in moderate to good yields via Claisen-Schmidt condensation and subjected to an in vitro acetylcholinesterase inhibition assay, using Ellman’s method. The in silico docking procedure was also employed to identify molecular interactions between the chalcone compounds and the enzyme. Compounds with ring B bearing pyridin-4-yl, 4-nitrophenyl, 4-chlorophenyl and 3,4-dimethoxyphenyl moieties were discovered to exhibit significant inhibitory activities against acetylcholinesterase, with IC50 values ranging from 23 to 39 µM. The molecular modeling studies are consistent with the hypothesis that benzylaminochalcones could exert their effects as dual-binding-site acetylcholinesterase inhibitors, which might simultaneously enhance cholinergic neurotransmission and inhibit β-amyloid aggregation through binding to both catalytic and peripheral sites of the enzyme. These derivatives could be further developed to provide novel leads for the discovery of new anti-Alzheimer drugs in the future.

  4. Myrtenal inhibits acetylcholinesterase, a known Alzheimer target.

    Science.gov (United States)

    Kaufmann, Dorothea; Dogra, Anudeep Kaur; Wink, Michael

    2011-10-01

    Inhibition of acetylcholinesterase (AChE) is a common treatment for early stages of the most general form of dementia, Alzheimer's disease. In this study selected components of essential oils, which carry a variety of important functional groups, were tested for their in-vitro anti-acetylcholinesterase activity. In-vitro anti-acetylcholinesterase activity was measured by an adapted version of Ellman's colorimetric assay. 1,8-cineole, carvacrol, myrtenal and verbenone apparently inhibited AChE; the highest inhibitory activity was observed for myrtenal (IC50 = 0.17 mm). This is the first study showing the AChE inhibitory activity of myrtenal. Our investigations provided evidence for the efficacy of monoterpenes as inhibitors of AChE. © 2011 The Authors. JPP © 2011 Royal Pharmaceutical Society.

  5. Acetylcholinesterase and Butyrylcholinesterase Inhibited by Paraoxon

    Czech Academy of Sciences Publication Activity Database

    Kuča, K.; Musilová, L.; Paleček, J.; Církva, Vladimír; Paar, M.; Musílek, K.; Hrabinová, M.; Pohanka, M.; Zdarová Karasová, J.; Jun, D.

    2009-01-01

    Roč. 14, č. 12 (2009), s. 4915-4921 ISSN 1420-3049 Grant - others:MO0(CZ) FZV0000604 Institutional research plan: CEZ:AV0Z40720504 Keywords : acetylcholinesterase * reactivator * oxime Subject RIV: CC - Organic Chemistry Impact factor: 1.738, year: 2009

  6. R. Strauss: Symphonia domestica, Op. 53 / David Nice

    Index Scriptorium Estoniae

    Nice, David

    1994-01-01

    Uuest heliplaadist "R. Strauss: Symphonia domestica, Op. 53. National Youth Orchestra of Great Britain / Christopher Seaman. Pickwick IMP Classics CD PCD 1080; Selected comparisons: SNO, Järvi (3/88) Chandos CHAN 8572"

  7. Differential dependence of apple ( Malus domestica Borkh.) cultivars ...

    African Journals Online (AJOL)

    Golden Delicious' and 'Topred' apple (Malus domestica Borkh.) peel on the xanthophyll cycle for photoprotection was studied under laboratory conditions. Mature fruit peel was either treated with 1 mm dithiothreitol (DTT) to inhibit the ...

  8. A simple radiometric in vitro assay for acetylcholinesterase inhibitors

    International Nuclear Information System (INIS)

    Guilarte, T.R.; Burns, H.D.; Dannals, R.F.; Wagner, H.N. Jr.

    1983-01-01

    A radiometric method for screening acetylcholinesterase inhibitors has been described. The method is based on the production of [ 14 C]carbon dioxide from the hydrolysis of acetylcholine. The inhibitory concentration at 50% (IC50) values for several known acetylcholinesterase inhibitors were in agreement with literature values. The new radiometric method is simple, inexpensive, and has the potential for automation

  9. Acetylcholinesterase Inhibitory and Antioxidant Properties of Euphorbiacharacias Latex

    Directory of Open Access Journals (Sweden)

    Francesca Pintus

    2013-03-01

    Full Text Available The aim of the present study was to evaluate the acetylcholinesterase inhibitory capacity and the antioxidant properties of extracts of Euphorbia characias latex, a Mediterranean shrub. We performed a new extraction method involving the use of the trichloroacetic acid. The extract showed high antioxidant activity, was rich in total polyphenolic and flavonoid content and exhibited substantial inhibition of acetylcholinesterase activity.

  10. Acetyl-cholinesterase Enzyme Inhibitory Effect of Calophyllum species

    African Journals Online (AJOL)

    Purpose: To search for new acetylcholinesterase enzyme inhibitors from Calopyllum species. Methods: Six stem bark extracts of Calophyllum inophyllum, C. soulattri, C. teysmannii, C. lowii, C. benjaminum and C. javanicum were subjected to anti-cholinesterase analysis against acetylcholinesterase (AChE) enzyme using ...

  11. Entomophthora muscae — moisture as a factor affecting its transmission and conidial germination

    Directory of Open Access Journals (Sweden)

    John P. Kramer

    2014-08-01

    Full Text Available The role played by moisture in the transmission of Entomophthora muscae and in the germination of its conidia was investigated. A majority of adult house flies exposed to conidial showers that fell upon surfaces covered with droplets of condensation acquired the parasite, while no flies exposed to conidial showers that fell upon dry surfaces did so. A microscopical study of conidial showers showed that germination was practically non-existent on dry surfaces while a vast majority of conidia that fell upon a droplet-covered surface germinated. A method for the in vivo culture of E. muscae was developpd and 11 serial passages of the fungus were achieved. Resting spores rather than conidia became the dominant form produced in the cadavers, and flies in a twelfth group remained unifected.

  12. The Musca cloud: A 6 pc-long velocity-coherent, sonic filament

    Science.gov (United States)

    Hacar, A.; Kainulainen, J.; Tafalla, M.; Beuther, H.; Alves, J.

    2016-03-01

    Filaments play a central role in the molecular clouds' evolution, but their internal dynamical properties remain poorly characterized. To further explore the physical state of these structures, we have investigated the kinematic properties of the Musca cloud. We have sampled the main axis of this filamentary cloud in 13CO and C18O (2-1) lines using APEX observations. The different line profiles in Musca shows that this cloud presents a continuous and quiescent velocity field along its ~6.5 pc of length. With an internal gas kinematics dominated by thermal motions (I.e. σNT/cs ≲ 1) and large-scale velocity gradients, these results reveal Musca as the longest velocity-coherent, sonic-like object identified so far in the interstellar medium. The transonic properties of Musca present a clear departure from the predicted supersonic velocity dispersions expected in the Larson's velocity dispersion-size relationship, and constitute the first observational evidence of a filament fully decoupled from the turbulent regime over multi-parsec scales. This publication is based on data acquired with the Atacama Pathfinder Experiment (APEX). APEX is a collaboration between the Max-Planck-Institut fuer Radioastronomie, the European Southern Observatory, and the Onsala Space Observatory (ESO programme 087.C-0583).The reduced datacubes as FITS files are only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (ftp://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/587/A97

  13. New Acetylcholinesterase Inhibitors for Alzheimer's Disease

    Directory of Open Access Journals (Sweden)

    Mona Mehta

    2012-01-01

    Full Text Available Acetylcholinesterase (AChE remains a highly viable target for the symptomatic improvement in Alzheimer's disease (AD because cholinergic deficit is a consistent and early finding in AD. The treatment approach of inhibiting peripheral AchE for myasthenia gravis had effectively proven that AchE inhibition was a reachable therapeutic target. Subsequently tacrine, donepezil, rivastigmine, and galantamine were developed and approved for the symptomatic treatment of AD. Since then, multiple cholinesterase inhibitors (ChEI continue to be developed. These include newer ChEIs, naturally derived ChEIs, hybrids, and synthetic analogues. In this paper, we summarize the different types of ChEIs in development and their respective mechanisms of actions. This pharmacological approach continues to be active with many promising compounds.

  14. Changes in acetylcholinesterase activities in the developing and ...

    African Journals Online (AJOL)

    Changes in acetylcholinesterase activities in the developing and aging pig brain and hypophyses. ... International Journal of Agriculture and Rural Development. Journal Home · ABOUT ... Open Access DOWNLOAD FULL TEXT Subscription ...

  15. Accumulation of dioxins and PCB in house fly larvae (musca domestica) reared in poultry manure and used in feed for organic laying hens

    DEFF Research Database (Denmark)

    Nordentoft, Steen; Cederberg, Tommy Licht; Fischer, Christian Holst

    2014-01-01

    higher content of the persistent organic environmental contaminants dioxins and PCB than eggs from conventional hens held indoor in cages. 1,2 The elevated levels of dioxins and PCB are most likely due to the hens picking in soils contaminated by industrial activities, burning of waste, chemical spillage...... etc. As manure from free range hens is expected to have elevated contents of dioxins and PCB, we investigated whether larvae reared in this type of manure accumulate dioxins and PCB, and if feeding organic laying hens with these larvae would increase the levels of dioxins and PCB in the hen eggs...... larvae, poultry manure, compost and compound feed as well as pooled egg samples from each group were analysed for levels of dioxins and PCB. Analytical procedure: after extraction of the sample with a mixture of pentane and acetone (88:12), the extracts were cleaned-up on a multilayer silica column...

  16. Co-up-regulation of three P450 genes in response to permethrin exposure in permethrin resistant house flies, Musca domestica.

    Science.gov (United States)

    Zhu, Fang; Li, Ting; Zhang, Lee; Liu, Nannan

    2008-09-25

    Insects may use various biochemical pathways to enable them to tolerate the lethal action of insecticides. For example, increased cytochrome P450 detoxification is known to play an important role in many insect species. Both constitutively increased expression (overexpression) and induction of P450s are thought to be responsible for increased levels of detoxification of insecticides. However, unlike constitutively overexpressed P450 genes, whose expression association with insecticide resistance has been extensively studied, the induction of P450s is less well characterized in insecticide resistance. The current study focuses on the characterization of individual P450 genes that are induced in response to permethrin treatment in permethrin resistant house flies. The expression of 3 P450 genes, CYP4D4v2, CYP4G2, and CYP6A38, was co-up-regulated by permethrin treatment in permethrin resistant ALHF house flies in a time and dose-dependent manner. Comparison of the deduced protein sequences of these three P450s from resistant ALHF and susceptible aabys and CS house flies revealed identical protein sequences. Genetic linkage analysis located CYP4D4v2 and CYP6A38 on autosome 5, corresponding to the linkage of P450-mediated resistance in ALHF, whereas CYP4G2 was located on autosome 3, where the major insecticide resistance factor(s) for ALHF had been mapped but no P450 genes reported prior to this study. Our study provides the first direct evidence that multiple P450 genes are co-up-regulated in permethrin resistant house flies through the induction mechanism, which increases overall expression levels of P450 genes in resistant house flies. Taken together with the significant induction of CYP4D4v2, CYP4G2, and CYP6A38 expression by permethrin only in permethrin resistant house flies and the correlation of the linkage of the genes with resistance and/or P450-mediated resistance in resistant ALHF house flies, this study sheds new light on the functional importance of P450 genes in response to insecticide treatment, detoxification of insecticides, the adaptation of insects to their environment, and the evolution of insecticide resistance.

  17. Investigating the potential of selected natural compounds to increase the potency of pyrethrum against houseflies Musca domestica (Diptera: Muscidae)

    DEFF Research Database (Denmark)

    Joffe, Tanya; Gunning, Robin V; Allen, Geoff R

    2012-01-01

    oil, grapefruit oil and parsley seed oil, with 59, 50 and 41% mortality respectively, compared with 18% mortality with unsynergised pyrethrum. Against 381zb houseflies, the most effective natural synergists were parsley seed oil and dillapiole oil. Esterase inhibition by the natural compounds and PBO...... in vitro showed no correlation with pyrethrum synergism in vivo, whereas the inhibition of oxidases in vitro more closely correlated with pyrethrum synergism in vivo. CONCLUSION: Dillapiole oil and parsley seed oil showed the greatest potential as pyrethrum synergists. PBO remained the most effective...

  18. Persistence of antibiotic-resistant and -sensitive Proteus mirabilis strains in the digestive tract of the housefly (Musca domestica) and green bottle flies (Calliphoridae).

    Science.gov (United States)

    Wei, Ting; Miyanaga, Kazuhiko; Tanji, Yasunori

    2014-10-01

    Synanthropic flies have been implicated in the rapid dissemination of antibiotic-resistant bacteria and resistance determinants in the biosphere. These flies stably harbor a considerable number of bacteria that exhibit resistance to various antibiotics, but the mechanisms underlying this phenomenon remain unclear. In this study, we investigated the persistence of antibiotic-resistant bacteria in the digestive tract of houseflies and green bottle flies, using Proteus mirabilis as a model microorganism. One resistant strain carried the blaTEM and aphA1 genes, and another carried a plasmid containing qnrD gene. Quantitative PCR and 454 pyrosequencing were used to monitor the relative abundance of the Proteus strains, as well as potential changes in the overall structure of the whole bacterial community incurred by the artificial induction of Proteus cultures. Both antibiotic-resistant and -sensitive P. mirabilis strains persisted in the fly digestive tract for at least 3 days, and there was no significant difference in the relative abundance of resistant and sensitive strains despite the lower growth rate of resistant strains when cultured in vitro. Therefore, conditions in the fly digestive tract may allow resistant strains to survive the competition with sensitive strains in the absence of antibiotic selective pressure. The composition of the fly-associated bacterial community changed over time, but the contribution of the artificially introduced P. mirabilis strains to these changes was not clear. In order to explain these changes, it will be necessary to obtain more information about bacterial interspecies antagonism in the fly digestive tract.

  19. Two Step Synthesis of a Non-symmetric Acetylcholinesterase Reactivator

    Directory of Open Access Journals (Sweden)

    Vit Koleckar

    2007-08-01

    Full Text Available The newly developed and very promising acetylcholinesterase reactivator (E-1- (2-hydroxyiminomethylpyridinium-4-(4-hydroxyiminomethylpyridinium-but-2-ene dibromide was prepared using two different pathways via a two-step synthesis involving the appropriate (E-1-(4-bromobut-2-enyl-2- or 4-hydroxyiminomethyl-pyridinium bromides. Afterwards, purities and yields of the desired product prepared by both routes were compared. Finally, its potency to reactivate several nerve agent-inhibited acetylcholinesterases was tested.

  20. X-Ray Transient in Musca (GRS 1121-68 = GS 1124-683)

    DEFF Research Database (Denmark)

    Lund, Niels; Brandt, Søren; Makino, F.

    1991-01-01

    Research Institute, Lyngby; and Space Research Institute, Moscow), report: "A new x-ray source in Musca, GRS 1121-68, has been discovered in data obtained by the WATCH detectors on Jan. 9. The spectrum of the source was harder than that of the Crab nebula, and the flux was about twice that from t...... was taken as La Silla, showing no new object in the 2 x 2 deg field brighter than mag 7-8; a search plate will be taken with the Schmidt telescope at La Silla....

  1. Polyphenolic Characterization and Antioxidant Activity of Malus domestica and Prunus domestica Cultivars from Costa Rica.

    Science.gov (United States)

    Navarro, Mirtha; Moreira, Ileana; Arnaez, Elizabeth; Quesada, Silvia; Azofeifa, Gabriela; Vargas, Felipe; Alvarado, Diego; Chen, Pei

    2018-01-30

    The phenolic composition of skin and flesh from Malus domestica apples (Anna cultivar) and Prunus domestica plums (satsuma cultivar) commercial cultivars in Costa Rica, was studied using Ultra Performance Liquid Chromatography coupled with High Resolution Mass Spectrometry (UPLC-DAD-ESI-MS) on enriched-phenolic extracts, with particular emphasis in proanthocyanidin and flavonoids characterization. A total of 52 compounds were identified, including 21 proanthocyanidins ([(+)-catechin and (-)-epicatechin]) flavan-3-ols monomers, five procyanidin B-type dimers and two procyanidin A-type dimers, five procyanidin B-type trimers and two procyanidin A-type trimers, as well as one procyanidin B-type tetramer, two procyanidin B-type pentamers, and two flavan-3-ol gallates); 15 flavonoids (kaempferol, quercetin and naringenin derivatives); nine phenolic acids (protochatechuic, caffeoylquinic, and hydroxycinnamic acid derivatives); five hydroxychalcones (phloretin and 3-hydroxyphloretin derivatives); and two isoprenoid glycosides (vomifoliol derivatives). These findings constitute the first report of such a high number and diversity of compounds in skins of one single plum cultivar and of the presence of proanthocyanidin pentamers in apple skins. Also, it is the first time that such a large number of glycosylated flavonoids and proanthocyanidins are reported in skins and flesh of a single plum cultivar. In addition, total phenolic content (TPC) was measured with high values observed for all samples, especially for fruits skins with a TPC of 619.6 and 640.3 mg gallic acid equivalents/g extract respectively for apple and plum. Antioxidant potential using 2,2-diphenyl-1-picrylhidrazyl (DPPH) and oxygen radical absorbance capacity (ORAC) methods were evaluated, with results showing also high values for all samples, especially again for fruit skins with IC 50 of 4.54 and 5.19 µg/mL (DPPH) and 16.8 and 14.6 mmol TE/g (ORAC) respectively for apple and plum, indicating the potential

  2. In silico development of new acetylcholinesterase inhibitors.

    Science.gov (United States)

    Pascoini, A L; Federico, L B; Arêas, A L F; Verde, B A; Freitas, P G; Camps, I

    2018-04-19

    In this work, we made use of fragment-based drug design (FBDD) and de novo design to obtain more powerful acetylcholinesterase (AChE) inhibitors. AChE is associated with Alzheimer's disease (AD). It was found that the cholinergic pathways in the cerebral cortex are compromised in AD and the accompanying cholinergic deficiency contributes to the cognitive deterioration of AD patients. In the FBDD approach, fragments are docked into the active site of the protein. As fragments are molecular groups with a low number of atoms, it is possible to study their interaction with localized amino acids. Once the interactions are measured, the fragments are organized by affinity and then linked together to form new molecules with a high degree of interaction with the active site. In the other approach, we used the de novo design technique starting from reference drugs used in the AD treatment. These drugs were broken into fragments (seeds). In the growing strategy, fragments were added to each seed, growing new molecules. In the linking strategy, two or more separated seeds were linked with different fragments. Both strategies combined produced a library of more than 2 million compounds. This library was filtered using absorption, distribution, metabolism, and excretion properties. The resulting library with around six thousand compounds was filtered again. In this case, structures with Tanimoto coefficients >.85 were discarded. The final library with 1500 compounds was submitted to docking studies. As a result, 10 compounds with better interaction energy than the reference drugs were obtained.

  3. Natural sesquiterpen lactones as acetylcholinesterase inhibitors

    Directory of Open Access Journals (Sweden)

    HOMA HAJIMEHDIPOOR

    2014-06-01

    Full Text Available Background and the purpose of the study: The amount of elder people who suffer from Alzheimer disease is continuously increasing every year. Cholinesterase inhibitors have shown to be effective in alleviating the symptoms of the disease, thus opening a field of research for these treatments. Herbal products, owning a reputation as effective agents in many biological studies are now drawing attention for inhibiting acetylcholinesterase, in other words, Alzheimer disease. In the present study, the ability of three sesquiterpene lactones from Inula oculus-christi and I. aucheriana to inhibit AChE has been evaluated through Ellman assay.Materials and Methods: Gaillardin and pulchellin C were obtained from I. oculus-christi and britannin from I. aucheriana by chromatographic methods. They were dissolved in methanol in concentration of 3 mg/mL and the AChEI activity of the compounds was determined by Ellman method using Acethylthiocholine iodide as the substrate and 5, 5′-dithiobis-2-nitrobenzoic acid as the reagent, in 96-well plates at 405 nm.Results: AChEI activity of the examined compounds was obtained as 67.0, 25.2 and 10.9% in concentration of 300 µg/L for gaillardin, britannin and pulchellin C, respectively.Conclusion: Among the three sesquiterpene lactones, gaillardin with 67% inhibition of AChE could be considered a good candidate for future Alzheimer studies.

  4. A new alkaloid from the fruit of Nandina domestica Thunb.

    Science.gov (United States)

    Peng, Cai-Ying; Liu, Jian-Qun; Zhang, Rui; Shu, Ji-Cheng

    2014-01-01

    A new steroidal alkaloid, (20S,22R,24R)-24-ethyl-3-oxocholest-4-en-22-amino, named as nandsterine (1), together with 10 known alkaloids, palmatine (2), O-methylbulbocapnine (3), nantenine (4), dehydronantenine (5), glaucine (6), didehydroglaucine (7), dehydrocorydaline (8), jatrorrhizine (9), magnoflorine (10) and berberine (11), was isolated from the fruit of Nandina domestica Thunb. Their structures were elucidated by using spectroscopic methods as well as by comparing with the published data. Compound 1 was a new class of steroidal alkaloid isolated from the family Berberidaceae, meanwhile compounds 2, 3, 6-8 and 10 were obtained from N. domestica for the first time. Compound 1 exhibited cytotoxicity against HL-60 cells (human leukaemia) with IC50 values of 52.1 μM.

  5. SERUM ACETYLCHOLINESTERASE LEVEL IN THE PATIENTS OF OPIOID (BROWN SUGAR) DEPENDENCE

    OpenAIRE

    Shah, Nilesh; Dave, Kirti

    1992-01-01

    The authors compared the serum acetylcholinesterase level in the patients of brown sugar dependence and the normal volunteers. Significantly lower level of serum acetylcholinesterase was found in patients of brown sugar dependence.

  6. Altered levels of acetylcholinesterase in Alzheimer plasma.

    Directory of Open Access Journals (Sweden)

    María-Salud García-Ayllón

    Full Text Available BACKGROUND: Many studies have been conducted in an extensive effort to identify alterations in blood cholinesterase levels as a consequence of disease, including the analysis of acetylcholinesterase (AChE in plasma. Conventional assays using selective cholinesterase inhibitors have not been particularly successful as excess amounts of butyrylcholinesterase (BuChE pose a major problem. PRINCIPAL FINDINGS: Here we have estimated the levels of AChE activity in human plasma by first immunoprecipitating BuChE and measuring AChE activity in the immunodepleted plasma. Human plasma AChE activity levels were approximately 20 nmol/min/mL, about 160 times lower than BuChE. The majority of AChE species are the light G(1+G(2 forms and not G(4 tetramers. The levels and pattern of the molecular forms are similar to that observed in individuals with silent BuChE. We have also compared plasma AChE with the enzyme pattern obtained from human liver, red blood cells, cerebrospinal fluid (CSF and brain, by sedimentation analysis, Western blotting and lectin-binding analysis. Finally, a selective increase of AChE activity was detected in plasma from Alzheimer's disease (AD patients compared to age and gender-matched controls. This increase correlates with an increase in the G(1+G(2 forms, the subset of AChE species which are increased in Alzheimer's brain. Western blot analysis demonstrated that a 78 kDa immunoreactive AChE protein band was also increased in Alzheimer's plasma, attributed in part to AChE-T subunits common in brain and CSF. CONCLUSION: Plasma AChE might have potential as an indicator of disease progress and prognosis in AD and warrants further investigation.

  7. Characterization of acetylcholinesterase-inhibition by itopride.

    Science.gov (United States)

    Iwanaga, Y; Kimura, T; Miyashita, N; Morikawa, K; Nagata, O; Itoh, Z; Kondo, Y

    1994-11-01

    Itopride is a gastroprokinetic benzamide derivative. This agent inhibited both electric eel acetylcholinesterase (AChE) and horse serum butyrylcholinesterase (BuChE). The IC50 of itopride with AChE (2.04 +/- 0.27 microM) was, however, 100-fold less than that with BuChE, whereas in the case of neostigmine with AChE (11.3 +/- 3.4 nM), it was 10-fold less. The recovery of AChE activity inhibited by 10(-7) M neostigmine was partial, but that inhibited by up to 3 x 10(-5) M itopride was complete when the reaction mixture was subjected to ultrafiltration. Double reciprocal plots of the experimental data showed that both Km and Vmax were affected by itopride, suggesting that the inhibition is a "mixed" type, although primarily being an uncompetitive one. The inhibitory effect of itopride on cholinesterase (ChE) activity in guinea pig gastrointestine was much weaker than that on pure AChE. However, in the presence of a low dose of diisopropyl fluorophosphate, just enough to inhibit BuChE but not AChE, the IC50s of itopride against ChE activities were found to be about 0.5 microM. In conclusion, itopride exerts reversible and a "mixed" type of inhibition preferably against AChE. The IC50 of itopride for electric eel and guinea pig gastrointestinal AChE inhibition was 200 times and 50 times as large as that of neostigmine, respectively.

  8. Caffeine Inhibits Acetylcholinesterase, But Not Butyrylcholinesterase

    Directory of Open Access Journals (Sweden)

    Petr Dobes

    2013-05-01

    Full Text Available Caffeine is an alkaloid with a stimulant effect in the body. It can interfere in transmissions based on acetylcholine, epinephrine, norepinephrine, serotonin, dopamine and glutamate. Clinical studies indicate that it can be involved in the slowing of Alzheimer disease pathology and some other effects. The effects are not well understood. In the present work, we focused on the question whether caffeine can inhibit acetylcholinesterase (AChE and/or, butyrylcholinesterase (BChE, the two enzymes participating in cholinergic neurotransmission. A standard Ellman test with human AChE and BChE was done for altering concentrations of caffeine. The test was supported by an in silico examination as well. Donepezil and tacrine were used as standards. In compliance with Dixon’s plot, caffeine was proved to be a non-competitive inhibitor of AChE and BChE. However, inhibition of BChE was quite weak, as the inhibition constant, Ki, was 13.9 ± 7.4 mol/L. Inhibition of AChE was more relevant, as Ki was found to be 175 ± 9 µmol/L. The predicted free energy of binding was −6.7 kcal/mol. The proposed binding orientation of caffeine can interact with Trp86, and it can be stabilize by Tyr337 in comparison to the smaller Ala328 in the case of human BChE; thus, it can explain the lower binding affinity of caffeine for BChE with reference to AChE. The biological relevance of the findings is discussed.

  9. Gravitational fragmentation caught in the act: the filamentary Musca molecular cloud

    Science.gov (United States)

    Kainulainen, J.; Hacar, A.; Alves, J.; Beuther, H.; Bouy, H.; Tafalla, M.

    2016-02-01

    Context. Filamentary structures are common in molecular clouds. Explaining how they fragment to dense cores is a missing step in understanding their role in star formation. Aims: We perform a case study of whether low-mass filaments are close to hydrostatic prior to their fragmentation, and whether their fragmentation agrees with gravitational fragmentation models. To accomplish this, we study the ~6.5 pc long Musca molecular cloud, which is an ideal candidate for a filament at an early stage of fragmentation. Methods: We employ dust extinction mapping, in conjunction with near-infrared JHKS-band data from the CTIO/NEWFIRM instrument, and 870 μm dust continuum emission data from the APEX/LABOCA instrument to estimate column densities in Musca. We use the data to identify fragments from the cloud and to determine the radial density distribution of its filamentary part. We compare the cloud's morphology with 13CO and C18O line emission observed with the APEX/SHeFI instrument. Results: The Musca cloud is pronouncedly fragmented at its ends, but harbors a remarkably well-defined, ~1.6 pc long filament in its center region. The line mass of the filament is 21-31 M⊙ pc-1 and the full width at half maximum (FWHM) 0.07 pc. The radial profile of the filament can be fitted with a Plummer profile, which has the power-index of 2.6 ± 11% and is flatter than that of an infinite hydrostatic filament. The profile can also be fitted with a hydrostatic cylinder truncated by external pressure. These models imply a central density of ~5-10 × 104 cm-3. The fragments in the cloud have a mean separation of ~0.4 pc, in agreement with gravitational fragmentation. These properties, together with the subsonic and velocity-coherent nature of the cloud, suggest a scenario in which an initially hydrostatic cloud is currently gravitationally fragmenting. The fragmentation started a few tenths of a Myr ago from the ends of the cloud, leaving its center still relatively nonfragmented

  10. Acetyl-cholinesterase Enzyme Inhibitory Effect of Calophyllum species

    African Journals Online (AJOL)

    1Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, ... Purpose: To search for new acetylcholinesterase enzyme inhibitors from ... Index Medicus, JournalSeek, Journal Citation Reports/Science Edition, .... The data are expressed as mean ± standard .... The authors acknowledge financial support from.

  11. Introducing Dynamic Combinatorial Chemistry: Probing the Substrate Selectivity of Acetylcholinesterase

    Science.gov (United States)

    Angelin, Marcus; Larsson, Rikard; Vongvilai, Pornrapee; Ramstrom, Olof

    2010-01-01

    In this laboratory experiment, college students are introduced to dynamic combinatorial chemistry (DCC) and apply it to determine the substrate selectivity of acetylcholinesterase (AChE). Initially, the students construct a chemical library of dynamically interchanging thioesters and thiols. Then, AChE is added and allowed to select and hydrolyze…

  12. Inhibition of acetylcholinesterase activity by essential oil from Citrus paradisi.

    Science.gov (United States)

    Miyazawa, M; Tougo, H; Ishihara, M

    2001-01-01

    Inhibition of acetylcholinesterase (AChE) activity by essential oils of Citrus paradisi (grapefruit pink in USA) was studied. Inhibition of AChE was measured by the colorimetric method. Nootkatone and auraptene were isolated from C. paradisi oil and showed 17-24% inhibition of AChE activity at the concentration of 1.62 microg/mL.

  13. A eficiência de formação estelar em Musca

    Science.gov (United States)

    Hickel, G. R.; Vilas-Boas, J. W. S.; Roberto, A. J.., Jr.; Khan, R. P.

    2003-08-01

    Apresentamos os resultados de um survey no infravermelho próximo (bandas J e H), executado no Laboratório Nacional de Astrofísica, na direção da nuvem escura em forma de filamento de Musca (observamos ao longo de todo o filamento, além de todas as fontes pontuais IRAS e/ou fontes ROSAT brilhantes a uma distância de 2o da nuvem). Nosso objetivo é determinar a eficiência de formação estelar para esta nuvem escura (massa de estrelas formadas/massa da nuvem), procurando por objetos estelares jovens de pequena massa no seu interior e/ou evoluídos o suficiente para estarem afastados do local de nascimento, mas ainda mostrando características de objetos pré-seqüência principal como emissão de raios-X e excesso de emissão no infravermelho próximo. Este survey não estabeleceu nenhum viés na seleção de fontes pontuais IRAS ou fontes ROSAT, uma vez que a imensa maioria das fontes pontuais IRAS nesta região têm qualidade de fluxo ruim. Os candidatos a objetos estelares jovens foram selecionados pelo excesso no índice de cor (J-H), descontados os efeitos da extinção interestelar na linha de visada, determinada através da emissão estendida no infravermelho distante (IRAS). Estimativas de massa foram feitas para estes candidatos, através da relação massa-luminosidade, para calcular a eficiência de formação estelar de Musca.

  14. Acetylcholinesterase assay for cerebrospinal fluid using bupivacaine to inhibit butyrylcholinesterase

    Directory of Open Access Journals (Sweden)

    Anders Jens

    2001-12-01

    Full Text Available Abstract Background Most test systems for acetylcholinesterase activity (E.C.3.1.1.7. are using toxic inhibitors (BW284c51 and iso-OMPA to distinguish the enzyme from butyrylcholinesterase (E.C.3.1.1.8. which occurs simultaneously in the cerebrospinal fluid. Applying Ellman's colorimetric method, we were looking for a non-toxic inhibitor to restrain butyrylcholinesterase activity. Based on results of previous in vitro studies bupivacaine emerged to be a suitable inhibitor. Results Pharmacokinetic investigations with purified cholinesterases have shown maximum inhibition of butyrylcholinesterase activity and minimal interference with acetylcholinesterase activity at bupivacaine final concentrations between 0.1 and 0.5 mmol/l. Based on detailed analysis of pharmacokinetic data we developed three equations representing enzyme inhibition at bupivacaine concentrations of 0.1, 0.2 and 0.5 mmol/l. These equations allow us to calculate the acetylcholinesterase activity in solutions containing both cholinesterases utilizing the extinction differences measured spectrophotometrically in samples with and without bupivacaine. The accuracy of the bupivacaine-inhibition test could be confirmed by investigations on solutions of both purified cholinesterases and on samples of human cerebrospinal fluid. If butyrylcholinesterase activity has to be assessed simultaneously an independent test using butyrylthiocholine iodide as substrate (final concentration 5 mmol/l has to be conducted. Conclusions The bupivacaine-inhibition test is a reliable method using spectrophotometrical techniques to measure acetylcholinesterase activity in cerebrospinal fluid. It avoids the use of toxic inhibitors for differentiation of acetylcholinesterase from butyrylcholinesterase in fluids containing both enzymes. Our investigations suggest that bupivacaine concentrations of 0.1, 0.2 or 0.5 mmol/l can be applied with the same effect using 1 mmol/l acetylthiocholine iodide as substrate.

  15. Curcumin administration suppress acetylcholinesterase gene expression in cadmium treated rats.

    Science.gov (United States)

    Akinyemi, Ayodele Jacob; Oboh, Ganiyu; Fadaka, Adewale Oluwaseun; Olatunji, Babawale Peter; Akomolafe, Seun

    2017-09-01

    Curcumin, the main polyphenolic component of turmeric (Curcuma longa) rhizomes have been reported to exert anticholinesterase potential with limited information on how they regulate acetylcholinesterase (AChE) gene expression. Hence, this study sought to evaluate the effect of curcumin on cerebral cortex acetylcholinesterase (AChE) activity and their mRNA gene expression level in cadmium (Cd)-treated rats. Furthermore, in vitro effect of different concentrations of curcumin (1-5μg/mL) on rat cerebral cortex AChE activity was assessed. Animals were divided into six groups (n=6): group 1 serve as control (without Cd) and receive saline/vehicle, group 2 receive saline plus curcumin at 25mg/kg, group 3 receive saline plus curcumin 50mg/kg, group 4 receive Cd plus vehicle, group 5 receive Cd plus curcumin at 25mg/kg and group 6 receive Cd plus curcumin at 50mg/kg. Rats received Cd (2.5mg/kg) and curcumin (25 and 50mg/kg, respectively) by oral gavage for 7days. Acetylcholinesterase activity was measured by Ellman's method and AChE expression was carried out by a quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) assay. We observed that acute administration of Cd increased acetylcholinesterase activity and in addition caused a significant (Pcurcumin inhibited AChE activity and alters AChE mRNA levels when compared to Cd-treated group. In addition, curcumin inhibits rat cerebral cortex AChE activity in vitro. In conclusion, curcumin exhibit anti-acetylcholinesterase activity and suppressed AChE mRNA gene expression level in Cd exposed rats, thus providing some biochemical and molecular evidence on the therapeutic effect of this turmeric-derived compound in treating neurological disorders including Alzheimer's disease. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Novel cycloartane-type triterpenoid from the fruits of Nandina domestica.

    Science.gov (United States)

    Kodai, Tetsuya; Horiuchi, Yoshinori; Nishioka, Yasuhiro; Noda, Naoki

    2010-04-01

    A novel cycloartane-type triterpenoid was isolated from the fruits of Nandina domestica (Berberidaceae). The structure was characterized as 24-methylene-3-oxocycloartane 13-carboxylic acid on the basis of NMR spectroscopic data.

  17. Feeding Behavior-Related Toxicity due to Nandina domestica in Cedar Waxwings (Bombycilla cedrorum

    Directory of Open Access Journals (Sweden)

    Moges Woldemeskel

    2010-01-01

    Full Text Available Dozens of Cedar Waxwings were found dead in Thomas County, Georgia, USA, in April 2009. Five of these were examined grossly and microscopically. Grossly, all the examined birds had pulmonary, mediastinal, and tracheal hemorrhages. Microscopically, several tissues and organs were diffusely congested and hemorrhagic. Congestion and hemorrhage were marked in the lungs. Intact and partly digested berries of Nandina domestica Thunb. were the only ingesta found in the gastrointestinal tract of these birds. Due to their voracious feeding behavior, the birds had eaten toxic doses of N. domestica berries. N. domestica contains cyanide and is one of the few berries readily available at this time of the year in the region. The gross and microscopic findings are consistent with lesions associated with cyanide toxicity. This paper for the first time documents toxicity associated with N. domestica in Cedar Waxwings.

  18. Transcriptomic responses to biotic stresses in Malus x domestica: a meta-analysis study

    OpenAIRE

    Balan, Bipin; Marra, Francesco Paolo; Caruso, Tiziano; Martinelli, Federico

    2018-01-01

    RNA-Seq analysis is a strong tool to gain insight into the molecular responses to biotic stresses in plants. The objective of this work is to identify specific and common molecular responses between different transcriptomic data related to fungi, virus and bacteria attacks in Malus x domestica. We analyzed seven transcriptomic datasets in Malus x domestica divided in responses to fungal pathogens, virus (Apple Stem Grooving Virus) and bacteria (Erwinia amylovora). Data were dissected using an...

  19. Internal Diffusion-Controlled Enzyme Reaction: The Acetylcholinesterase Kinetics.

    Science.gov (United States)

    Lee, Sangyun; Kim, Ji-Hyun; Lee, Sangyoub

    2012-02-14

    Acetylcholinesterase is an enzyme with a very high turnover rate; it quenches the neurotransmitter, acetylcholine, at the synapse. We have investigated the kinetics of the enzyme reaction by calculating the diffusion rate of the substrate molecule along an active site channel inside the enzyme from atomic-level molecular dynamics simulations. In contrast to the previous works, we have found that the internal substrate diffusion is the determinant of the acetylcholinesterase kinetics in the low substrate concentration limit. Our estimate of the overall bimolecular reaction rate constant for the enzyme is in good agreement with the experimental data. In addition, the present calculation provides a reasonable explanation for the effects of the ionic strength of solution and the mutation of surface residues of the enzyme. The study suggests that internal diffusion of the substrate could be a key factor in understanding the kinetics of enzymes of similar characteristics.

  20. Buprofezin inhibits acetylcholinesterase activity in B-biotype Bemisia tabaci.

    Science.gov (United States)

    Cottage, Emma L A; Gunning, Robin V

    2006-01-01

    B-biotype Bemisia tabaci is a severe insect pest worldwide in many ornamental, agricultural, and horticultural industries. Control of this insect is hampered by resistance to many acetylcholinesterase (AChE)-inhibiting insecticides, such as organophosphates and carbamates. Consequently, insect growth regulators such as buprofezin, which act by inhibiting chitin synthesis, are being investigated for use against B-biotype B. tabaci in Australia. This study discusses the effects of buprofezin on B. tabaciAChE.

  1. Enzymatic and biochemical characterization of Bungarus sindanus snake venom acetylcholinesterase

    Directory of Open Access Journals (Sweden)

    M Ahmed

    2012-01-01

    Full Text Available This study analyses venom from the elapid krait snake Bungarus sindanus, which contains a high level of acetylcholinesterase (AChE activity. The enzyme showed optimum activity at alkaline pH (8.5 and 45ºC. Krait venom AChE was inhibited by substrate. Inhibition was significantly reduced by using a high ionic strength buffer; low ionic strength buffer (10 mM PO4 pH 7.5 inhibited the enzyme by 1. 5mM AcSCh, while high ionic strength buffer (62 mM PO4 pH 7.5 inhibited it by 1 mM AcSCh. Venom acetylcholinesterase was also found to be thermally stable at 45ºC; it only lost 5% of its activity after incubation at 45ºC for 40 minutes. The Michaelis-Menten constant (Km for acetylthiocholine iodide hydrolysis was found to be 0.068 mM. Krait venom acetylcholinesterase was also inhibited by ZnCl2, CdCl2, and HgCl2 in a concentrationdependent manner. Due to the elevated levels of AChE with high catalytic activity and because it is more stable than any other sources, Bungarus sindanus venom is highly valuable for biochemical studies of this enzyme.

  2. Microbial hydroxylation and glycosidation of oleanolic acid by Circinella muscae and their anti-inflammatory activities.

    Science.gov (United States)

    Yan, Sensen; Lin, Haijun; Huang, Huilian; Yang, Min; Xu, Bohui; Chen, Guangtong

    2018-05-29

    Biotransformation of oleanolic acid (OA) by Circinella muscae AS 3.2695 was investigated. Nine hydroxylated and glycosylated metabolites (1-9) were obtained. Their structures were elucidated as 3β,7β-dihydroxyolean-12-en-28-oic acid (1), 3β,7β,21β-trihydroxyolean-12-en-28-oic acid (2), 3β,7α,21β-trihydroxyolean-12-en- 28-oic acid (3), 3β,7β,15α-trihydroxyolean-12-en-28-oic acid (4), 7β,15α-dihydroxy- 3-oxo-olean-12-en-28-oic acid (5), 7β-hydroxy-3-oxo-olean-12-en-28-oic acid (6), oleanolic acid-28-O-β-D-glucopyranosyl ester (7), 3β,21β-dihydroxyolean-12-en-28- oic acid-28-O-β-D-glucopyranosyl ester (8), and 3β,7β,15α-trihydroxyolean-12-en- 28-oic acid-28-O-β-D-glucopyranosyl ester (9) by spectroscopic analysis. Among them, compounds 4 and 9 were new compounds. In addition, anti-inflammatory activities were assayed and evaluated for the isolated metabolites. Most of the metabolites exhibited significant inhibitory activities on lipopolysaccharides-induced NO production in RAW 264.7 cells.

  3. Análisis de los compuestos volátiles de la ciruela amarilla (Prunus domestica L. ssp. domestica

    Directory of Open Access Journals (Sweden)

    Yineth Ruiz

    2011-01-01

    Full Text Available El aroma de las frutas se debe a los constituyentes volátiles presentes que, aunque se encuentran en muy bajas concentraciones, contribuyen al aroma global en grados muy diversos. Se hace necesario usar técnicas de aislamiento y concentración que garanticen el análisis de una composición química semejante a la de la fruta. Este trabajo tuvo como objetivo el análisis de los compuestos volátiles de la ciruela amarilla (Prunus domestica L. ssp. domestica por el método de evaporación del aroma asistida por solvente (SAFE. Este método utiliza un equipo de destilación conectado a una bomba de alto vacío que ofrece la posibilidad de aislar rápidamente compuestos volátiles sin daño térmico en diferentes matrices alimentarias. La separación e identificación de los compuestos volátiles se realizó por cromatografía de gases-espectrometría de masas (GC-MS. Se identificaron 47 compuestos (6,55 mg/kg de pulpa de fruta, entre ellos 14 alcoholes, 8 aldehídos, 7 ésteres, 5 cetonas, 4 ácidos carboxílicos, 4 hidrocarburos aromáticos, 3 lactonas, un compuesto azufrado y una hidroxicetona; 16 de ellos se informan por primera vez. El acetato de etilo (2,88 mg/kg, etanol (1 mg/kg y ácido octanoico (0,78 mg/kg fueron los constituyentes volátiles mayoritarios de esta variedad de ciruela.

  4. Análisis de los compuestos volátiles de la ciruela amarilla (Prunus domestica L. ssp. domestica

    Directory of Open Access Journals (Sweden)

    Yineth Ruiz

    2011-06-01

    Full Text Available El aroma de las frutas se debe a los constituyentes volátiles presentes que, aunque se encuentran en muy bajas concentraciones, contribuyen al aroma global en grados muy diversos. Se hace necesario usar técnicas de aislamiento y concentración que garanticen el análisis de una composición química semejante a la de la fruta. Este trabajo tuvo como objetivo el análisis de los compuestos volátiles de la ciruela amarilla (Prunus domestica L. ssp.domestica por el método de evaporación del aroma asistida por solvente (SAFE. Este método utiliza un equipo de destilación conectado a una bomba de alto vacío que ofrece la posibilidad de aislar rápidamente compuestos volátiles sin daño térmico en diferentes matrices alimentarias. La separación e identificación de los compuestos volátiles se realizó por cromatografía de gases-espectrometría de masas (GC-MS. Se identificaron 47 compuestos (6,55 mg/kg de pulpa de fruta, entre ellos 14 alcoholes, 8 aldehídos, 7 ésteres, 5 cetonas, 4 ácidos carboxílicos, 4 hidrocarburos aromáticos, 3 lactonas, un compuesto azufrado y una hidroxicetona; 16 de ellos se informan por primera vez. El acetato de etilo (2,88 mg/kg, etanol (1 mg/kg y ácido octanoico (0,78 mg/kg fueron los constituyentes volátiles mayoritarios de esta variedad de ciruela.

  5. Surface display and bioactivity of Bombyx mori acetylcholinesterase on Pichia pastoris

    Science.gov (United States)

    To construct the Pichia pastoris (P. pastoris) cell surface display system of Bombyx mori acetylcholinesterase (BmAChE), the gene for the anchor protein (AGa1) was obtained from Saccharomyces cerevisiae and was fused with the modified Bombyx mori acetylcholinesterase gene (bmace) and transformed int...

  6. Photoreactivation of UV-induced pyrimidine dimers and erythema in the marsupial Monodelphis domestica

    International Nuclear Information System (INIS)

    Ley, R.D.

    1985-01-01

    Post-UV treatment of the gray, short-tailed opossum Monodelphis domestica with photoreactivating light (320-400 nm) suppressed the appearance of UV-induced erythema as evidenced by an increase in the dose of UV required to elicit an erythemal response. Pre-UV exposure to photoreactivating light had no effect on the UV induction of erythema. The dose-response for the photoreversal of pyrimidine dimers in epidermal DNA of M. domestica was similar to that for the photoreactivation of erythema induction. These data not only support the notion that DNA is the primary chromophore involved in the induction of erythema but also identify pyrimidine dimers as the major DNA change responsible for its induction. These results also identify M. domestica as a useful whole-animal system with which to determine the role of pyrimidine dimers in other photobiological responses of mammalian skin

  7. Acetylcholinesterase inhibitory effects of some plants from Rosaceae

    Directory of Open Access Journals (Sweden)

    S. Esmaeili

    2015-10-01

    Full Text Available Background and objectives: Alzheimer's disease (AD is an age dependent disorder. AD is associated with decrease of brain acetylcholine level. Nowadays, one of the methods for progression inhibition of AD is using acetylcholinesterase inhibitors. Rosaceae is a large plant family. Different biological effects of some species of this family have been reported. The aim of the present study was to assess the acetylcholinesterase inhibitory (AChEI activity of the selected plants belonging to Rosaceae family. Methods: AChEI activity of six species from Rosaceae including Cotoneaster nummularia, Cerasus microcarpa, Amygdalus scoparia, Agrimonia eupatoria, Rosa canina and Rosa damascena were evaluated based on Ellman’s method in concentration of 300 µg/mL using total extracts and methanol fractions which were obtained by maceration. Results: The results showed that the total extract and methanol fraction of the aerial parts of A. eupatoria demonstrated significant AChEI activity with 46.5% and 56.2% inhibition of the enzyme, respectively. Conclusion: According to the results of the AChEI activity of the methanol fraction of A. eupatoria, it seems that the polar components of the species such as flavonoids may be responsible for its effectiveness.

  8. Old and new acetylcholinesterase inhibitors for Alzheimer's disease.

    Science.gov (United States)

    Galimberti, Daniela; Scarpini, Elio

    2016-10-01

    To date, pharmacological treatment of Alzheimer's disease (AD) includes Acetylcholinesterase Inhibitors (AChEIs) for mild-to-moderate AD, and memantine for moderate-to-severe AD. AChEIs reversibly inhibit acetylcholinesterase (AChE), thus increasing the availability of acetylcholine in cholinergic synapses, enhancing cholinergic transmission. These drugs provide symptomatic short-term benefits, without clearly counteracting the progression of the disease. On the wake of successful clinical trials which lead to the marketing of AChEIs donepezil, rivastigmine and galantamine, many compounds with AChEI properties have been developed and tested mainly in Phase I-II clinical trials in the last twenty years. Here, we review clinical trials initiated and interrupted, and those ongoing so far. Despite many clinical trials with novel AChEIs have been carried out after the registration of those currently used to treat mild to moderate AD, none so far has been successful in a Phase III trial and marketed. Alzheimer's disease is a complex multifactorial disorder, therefore therapy should likely address not only the cholinergic system but also additional neurotransmitters. Moreover, such treatments should be started in very mild phases of the disease, and preventive strategies addressed in elderly people.

  9. Language impairment in Alzheimer's disease and benefits of acetylcholinesterase inhibitors

    Directory of Open Access Journals (Sweden)

    Ferris SH

    2013-08-01

    Full Text Available Steven H Ferris,1 Martin Farlow21Alzheimer's Disease Center, Comprehensive Center on Brain Aging, New York University Langone Medical Center, New York, NY, 2Department of Neurology, Indiana University School of Medicine, Indianapolis, IN, USAAbstract: Alzheimer's disease is characterized by progressively worsening deficits in several cognitive domains, including language. Language impairment in Alzheimer's disease primarily occurs because of decline in semantic and pragmatic levels of language processing. Given the centrality of language to cognitive function, a number of language-specific scales have been developed to assess language deficits throughout progression of the disease and to evaluate the effects of pharmacotherapy on language function. Trials of acetylcholinesterase inhibitors, used for the treatment of clinical symptoms of Alzheimer's disease, have generally focused on overall cognitive effects. However, in the current report, we review data indicating specific beneficial effects of acetylcholinesterase inhibitors on language abilities in patients with Alzheimer’s disease, with a particular focus on outcomes among patients in the moderate and severe disease stages, during which communication is at risk and preservation is particularly important.Keywords: Alzheimer's disease, donepezil, cognition, language, communication, clinical trials

  10. Acetylcholinesterase inhibition by somes promising Brazilian medicinal plants.

    Science.gov (United States)

    Feitosa, C M; Freitas, R M; Luz, N N N; Bezerra, M Z B; Trevisan, M T S

    2011-08-01

    A microplate assay and a thin-layer chromatography (TLC) "in situ" assay based on the Ellman assay was used to screen for acetylcholinesterase inhibitors from ethyl acetate and methanol extracts of Brazilian medicinal plants of families that, according to the literature, have traditional uses that might be connected with acetylcholinesterase inhibition. Eighteen species belonging to Convolvulaceae, Crassulaceae, Euphorbiaceae, Leguminosae, Malvaceae, Moraceae, Nyctaginaceae and Rutaceae families were tested. The most active plants were Ipomoea asarifolia (IC50 = 0.12 mg/mL), Jatropha curcas (IC50 = 0.25 mg/mL), Jatropha gossypiifolia (IC50 = 0.05 mg/mL), Kalanchoe brasiliensis (IC50 = 0.16 mg/mL) and Senna alata (IC50 = 0.08 mg/mL). The most promising extracts were the Jatropha gossypiifolia and Senna alata species assuming there were compounds with a similar activity to galanthamine, which should contain about 1% of an active compound, or if present at lower levels even more active compounds than galanthamine (IC50 = 0.37 x 10-3 mg/mL) should be present.

  11. Acetylcholinesterase inhibition by somes promising Brazilian medicinal plants

    Directory of Open Access Journals (Sweden)

    CM. Feitosa

    Full Text Available A microplate assay and a thin-layer chromatography (TLC "in situ" assay based on the Ellman assay was used to screen for acetylcholinesterase inhibitors from ethyl acetate and methanol extracts of Brazilian medicinal plants of families that, according to the literature, have traditional uses that might be connected with acetylcholinesterase inhibition. Eighteen species belonging to Convolvulaceae, Crassulaceae, Euphorbiaceae, Leguminosae, Malvaceae, Moraceae, Nyctaginaceae and Rutaceae families were tested. The most active plants were Ipomoea asarifolia (IC50 = 0.12 mg/mL, Jatropha curcas (IC50 = 0.25 mg/mL, Jatropha gossypiifolia (IC50 = 0.05 mg/mL, Kalanchoe brasiliensis (IC50 = 0.16 mg/mL and Senna alata (IC50 = 0.08 mg/mL. The most promising extracts were the Jatropha gossypiifolia and Senna alata species assuming there were compounds with a similar activity to galanthamine, which should contain about 1% of an active compound, or if present at lower levels even more active compounds than galanthamine (IC50 = 0.37 x 10-3 mg/mL should be present.

  12. Antioxidant Activity and Acetylcholinesterase Inhibition of Grape Skin Anthocyanin (GSA

    Directory of Open Access Journals (Sweden)

    Mehnaz Pervin

    2014-07-01

    Full Text Available We aimed to investigate the antioxidant and acetylcholinesterase inhibitory activities of the anthocyanin rich extract of grape skin. Grape skin anthocyanin (GSA neutralized free radicals in different test systems, such as 2,-2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS and 2,2-diphenyl-1-picrylhydrazyl (DPPH assays, to form complexes with Fe2+ preventing 2,2'-azobis(2-amidinopropane dihydrochloride (AAPH-induced erythrocyte hemolysis and oxidative DNA damage. Moreover, GSA decreased reactive oxygen species (ROS generation in isolated mitochondria thus inhibiting 2',-7'-dichlorofluorescin (DCFH oxidation. In an in vivo study, female BALB/c mice were administered GSA, at 12.5, 25, and 50 mg per kg per day orally for 30 consecutive days. Herein, we demonstrate that GSA administration significantly elevated the level of antioxidant enzymes in mice sera, livers, and brains. Furthermore, GSA inhibited acetylcholinesterase (AChE in the in vitro assay with an IC50 value of 363.61 µg/mL. Therefore, GSA could be an excellent source of antioxidants and its inhibition of cholinesterase is of interest with regard to neurodegenerative disorders such as Alzheimer’s disease.

  13. Research Advances and Detection Methodologies for Microbe-Derived Acetylcholinesterase Inhibitors: A Systemic Review

    Directory of Open Access Journals (Sweden)

    Jingqian Su

    2017-01-01

    Full Text Available Acetylcholinesterase inhibitors (AChEIs are an attractive research subject owing to their potential applications in the treatment of neurodegenerative diseases. Fungi and bacteria are major producers of AChEIs. Their active ingredients of fermentation products include alkaloids, terpenoids, phenylpropanoids, and steroids. A variety of in vitro acetylcholinesterase inhibitor assays have been developed and used to measure the activity of acetylcholinesterases, including modified Ellman’s method, thin layer chromatography bioautography, and the combined liquid chromatography-mass spectrometry/modified Ellman’s method. In this review, we provide an overview of the different detection methodologies, the microbe-derived AChEIs, and their producing strains.

  14. High Efficiency Acetylcholinesterase Immobilization on DNA Aptamer Modified Surfaces

    Directory of Open Access Journals (Sweden)

    Orada Chumphukam

    2014-04-01

    Full Text Available We report here the in vitro selection of DNA aptamers for electric eel acetylcholinesterase (AChE. One selected aptamer sequence (R15/19 has a high affinity towards the enzyme (Kd = 157 ± 42 pM. Characterization of the aptamer showed its binding is not affected by low ionic strength (~20 mM, however significant reduction in affinity occurred at high ionic strength (~1.2 M. In addition, this aptamer does not inhibit the catalytic activity of AChE that we exploit through immobilization of the DNA on a streptavidin-coated surface. Subsequent immobilization of AChE by the aptamer results in a 4-fold higher catalytic activity when compared to adsorption directly on to plastic.

  15. Tissue distribution of human acetylcholinesterase and butyrylcholinesterase messenger RNA

    Energy Technology Data Exchange (ETDEWEB)

    Jbilo, O.; Barteles, C.F.; Chatonnet, A.; Toutant, J.P.; Lockridge, O.

    1994-12-31

    Tissue distribution of human acetyicholinesterase and butyryicholinesterase messenger RNA. 1 Cholinesterase inhibitors occur naturally in the calabar bean (eserine), green potatoes (solanine), insect-resistant crab apples, the coca plant (cocaine) and snake venom (fasciculin). There are also synthetic cholinesterase inhibitors, for example man-made insecticides. These inhibitors inactivate acetyicholinesterase and butyrylcholinesterase as well as other targets. From a study of the tissue distribution of acetylcholinesterase and butyrylcholinesterase mRNA by Northern blot analysis, we have found the highest levels of butyrylcholinesterase mRNA in the liver and lungs, tissues known as the principal detoxication sites of the human body. These results indicate that butyrylcholinesterase may be a first line of defense against poisons that are eaten or inhaled.

  16. Interleukin 6 modulates acetylcholinesterase activity of brain neurons

    International Nuclear Information System (INIS)

    Clarencon, D.; Multon, E.; Galonnier, M.; Estrade, M.; Fournier, C.; Mathieu, J.; Mestries, J.C.; Testylier, G.; Fatome, M.

    1995-01-01

    Classically, radiation injuries results in a peripheral inflammatory process, and we have previously observed an early systemic interleukin 6 (IL-6) release following whole-body irradiation. Besides, we have demonstrated an early decrease of rat or primate brain acetylcholinesterase (AChE) activity a gamma exposure. The object of the present study is to find possible IL-6 systemic effects on the brain AChE activity. We show that, though intravenous (i.v.) or intra-cerebro-ventricular (ICV) injection of IL-6 can induce a drop in rat brain AChE activity, this cytokine induces only a slight decrease of the AChE release in cultured brain cells. (author)

  17. Comedication and Treatment Length in Users of Acetylcholinesterase Inhibitors

    Directory of Open Access Journals (Sweden)

    Anne Sverdrup Efjestad

    2017-02-01

    Full Text Available Background/Aims: Reduced clinical effect on cognitive decline in dementia by acetylcholinesterase inhibitors (AChEIs may be due to concurrent use of drugs with anticholinergic properties. The aim was to analyze the incidence of AChEI use and comedication with drugs with anticholinergic properties and other potential unfavorable effects. Methods: A prospective study applying drug use data from the Norwegian Prescription Database. Anticholinergic Drug Scale (ADS scores were used as a measure of overall anticholinergic burden. Results: Patients with high ADS scores were more frequently discontinuing treatment early. Coprescribing of antipsychotics was strongly associated with early discontinuation of AChEI treatment. Conclusion: Coprescribing with potentially unfavorable medications was common. A high ADS score was associated with early discontinuation of treatment.

  18. Acetylcholinesterase and Nissl staining in the same histological section.

    Science.gov (United States)

    Shipley, M T; Ennis, M; Behbehani, M M

    1989-12-18

    Acetylcholinesterase (AChE) enzyme histochemistry and Nissl staining are commonly utilized in neural architectonic studies. However, the opaque reaction deposit produced by the most commonly used AChE histochemical methods is not compatible with satisfactory Nissl staining. As a result, precise correlation of AChE and Nissl staining necessitates time-consuming comparisons of adjacent sections which may have differential shrinkage. Here, we have modified the Koelle-Friedenwald histochemical reaction for AChE by omitting the final intensification steps. The modified reaction yields a non-opaque reaction product that is selectively visualized by darkfield illumination. This non-intensified darkfield AChE (NIDA) reaction allows clear visualization of Nissl staining in the same histological section. This combined AChE-Nissl method greatly facilitates detailed correlation of enzyme and cytoarchitectonic organization.

  19. Effect of fluorocarbons on acetylcholinesterase activity and some counter measures

    Science.gov (United States)

    Young, W.; Parker, J. A.

    1975-01-01

    An isolated vagal sympathetic heart system has been successfully used for the study of the effect of fluorocarbons (FCs) on cardiac performance and in situ enzyme activity. Dichlorodifluoromethane sensitizes this preparation to sympathetic stimulation and to exogenous epinephrine challenge. Partial and complete A-V block and even cardiac arrest have been induced by epinephrine challenge in the FC sensitized heart. Potassium chloride alone restores the rhythmicity but not the normal contractility of the heart in such a situation. Addition of glucose will, however, completely restore the normal function of the heart which is sensitized by dichlorodifluoromethane. The ED 50 values of acetylcholinesterase activity which are used as a measure of relative effectiveness of fluorocarbons are compared with the maximum permissible concentration. Kinetic studies indicate that all the fluorocarbons tested so far are noncompetitive.

  20. Preclinical and first-in-human evaluation of PRX-105, a PEGylated, plant-derived, recombinant human acetylcholinesterase-R

    International Nuclear Information System (INIS)

    Atsmon, Jacob; Brill-Almon, Einat; Nadri-Shay, Carmit; Chertkoff, Raul; Alon, Sari; Shaikevich, Dimitri; Volokhov, Inna; Haim, Kirsten Y.; Bartfeld, Daniel; Shulman, Avidor; Ruderfer, Ilya; Ben-Moshe, Tehila; Shilovitzky, Orit; Soreq, Hermona; Shaaltiel, Yoseph

    2015-01-01

    PRX-105 is a plant-derived recombinant version of the human ‘read-through’ acetylcholinesterase splice variant (AChE-R). Its active site structure is similar to that of the synaptic variant, and it displays the same affinity towards organophosphorus (OP) compounds. As such, PRX-105 may serve as a bio-scavenger for OP pesticides and chemical warfare agents. To assess its potential use in prophylaxis and treatment of OP poisoning we conducted several preliminary tests, reported in this paper. Intravenous (IV) PRX-105 was administered to mice either before or after exposure to an OP toxin. All mice who received an IV dose of 50 nmol/kg PRX-105, 2 min before being exposed to 1.33 × LD 50 and 1.5 × LD 50 of toxin and 10 min after exposure to 1.5 × LD 50 survived. The pharmacokinetic and toxicity profiles of PRX-105 were evaluated in mice and mini-pigs. Following single and multiple IV doses (50 to 200 mg/kg) no deaths occurred and no significant laboratory and histopathological changes were observed. The overall elimination half-life (t ½ ) in mice was 994 (± 173) min. Additionally, a first-in-human study, to assess the safety, tolerability and pharmacokinetics of the compound, was conducted in healthy volunteers. The t ½ in humans was substantially longer than in mice (average 26.7 h). Despite the small number of animals and human subjects who were assessed, the fact that PRX-105 exerts a protective and therapeutic effect following exposure to lethal doses of OP, its favorable safety profile and its relatively long half-life, renders it a promising candidate for treatment and prophylaxis against OP poisoning and warrants further investigation. - Highlights: • PRX-105 is a PEGylated plant-derived recombinant human acetylcholinesterase-R. • PRX-105 is a promising bio-scavenger for organophosphorous toxins at lethal doses. • PRX-105 was shown to protect animals both prophylactically and post-poisoning. • First-in-human study exhibited its safety

  1. Preclinical and first-in-human evaluation of PRX-105, a PEGylated, plant-derived, recombinant human acetylcholinesterase-R

    Energy Technology Data Exchange (ETDEWEB)

    Atsmon, Jacob [Clinical Research Center, Tel Aviv Sourasky Medical Center, Tel Aviv (Israel); Sackler Faculty of Medicine, Tel Aviv University (Israel); Brill-Almon, Einat; Nadri-Shay, Carmit; Chertkoff, Raul; Alon, Sari [Protalix Biotherapeutics, Science Park, Carmiel (Israel); Shaikevich, Dimitri; Volokhov, Inna; Haim, Kirsten Y. [Clinical Research Center, Tel Aviv Sourasky Medical Center, Tel Aviv (Israel); Sackler Faculty of Medicine, Tel Aviv University (Israel); Bartfeld, Daniel [Protalix Biotherapeutics, Science Park, Carmiel (Israel); Shulman, Avidor, E-mail: avidors@protalix.com [Protalix Biotherapeutics, Science Park, Carmiel (Israel); Ruderfer, Ilya; Ben-Moshe, Tehila; Shilovitzky, Orit [Protalix Biotherapeutics, Science Park, Carmiel (Israel); Soreq, Hermona [Department of Biological Chemistry, Institute of Life Sciences, The Hebrew University of Jerusalem (Israel); Shaaltiel, Yoseph [Protalix Biotherapeutics, Science Park, Carmiel (Israel)

    2015-09-15

    PRX-105 is a plant-derived recombinant version of the human ‘read-through’ acetylcholinesterase splice variant (AChE-R). Its active site structure is similar to that of the synaptic variant, and it displays the same affinity towards organophosphorus (OP) compounds. As such, PRX-105 may serve as a bio-scavenger for OP pesticides and chemical warfare agents. To assess its potential use in prophylaxis and treatment of OP poisoning we conducted several preliminary tests, reported in this paper. Intravenous (IV) PRX-105 was administered to mice either before or after exposure to an OP toxin. All mice who received an IV dose of 50 nmol/kg PRX-105, 2 min before being exposed to 1.33 × LD{sub 50} and 1.5 × LD{sub 50} of toxin and 10 min after exposure to 1.5 × LD{sub 50} survived. The pharmacokinetic and toxicity profiles of PRX-105 were evaluated in mice and mini-pigs. Following single and multiple IV doses (50 to 200 mg/kg) no deaths occurred and no significant laboratory and histopathological changes were observed. The overall elimination half-life (t{sub ½}) in mice was 994 (± 173) min. Additionally, a first-in-human study, to assess the safety, tolerability and pharmacokinetics of the compound, was conducted in healthy volunteers. The t{sub ½} in humans was substantially longer than in mice (average 26.7 h). Despite the small number of animals and human subjects who were assessed, the fact that PRX-105 exerts a protective and therapeutic effect following exposure to lethal doses of OP, its favorable safety profile and its relatively long half-life, renders it a promising candidate for treatment and prophylaxis against OP poisoning and warrants further investigation. - Highlights: • PRX-105 is a PEGylated plant-derived recombinant human acetylcholinesterase-R. • PRX-105 is a promising bio-scavenger for organophosphorous toxins at lethal doses. • PRX-105 was shown to protect animals both prophylactically and post-poisoning. • First-in-human study

  2. [Cloning and characterization of Caveolin-1 gene in pigeon, Columba livia domestica].

    Science.gov (United States)

    Zhang, Ying; Yu, Jian-Feng; Yang, Li; Wang, Xing-Guo; Gu, Zhi-Liang

    2010-10-01

    Caveolins, a class of principal proteins forming the structure of caveolae in plasmalemma, were encoded by caveolins gene family. Caveolin-1 gene is a member of caveolins gene family. In the present study, a full-length of 2605 bp caveolin-1 cDNA sequence in Columba livia domestica, which included a 537 bp complete ORF encoding a 178 amino acids long putative peptide, were obtained by using RT-PCR and RACE technique. The Columba livia domestica caveolin-1 CDS shared 80.1% - 93.4% homology with Bos taurus, Canis lupus familiaris, Gallus gallus and Rattus norvegicus. Meanwhile, the putative amino acid sequence of Columba livia domestica caveolin-1 shared 85.4% - 97.2% homology with the above species. The semi-quantity RT-PCR revealed that Caveolin-1 expressions were detectable in all the Columba livia domestica tissues and the expressional level of caveolin-1 gene was high in adipose, medium in various muscles, low in liver. These results demonstrated that Caveolin-1 gene was potentially involved in some metabolic pathways in adipose and muscle.

  3. Modeling Loop Reorganization Free Energies of Acetylcholinesterase: A Comparison of Explicit and Implicit Solvent Models

    National Research Council Canada - National Science Library

    Olson, Mark

    2004-01-01

    ... screening of charge-charge interactions. This paper compares different solvent models applied to the problem of estimating the free-energy difference between two loop conformations in acetylcholinesterase...

  4. EEG SPECTRA, BEHAVIORAL STATES AND MOTOR ACTIVITY IN RATS EXPOSED TO ACETYLCHOLINESTERASE INHIBITOR CHLORPYRIFOS.

    Science.gov (United States)

    Exposure to organophosphate pesticides (OP) has been associated with sleep disorders: insomnia and ?excessive dreaming'. However neuronal mechanisms of these effects have not been analyzed. OP inhibit acetylcholinesterase activity leading to a hyperativity of the brain cholin...

  5. N-[11C]methylpiperidine esters as acetylcholinesterase substrates: an in vivo structure-reactivity study

    International Nuclear Information System (INIS)

    Kilbourn, Michael R.; Nguyen, Thinh B.; Snyder, Scott E.; Sherman, Phillip

    1998-01-01

    A series of simple esters incorporating the N-[ 11 C]methylpiperidine structure were examined as in vivo substrates for acetylcholinesterase in mouse brain. 4-N-[ 11 C]Methylpiperidinyl esters, including the acetate, propionate and isobutyrate esters, are good in vivo substrates for mammalian cholinesterases. Introduction of a methyl group at the 4-position of the 4-piperidinol esters, to form the ester of a teritary alcohol, effectively blocks enzymatic action. Methylation of 4- N-[ 11 C]methylpiperidinyl propionate at the 3-position gives a derivative with increased in vivo reactivity toward acetylcholinesterase. Esters of piperidinecarboxylic acids (nipecotic, isonipecotic and pipecolinic acid ethyl esters) are not hydrolyzed by acetylcholinesterase in vivo, nor do they act as in vivo inhibitors of the enzyme. This study has identified simple methods to both increase and decrease the in vivo reactivity of piperidinyl esters toward acetylcholinesterase

  6. Acetylcholinesterase inhibitory activity of lycopodane-type alkaloids from the Icelandic Lycopodium annotinum ssp. alpestre

    DEFF Research Database (Denmark)

    Halldórsdóttir, Elsa Steinunn; Jaroszewski, Jerzy W; Olafsdottir, Elin Soffia

    2010-01-01

    The aim of this study was to investigate structures and acetylcholinesterase inhibitory activities of lycopodane-type alkaloids isolated from an Icelandic collection of Lycopodium annotinum ssp. alpestre. Ten alkaloids were isolated, including annotinine, annotine, lycodoline, lycoposerramine M...

  7. Triterpenoids from Azorella trifurcata (Gaertn.) Pers and their effect against the enzyme acetylcholinesterase

    Energy Technology Data Exchange (ETDEWEB)

    Areche, Carlos; Cejas, Patricia; Thomas, Pablo; San-Martin, Aurelio [University of Chile, Santiago (Chile). Faculty of Sciences. Dept. of Chemistry], e-mail: aurelio@uchile.cl; Astudillo, Luis; Gutierrez, Margarita [University of Talca, Talca (Chile). Inst. of Chemistry of Natural Resource; Loyola, Luis A. [University of Antofagasta (Chile). Faculty of Basic Sciences. Dept. of Chemistry

    2009-07-01

    The inhibition of the enzyme acetylcholinesterase is considered as a strategy for the treatment of Alzheimer's disease, senile dementia, ataxia, and myasthenia gravis. Three lanostane- and two cycloartane-type triterpenes, together with two mulinane-type diterpenes were isolated from petroleum ether extract of the whole shrub of Azorella trifurcata (Gaertn.) Pers. Their effect on the enzyme acetylcholinesterase was assessed as well. In addition, this is the first report of these triterpenes in the genus Azorella. (author)

  8. Triterpenoids from Azorella trifurcata (Gaertn.) Pers and their effect against the enzyme acetylcholinesterase

    International Nuclear Information System (INIS)

    Areche, Carlos; Cejas, Patricia; Thomas, Pablo; San-Martin, Aurelio; Astudillo, Luis; Gutierrez, Margarita; Loyola, Luis A.

    2009-01-01

    The inhibition of the enzyme acetylcholinesterase is considered as a strategy for the treatment of Alzheimer's disease, senile dementia, ataxia, and myasthenia gravis. Three lanostane- and two cycloartane-type triterpenes, together with two mulinane-type diterpenes were isolated from petroleum ether extract of the whole shrub of Azorella trifurcata (Gaertn.) Pers. Their effect on the enzyme acetylcholinesterase was assessed as well. In addition, this is the first report of these triterpenes in the genus Azorella. (author)

  9. The Effect of Parathion on Red Blood Cell Acetylcholinesterase in the Wistar Rat

    Directory of Open Access Journals (Sweden)

    Naofumi Bunya

    2016-01-01

    Full Text Available Organophosphorus (OP pesticide poisoning is a significant problem worldwide. Research into new antidotes for these acetylcholinesterase inhibitors, and even optimal doses for current therapies, is hindered by a lack of standardized animal models. In this study, we sought to characterize the effects of the OP pesticide parathion on acetylcholinesterase in a Wistar rat model that included comprehensive medical care. Methods. Male Wistar rats were intubated and mechanically ventilated and then poisoned with between 20 mg/kg and 60 mg/kg of intravenous parathion. Upon developing signs of poisoning, the rats were treated with standard critical care, including atropine, pralidoxime chloride, and midazolam, for up to 48 hours. Acetylcholinesterase activity was determined serially for up to 8 days after poisoning. Results. At all doses of parathion, maximal depression of acetylcholinesterase occurred at 3 hours after poisoning. Acetylcholinesterase recovered to nearly 50% of baseline activity by day 4 in the 20 mg/kg cohort and by day 5 in the 40 and 60 mg/kg cohorts. At day 8, most rats’ acetylcholinesterase had recovered to roughly 70% of baseline. These data should be useful in developing rodent models of acute OP pesticide poisoning.

  10. A Comparative Analysis of Perinatal Development of Barrel Cortex in Rat, Mouse and Guinea Pig Using Acetylcholinesterase Histochemistry

    OpenAIRE

    ŞENDEMİR, Erdoğan

    2014-01-01

    The role of acetylcholinesterase (AChE) in the developing neocortex was reexamined by comparing its expression in rats, mice and guinea pigs, following the protocol for acetylcholinesterase histochemistry (described in Sendemir et al., 1996) in order to determine the suitability of the breeding colony at UludaÛ University for use as an animal model. A total of 103 pups as well as two adult animals of each species were used. In the rat pups, acetylcholinesterase-rich patches were d...

  11. Investigating the Antioxidant and Acetylcholinesterase Inhibition Activities of Gossypium herbaceam

    Directory of Open Access Journals (Sweden)

    Haji Akber Aisa

    2013-01-01

    Full Text Available Our previous research showed that standardized extract from the flowers of the Gossypium herbaceam labeled GHE had been used in clinical trials for its beneficial effects on brain functions, particularly in connection with age-related dementia and Alzheimer’s disease (AD. The aim of this work was to determine the components of this herb and the individual constituents of GHE. In order to better understand this herb for AD treatment, we investigated the acetylcholinesterase (AChE inhibition and antioxidant activity of GHE as well as the protective effects to PC12 cells against cytotoxicity induced by tertiary butyl hydroperoxide (tBHP using in vitro assays. The antioxidant activities were assessed by measuring their capabilities for scavenging 1,1-diphenyl-2-picylhydrazyl (DPPH and 2-2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS free radical as well as in inhibiting lipid peroxidation. Our data showed that GHE exhibited certain activities against AChE and also is an efficient free radical scavenger, which may be helpful in preventing or alleviating patients suffering from AD.

  12. Efficacy and safety of Curcuma domestica extracts compared with ibuprofen in patients with knee osteoarthritis: a multicenter study

    Science.gov (United States)

    Kuptniratsaikul, Vilai; Dajpratham, Piyapat; Taechaarpornkul, Wirat; Buntragulpoontawee, Montana; Lukkanapichonchut, Pranee; Chootip, Chirawan; Saengsuwan, Jittima; Tantayakom, Kesthamrong; Laongpech, Supphalak

    2014-01-01

    Objective To determine the efficacy and safety of Curcuma domestica extracts in pain reduction and functional improvement. Methods 367 primary knee osteoarthritis patients with a pain score of 5 or higher were randomized to receive ibuprofen 1,200 mg/day or C. domestica extracts 1,500 mg/day for 4 weeks. The main outcomes were Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) total, WOMAC pain, WOMAC stiffness, and WOMAC function scores. Adverse events (AEs) were also recorded. Results 185 and 182 patients were randomly assigned into C. domestica extracts and ibuprofen groups, respectively. The baseline characteristics were no different between groups. The mean of all WOMAC scores at weeks 0, 2, and 4 showed significant improvement when compared with the baseline in both groups. After using the noninferiority test, the mean difference (95% confidence interval) of WOMAC total, WOMAC pain, and WOMAC function scores at week 4 adjusted by values at week 0 of C. domestica extracts were noninferior to those for the ibuprofen group (P=0.010, P=0.018, and P=0.010, respectively), except for the WOMAC stiffness subscale, which showed a trend toward significance (P=0.060). The number of patients who developed AEs was no different between groups. However, the number of events of abdominal pain/discomfort was significantly higher in the ibuprofen group than that in the C. domestica extracts group (P=0.046). Most subjects (96%–97%) were satisfied with the treatment, and two-thirds rated themselves as improved in a global assessment. Conclusion C. domestica extracts are as effective as ibuprofen for the treatment of knee osteoarthritis. The side effect profile was similar but with fewer gastrointestinal AE reports in the C. domestica extracts group. PMID:24672232

  13. The Occurrence of the Cicada Cicadatra persica on Apple Trees, Malus domestica, in Erneh, Syria

    Science.gov (United States)

    Dardar, Marah A.; Belal, Hamzeh M.R.; Basheer, Abedlnabi M.

    2013-01-01

    An infestation of Cicadatra persica KirKaldy (Hemiptera: Cicadidae) on apple trees, Malus domestica Borkhausen (Rosales: Rosaceae), was reported for the first time in the apple fruit orchards of Erneh, Syria. Nymphs, adults, exuvia, and exit holes in the soil were observed. The species was identified as C. persica based on morphological characters. Some biological observations and an acoustic analysis of the male's songs were also achieved. PMID:23909877

  14. The development of enamel tubules during the formation of enamel in the marsupial Monodelphis domestica.

    OpenAIRE

    Sasagawa, I; Ferguson, M W

    1991-01-01

    In Monodelphis domestica, although both processes from odontoblasts and projections from ameloblasts were found in developing enamel, the majority of the contents of enamel tubules were probably processes that originated from odontoblasts. Processes from odontoblasts penetrating into enamel touched part of the ameloblasts in the stage of enamel formation. No specialised cell junctions were seen at the adherence between the two. There were no enamel tubules in the aprismatic and pseudoprismati...

  15. Acute administration of fenproporex increased acetylcholinesterase activity in brain of young rats.

    Science.gov (United States)

    Teodorak, Brena P; Ferreira, Gabriela K; Scaini, Giselli; Wessler, Letícia B; Heylmann, Alexandra S; Deroza, Pedro; Valvassori, Samira S; Zugno, Alexandra I; Quevedo, João; Streck, Emilio L

    2015-08-01

    Fenproporex is the second most commonly amphetamine-based anorectic consumed worldwide; this drug is rapidly converted into amphetamine, in vivo, and acts by increasing dopamine levels in the synaptic cleft. Considering that fenproporex effects on the central nervous system are still poorly known and that acetylcholinesterase is a regulatory enzyme which is involved in cholinergic synapses and may indirectly modulate the release of dopamine, the present study investigated the effects of acute administration of fenproporex on acetylcholinesterase activity in brain of young rats. Young male Wistar rats received a single injection of fenproporex (6.25, 12.5 or 25mg/kg i.p.) or vehicle (2% Tween 80). Two hours after the injection, the rats were killed by decapitation and the brain was removed for evaluation of acetylcholinesterase activity. Results showed that fenproporex administration increased acetylcholinesterase activity in the hippocampus and posterior cortex, whereas in the prefrontal cortex, striatum and cerebellum the enzyme activity was not altered. In conclusion, in the present study we demonstrated that acute administration of fenproporex exerts an effect in the cholinergic system causing an increase in the activity of acetylcholinesterase in a dose-dependent manner in the hippocampus and posterior cortex. Thus, we suggest that the imbalance in cholinergic homeostasis could be considered as an important pathophysiological mechanism underlying the brain damage observed in patients who use amphetamines such as fenproporex.

  16. Acute administration of fenproporex increased acetylcholinesterase activity in brain of young rats

    Directory of Open Access Journals (Sweden)

    BRENA P. TEODORAK

    2015-08-01

    Full Text Available Fenproporex is the second most commonly amphetamine-based anorectic consumed worldwide; this drug is rapidly converted into amphetamine, in vivo, and acts by increasing dopamine levels in the synaptic cleft. Considering that fenproporex effects on the central nervous system are still poorly known and that acetylcholinesterase is a regulatory enzyme which is involved in cholinergic synapses and may indirectly modulate the release of dopamine, the present study investigated the effects of acute administration of fenproporex on acetylcholinesterase activity in brain of young rats. Young male Wistar rats received a single injection of fenproporex (6.25, 12.5 or 25mg/kg i.p. or vehicle (2% Tween 80. Two hours after the injection, the rats were killed by decapitation and the brain was removed for evaluation of acetylcholinesterase activity. Results showed that fenproporex administration increased acetylcholinesterase activity in the hippocampus and posterior cortex, whereas in the prefrontal cortex, striatum and cerebellum the enzyme activity was not altered. In conclusion, in the present study we demonstrated that acute administration of fenproporex exerts an effect in the cholinergic system causing an increase in the activity of acetylcholinesterase in a dose-dependent manner in the hippocampus and posterior cortex. Thus, we suggest that the imbalance in cholinergic homeostasis could be considered as an important pathophysiological mechanism underlying the brain damage observed in patients who use amphetamines such as fenproporex.

  17. Transcriptomic responses to biotic stresses in Malus x domestica: a meta-analysis study.

    Science.gov (United States)

    Balan, Bipin; Marra, Francesco Paolo; Caruso, Tiziano; Martinelli, Federico

    2018-01-31

    RNA-Seq analysis is a strong tool to gain insight into the molecular responses to biotic stresses in plants. The objective of this work is to identify specific and common molecular responses between different transcriptomic data related to fungi, virus and bacteria attacks in Malus x domestica. We analyzed seven transcriptomic datasets in Malus x domestica divided in responses to fungal pathogens, virus (Apple Stem Grooving Virus) and bacteria (Erwinia amylovora). Data were dissected using an integrated approach of pathway- and gene- set enrichment analysis, Mapman visualization tool, gene ontology analysis and inferred protein-protein interaction network. Our meta-analysis revealed that the bacterial infection enhanced specifically genes involved in sugar alcohol metabolism. Brassinosteroids were upregulated by fungal pathogens while ethylene was highly affected by Erwinia amylovora. Gibberellins and jasmonates were strongly repressed by fungal and viral infections. The protein-protein interaction network highlighted the role of WRKYs in responses to the studied pathogens. In summary, our meta-analysis provides a better understanding of the Malus X domestica transcriptome responses to different biotic stress conditions; we anticipate that these insights will assist in the development of genetic resistance and acute therapeutic strategies. This work would be an example for next meta-analysis works aiming at identifying specific common molecular features linked with biotic stress responses in other specialty crops.

  18. Novel acetylcholinesterase target site for malaria mosquito control.

    Directory of Open Access Journals (Sweden)

    Yuan-Ping Pang

    2006-12-01

    Full Text Available Current anticholinesterase pesticides were developed during World War II and are toxic to mammals because they target a catalytic serine residue of acetylcholinesterases (AChEs in insects and in mammals. A sequence analysis of AChEs from 73 species and a three-dimensional model of a malaria-carrying mosquito (Anopheles gambiae AChE (AgAChE reported here show that C286 and R339 of AgAChE are conserved at the opening of the active site of AChEs in 17 invertebrate and four insect species, respectively. Both residues are absent in the active site of AChEs of human, monkey, dog, cat, cattle, rabbit, rat, and mouse. The 17 invertebrates include house mosquito, Japanese encephalitis mosquito, African malaria mosquito, German cockroach, Florida lancelet, rice leaf beetle, African bollworm, beet armyworm, codling moth, diamondback moth, domestic silkworm, honey bee, oat or wheat aphid, the greenbug, melon or cotton aphid, green peach aphid, and English grain aphid. The four insects are house mosquito, Japanese encephalitis mosquito, African malaria mosquito, and German cockroach. The discovery of the two invertebrate-specific residues enables the development of effective and safer pesticides that target the residues present only in mosquito AChEs rather than the ubiquitous serine residue, thus potentially offering an effective control of mosquito-borne malaria. Anti-AgAChE pesticides can be designed to interact with R339 and subsequently covalently bond to C286. Such pesticides would be toxic to mosquitoes but not to mammals.

  19. Comparative effect of pesticides on brain acetylcholinesterase in tropical fish.

    Science.gov (United States)

    Assis, Caio Rodrigo Dias; Linhares, Amanda Guedes; Oliveira, Vagne Melo; França, Renata Cristina Penha; Carvalho, Elba Veronica Matoso Maciel; Bezerra, Ranilson Souza; de Carvalho, Luiz Bezerra

    2012-12-15

    Monitoring of pesticides based on acetylcholinesterase (AChE; EC 3.1.1.7) inhibition in vitro avoids interference of detoxification defenses and bioactivation of some of those compounds in non-target tissues. Moreover, environmental temperature, age and stress are able to affect specific enzyme activities when performing in vivo studies. Few comparative studies have investigated the inter-specific differences in AChE activity in fish. Screening studies allow choosing the suitable species as source of AChE to detect pesticides in a given situation. Brain AChE from the tropical fish: pirarucu (Arapaima gigas), cobia (Rachycentron canadum) and Nile tilapia (Oreochromis niloticus) were characterized and their activities were assayed in the presence of pesticides (the organophosphates: dichlorvos, diazinon, chlorpyrifos, temephos, tetraethyl pyrophosphate- TEPP and the carbamates: carbaryl and carbofuran). Inhibition parameters (IC₅₀ and Ki) for each species were found and compared with commercial AChE from electric eel (Electrophorus electricus). Optimal pH and temperature were found to be 8.0 and 35-45 °C, respectively. A. gigas AChE retained 81% of the activity after incubation at 50 °C for 30 min. The electric eel enzyme was more sensitive to the compounds (mainly carbofuran, IC₅₀ of 5 nM), excepting the one from A. gigas (IC₅₀ of 9 nM) under TEPP inhibition. These results show comparable sensitivity between purified and non-purified enzymes suggesting them as biomarkers for organophosphorus and carbamate detection in routine environmental and food monitoring programs for pesticides. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Bivalent ligands derived from Huperzine A as acetylcholinesterase inhibitors.

    Science.gov (United States)

    Haviv, H; Wong, D M; Silman, I; Sussman, J L

    2007-01-01

    The naturally occurring alkaloid Huperzine A (HupA) is an acetylcholinesterase (AChE) inhibitor that has been used for centuries as a Chinese folk medicine in the context of its source plant Huperzia Serrata. The potency and relative safety of HupA rendered it a promising drug for the ameliorative treatment of Alzheimer's disease (AD) vis-à-vis the "cholinergic hypothesis" that attributes the cognitive decrements associated with AD to acetylcholine deficiency in the brain. However, recent evidence supports a neuroprotective role for HupA, suggesting that it could act as more than a mere palliative. Biochemical and crystallographic studies of AChE revealed two potential binding sites in the active-site gorge of AChE, one of which, the "peripheral anionic site" at the mouth of the gorge, was implicated in promoting aggregation of the beta amyloid (Abeta) peptide responsible for the neurodegenerative process in AD. This feature of AChE facilitated the development of dual-site binding HupA-based bivalent ligands, in hopes of concomitantly increasing AChE inhibition potency by utilizing the "chelate effect", and protecting neurons from Abeta toxicity. Crystal structures of AChE allowed detailed modeling and docking studies that were instrumental in enhancing the understanding of underlying principles of bivalent inhibitor-enzyme dynamics. This monograph reviews two categories of HupA-based bivalent ligands, in which HupA and HupA fragments serve as building blocks, with a focus on the recently solved crystallographic structures of Torpedo californica AChE in complex with such bifunctional agents. The advantages and drawbacks of such structured-based drug design, as well as species differences, are highlighted and discussed.

  1. Immobilization of Acetylcholinesterase on Screen-Printed Electrodes. Application to the Determination of Arsenic(III

    Directory of Open Access Journals (Sweden)

    M. Julia Arcos-Martínez

    2010-03-01

    Full Text Available Enzymatic amperometric procedures for measuring arsenic, based on the inhibitive action of this metal on acetylcholinesterase enzyme activity, have been developed. Screen-printed carbon electrodes (SPCEs were used with acetylcholinesterase covalently bonded directly to its surface. The amperometric response of acetylcholinesterase was affected by the presence of arsenic ions, which caused a decrease in the current intensity. The experimental optimum working conditions of pH, substrate concentration and potential applied, were established. Under these conditions, repeatability and reproducibility of biosensors were determined, reaching values below 4% in terms of relative standard deviation. The detection limit obtained for arsenic was 1.1 × 10−8 M for Ach/SPCE biosensor. Analysis of the possible effect of the presence of foreign ions in the solution was performed. The method was applied to determine levels of arsenic in spiked tap water samples.

  2. A novel biosensor method for surfactant determination based on acetylcholinesterase inhibition

    International Nuclear Information System (INIS)

    Kucherenko, I S; Soldatkin, O O; Arkhypova, V M; Dzyadevych, S V; Soldatkin, A P

    2012-01-01

    A novel enzyme biosensor based on acetylcholinesterase inhibition for the determination of surfactants in aqueous solutions is described. Acetylcholinesterase-based bioselective element was deposited via glutaraldehyde on the surface of conductometric transducers. Different variants of inhibitory analysis of surfactants were tested, and finally surfactant's concentration was evaluated by measuring initial rate of acetylcholinesterase inhibition. Besides, we studied the effect of solution characteristics on working parameters of the biosensor for direct measurement of acetylcholine and for inhibitory determination of surfactants. The biosensor's sensitivity to anionic and cationic surfactants (0.35 mg l −1 ) was tested. The high operational stability of the biosensor during determination of acetylcholine (RSD 2%) and surfactants (RSD 11%) was shown. Finally, we discussed the selectivity of the biosensor toward surfactants and other AChE inhibitors. The proposed biosensor can be used as a component of the multibiosensor for ecological monitoring of toxicants. (paper)

  3. Evaluation of acetylcholinesterase inhibitory activity of Brazilian red macroalgae organic extracts

    Directory of Open Access Journals (Sweden)

    Levi P. Machado

    Full Text Available Abstract Alzheimer's disease affects nearly 36.5 million people worldwide, and acetylcholinesterase inhibition is currently considered the main therapeutic strategy against it. Seaweed biodiversity in Brazil represents one of the most important sources of biologically active compounds for applications in phytotherapy. Accordingly, this study aimed to carry out a quantitative and qualitative assessment of Hypnea musciformis (Wulfen J.V. Lamouroux, Ochtodes secundiramea (Montagne M.A. Howe, and Pterocladiella capillacea (S.G. Gmelin Santelices & Hommersand (Rhodophyta in order to determine the AChE effects from their extracts. As a matter of fact, the O. secundiramea extract showed 48% acetylcholinesterase inhibition at 400 μg/ml. The chemical composition of the bioactive fraction was determined by gas chromatography–mass spectrometry (GC–MS; this fraction is solely composed of halogenated monoterpenes, therefore allowing assignment of acetylcholinesterase inhibition activity to them.

  4. Novel coumarin derivatives bearing N-benzyl pyridinium moiety: potent and dual binding site acetylcholinesterase inhibitors.

    Science.gov (United States)

    Alipour, Masoumeh; Khoobi, Mehdi; Foroumadi, Alireza; Nadri, Hamid; Moradi, Alireza; Sakhteman, Amirhossein; Ghandi, Mehdi; Shafiee, Abbas

    2012-12-15

    A novel series of coumarin derivatives linked to benzyl pyridinium group were synthesized and biologically evaluated as inhibitors of both acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). The enzyme inhibitory activity of synthesized compounds was measured using colorimetric Ellman's method. It was revealed that compounds 3e, 3h, 3l, 3r and 3s have shown higher activity compared with donepezil hydrochloride as standard drug. Most of the compounds in these series had nanomolar range IC(50) in which compound 3r (IC(50) = 0.11 nM) was the most active compound against acetylcholinesterase enzyme. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. Phenolic Lipids Affect the Activity and Conformation of Acetylcholinesterase from Electrophorus electricus (Electric eel)

    Science.gov (United States)

    Stasiuk, Maria; Janiszewska, Alicja; Kozubek, Arkadiusz

    2014-01-01

    Phenolic lipids were isolated from rye grains, cashew nutshell liquid (CNSL) from Anacardium occidentale, and fruit bodies of Merrulius tremellosus, and their effects on the electric eel acetylcholinesterase activity and conformation were studied. The observed effect distinctly depended on the chemical structure of the phenolic lipids that were available for interaction with the enzyme. All of the tested compounds reduced the activity of acetylcholinesterase. The degree of inhibition varied, showing a correlation with changes in the conformation of the enzyme tested by the intrinsic fluorescence of the Trp residues of the protein. PMID:24787269

  6. Chagas disease: modulation of the inflammatory response by acetylcholinesterase in hematological cells and brain tissue.

    Science.gov (United States)

    Silva, Aniélen D; Bottari, Nathieli B; do Carmo, Guilherme M; Baldissera, Matheus D; Souza, Carine F; Machado, Vanessa S; Morsch, Vera M; Schetinger, Maria Rosa C; Mendes, Ricardo E; Monteiro, Silvia G; Da Silva, Aleksandro S

    2018-01-01

    Chagas disease is an acute or chronic illness that causes severe inflammatory response, and consequently, it may activate the inflammatory cholinergic pathway, which is regulated by cholinesterases, including the acetylcholinesterase. This enzyme is responsible for the regulation of acetylcholine levels, an anti-inflammatory molecule linked to the inflammatory response during parasitic diseases. Thus, the aim of this study was to investigate whether Trypanosoma cruzi infection can alter the activity of acetylcholinesterase and acetylcholine levels in mice, and whether these alterations are linked to the inflammatory cholinergic signaling pathway. Twenty-four mice were divided into two groups: uninfected (control group, n = 12) and infected by T. cruzi, Y strain (n = 12). The animals developed acute disease with a peak of parasitemia on day 7 post-infection (PI). Blood, lymphocytes, and brain were analyzed on days 6 and 12 post-infection. In the brain, acetylcholine and nitric oxide levels, myeloperoxidase activity, and histopathology were analyzed. In total blood and brain, acetylcholinesterase activity decreased at both times. On the other hand, acetylcholinesterase activity in lymphocytes increased on day 6 PI compared with the control group. Infection by T. cruzi increased acetylcholine and nitric oxide levels and histopathological damage in the brain of mice associated to increased myeloperoxidase activity. Therefore, an intense inflammatory response in mice with acute Chagas disease in the central nervous system caused an anti-inflammatory response by the activation of the cholinergic inflammatory pathway.

  7. Screening of acetylcholinesterase inhibitors in snake venom by electrospray mass spectrometry

    NARCIS (Netherlands)

    Liesener, A.; Perchuc, Anna-Maria; Schöni, Reto; Schebb, Nils Helge; Wilmer, Marianne; Karst, U.

    2007-01-01

    An electrospray ionization/mass spectrometry (ESI/MS)-based assay for the determination of acetylcholinesterase (AChE)-inhibiting activity in snake venom was developed. It allows the direct monitoring of the natural AChE substrate acetylcholine (AC) and the respective product choline. The assay

  8. Evaluating the antioxidant and acetylcholinesterase inhibitory activity of three Centaurea species

    Directory of Open Access Journals (Sweden)

    H. Hajimehdipoor

    2014-01-01

    Full Text Available Factors such as oxidative stress and reduced acetylcholine level have been implicated in Alzheimer’s disease (AD pathology and recently there has been a trend towards natural product research to find potential sources of antioxidants and acetylcholinesterase inhibitors in the plants kingdom. Centaurea is a genus with about 500 species world wild, many of them have shown to possess biologic activity; Centaurea albonites, C. aucheri and C. pseudoscabiosa are three species which little investigation has been carried out about their biological properties. In the present study, the antioxidant and acetylcholinesterase inhibitory activity of the above mentioned species have been evaluated. The ability of the total extract and methanol fraction of the plants to scavenge free radicals has been assessed through DPPH radical scavenging assay, and the acetylcholinesterase inhibitory property has been evaluated by Ellman method. The total extract of all species exhibited moderate antioxidant activity whereas the extracts of C. pseudoscabiosa showed the strongest antioxidant property; its total extract also demonstrated the highest acetylcholinesterase inhibitory activity among the evaluated samples (19.2% inhibition. The results suggest the species as potential sources of natural antioxidants which could be focused in future studies of Alzheimer’s disease.

  9. Dual Binding Site and Selective Acetylcholinesterase Inhibitors Derived from Integrated Pharmacophore Models and Sequential Virtual Screening

    Directory of Open Access Journals (Sweden)

    Shikhar Gupta

    2014-01-01

    Full Text Available In this study, we have employed in silico methodology combining double pharmacophore based screening, molecular docking, and ADME/T filtering to identify dual binding site acetylcholinesterase inhibitors that can preferentially inhibit acetylcholinesterase and simultaneously inhibit the butyrylcholinesterase also but in the lesser extent than acetylcholinesterase. 3D-pharmacophore models of AChE and BuChE enzyme inhibitors have been developed from xanthostigmine derivatives through HypoGen and validated using test set, Fischer’s randomization technique. The best acetylcholinesterase and butyrylcholinesterase inhibitors pharmacophore hypotheses Hypo1_A and Hypo1_B, with high correlation coefficient of 0.96 and 0.94, respectively, were used as 3D query for screening the Zinc database. The screened hits were then subjected to the ADME/T and molecular docking study to prioritise the compounds. Finally, 18 compounds were identified as potential leads against AChE enzyme, showing good predicted activities and promising ADME/T properties.

  10. Inactivation of acetylcholinesterase by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride.

    Science.gov (United States)

    Zang, Lun-Yi; Misra, Hara P

    2003-12-01

    The neurotoxicant 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) has been shown to reversibly inhibit the activity of acetylcholinesterase. The inactivation of the enzyme was detected by monitoring the accumulation of yellow color produced from the reaction between thiocholine and dithiobisnitrobenzoate ion. The kinetic parameter, Km for the substrate (acetylthiocholine), was found to be 0.216 mM and Ki for MPTP inactivation of acetylcholinesterase was found to be 2.14 mM. The inactivation of enzyme by MPTP was found to be dose-dependent. It was found that MPTP is neither a substrate of AChE nor the time-dependent inactivator. The studies of reaction kinetics indicate the inactivation of AChE to be a linear mixed-type inhibition. The dilution assays indicate that MPTP is a reversible inhibitor for AChE. These data suggest that once MPTP enters the basal ganglia of the brain, it can inactivate the acetylcholinesterase enzyme and thereby increase the acetylcholine level in the basal ganglia of brain, leading to potential cell dysfunction. It appears that the nigrostriatal toxicity by MPTP leading to Parkinson's disease-like syndrome may, in part, be mediated via the acetylcholinesterase inactivation.

  11. Stabilizing Acetylcholinesterase on Carbon Electrodes Using Peptide Nanotubes to Produce Effective Biosensors

    Science.gov (United States)

    2012-03-22

    disposable, inhibition 1. Introduction Organophosphates (OPs) are acetyl cholinesterase (AChE) inhibitors with a broad range of potency and toxicity...5 Cholinesterase Biosensors...BIOSENSORS I. Introduction Background Organophosphates (OPs) are acetylcholinesterase (AChE) inhibitors with a broad range of potency and

  12. Antimicrobial activity and acetylcholinesterase inhibition by extracts from chromatin modulated fungi

    Directory of Open Access Journals (Sweden)

    Matheus Thomaz Nogueira Silva Lima

    Full Text Available ABSTRACT Major health challenges as the increasing number of cases of infections by antibiotic multiresistant microorganisms and cases of Alzheimer's disease have led to searching new control drugs. The present study aims to verify a new way of obtaining bioactive extracts from filamentous fungi with potential antimicrobial and acetylcholinesterase inhibitory activities, using epigenetic modulation to promote the expression of genes commonly silenced. For such finality, five filamentous fungal species (Talaromyces funiculosus, Talaromyces islandicus, Talaromyces minioluteus, Talaromyces pinophilus, Penicillium janthinellum were grown or not with DNA methyltransferases inhibitors (procainamide or hydralazine and/or a histone deacetylase inhibitor (suberohydroxamic acid. Extracts from T. islandicus cultured or not with hydralazine inhibited Listeria monocytogenes growth in 57.66 ± 5.98% and 15.38 ± 1.99%, respectively. Increment in inhibition of acetylcholinesterase activity was observed for the extract from P. janthinellum grown with procainamide (100%, when compared to the control extract (39.62 ± 3.76%. Similarly, inhibition of acetylcholinesterase activity increased from 20.91 ± 3.90% (control to 92.20 ± 3.72% when the tested extract was obtained from T. pinophilus under a combination of suberohydroxamic acid and procainamide. Concluding, increases in antimicrobial activity and acetylcholinesterase inhibition were observed when fungal extracts in the presence of DNA methyltransferases and/or histone deacetylase modulators were tested.

  13. Two Bombyx mori acetylcholinesterase genes influence motor control and development in different ways

    Science.gov (United States)

    Among its other biological roles, acetylcholinesterase (AChE, EC 3.1.1.7), encoded by two ace genes in most insects, catalyses the breakdown of acetylcholine, thereby terminating synaptic transmission. ace1 encodes the synaptic enzyme and ace2 has other essential actions in many insect species, such...

  14. Bifunctional phenolic-choline conjugates as anti-oxidants and acetylcholinesterase inhibitors

    Czech Academy of Sciences Publication Activity Database

    Šebestík, Jaroslav; Marques, S. M.; Falé, P. L.; Santos, S.; Arduíno, D. M.; Cardoso, S. M.; Oliveira, S. M.; Serralheiro, M. L. M.; Santos, A. M.

    Roč. 26, č. 4 (485), s. 497 ISSN 1475-6366 Institutional research plan: CEZ:AV0Z40550506 Keywords : acetylcholinesterase inhibitors * antioxidants * hybrid ligands * anti-neurodegeneratives * Alzheimer´s disease Subject RIV: CC - Organic Chemistry

  15. The interaction of quaternary reversible acetylcholinesterase inhibitors with the nicotinic receptor

    Czech Academy of Sciences Publication Activity Database

    Šepsová, V.; Krůšek, Jan; Zdarová Karasová, J.; Zemek, F.; Musílek, K.; Kuča, K.; Soukup, O.

    2014-01-01

    Roč. 63, č. 6 (2014), s. 771-777 ISSN 0862-8408 R&D Projects: GA ČR(CZ) GBP304/12/G069 Institutional support: RVO:67985823 Keywords : acetylcholinesterase inhibitor * nicotin receptor Subject RIV: ED - Physiology Impact factor: 1.293, year: 2014

  16. Fenugreek hydrogel–agarose composite entrapped gold nanoparticles for acetylcholinesterase based biosensor for carbamates detection

    Energy Technology Data Exchange (ETDEWEB)

    Kestwal, Rakesh Mohan; Bagal-Kestwal, Dipali; Chiang, Been-Huang, E-mail: bhchiang@ntu.edu.tw

    2015-07-30

    A biosensor was fabricated to detect pesticides in food samples. Acetylcholinesterase was immobilized in a novel fenugreek hydrogel–agarose matrix with gold nanoparticles. Transparent thin films with superior mechanical strength and stability were obtained with 2% fenugreek hydrogel and 2% agarose. Immobilization of acetylcholinesterase on the membrane resulted in high enzyme retention efficiency (92%) and a significantly prolonged shelf life of the enzyme (half-life, 55 days). Transmission electron microscopy revealed that, gold nanoparticles (10–20 nm in diameter) were uniformly dispersed in the fenugreek hydrogel–agarose–acetylcholinesterase membrane. This immobilized enzyme-gold nanoparticle dip-strip system detected various carbamates, including carbofuran, oxamyl, methomyl, and carbaryl, with limits of detection of 2, 21, 113, and 236 nM (S/N = 3), respectively. Furthermore, the fabricated biosensor exhibited good testing capabilities when used to detect carbamates added to various fruit and vegetable samples. - Highlights: • Acetylcholinesterase (AChE) dip-strip biosensor fabricated to detect carbamates. • AChE entrapped in fenugreek hydrogel–agarose matrix with gold nanoparticles (GNPs). • High enzyme retention efficiency (92%) and shelf life (half-life, 55 days). • Detection limits of carbofuran, oxamyl and methomyl: 2, 21 and 113 nM. • The biosensor had good testing capabilities to detect carbamates in food samples.

  17. Screening for acetylcholinesterase inhibitory activity in cyanobacteria of the genus Nostoc

    Czech Academy of Sciences Publication Activity Database

    Zelík, Petr; Lukešová, Alena; Voloshko, L. N.; Štys, D.; Kopecký, Jiří

    2009-01-01

    Roč. 24, č. 2 (2009), s. 531-536 ISSN 1475-6366 R&D Projects: GA MŠk ME 874 Institutional research plan: CEZ:AV0Z50200510; CEZ:AV0Z60660521 Keywords : acetylcholinesterase * bioactivity * inhibitors Subject RIV: EE - Microbiology, Virology Impact factor: 1.496, year: 2009

  18. Formulation and quality control of Prunus domestica syrup, prepared according to Iranian Traditional Medicine

    Directory of Open Access Journals (Sweden)

    M. Hamzeloo-Moghadam

    2015-05-01

    Full Text Available Background and objectives: Prunus domestica (plum has been considered as a useful remedy for several disorders in Iranian Traditional Medicine (ITM. It has cold and wet temperament and is used as syrup for hot temperament diseases such as hot headache and stomach disorders. In the present study, plum syrup has been formulated according to ITM manuscripts and quality control evaluations have been accomplished to present a suitable formulation. Methods: The fruits of Prunus domestica L. were macerated in water, then decocted. The mixture was filtered. The filtrate was concentrated to have a suitable viscosity. The extract was sweetened by adding sugar (1:2 and heated till sugar was completely dissolved. The final product was evaluated physicochemically and microbiologically according to standard protocols and total phenolics content of the syrup stability was determined. The syrup was assessed in accelerated condition (40 ºC during 6 months. Results: The prepared formulation was a viscose and brown syrup with plum flavor and fragrance. No precipitation and cap locking were observed in the syrup. Dry residue, pH, density, viscosity and total phenolics of the syrup were found 43.1%, 3.49, 1.27 g/ml, 6.5 cP and 152.3 mg/100ml, respectively. No microbial growth was observed in the formulation. In the accelerated stability tests, no remarkable changes were seen in the product. Total phenolics content was decreased 2.2% during 6 months in 40 ºC. Conclusion: The formulated Prunus domestica syrup could be introduced for further mass production after completing the final required evaluations.

  19. Concentration-dependent interactions of the organophosphates chlorpyrifos oxon and methyl paraoxon with human recombinant acetylcholinesterase

    International Nuclear Information System (INIS)

    Kaushik, R.; Rosenfeld, Clint A.; Sultatos, L.G.

    2007-01-01

    For many decades it has been thought that oxygen analogs (oxons) of organophosphorus insecticides phosphorylate the catalytic site of acetylcholinesterase by a mechanism that follows simple Michaelis-Menten kinetics. More recently, the interactions of at least some oxons have been shown to be far more complex and likely involve binding of oxons to a second site on acetylcholinesterase that modulates the inhibitory capacity of other oxon molecules at the catalytic site. The current study has investigated the interactions of chlorpyrifos oxon and methyl paraoxon with human recombinant acetylcholinesterase. Both chlorpyrifos oxon and methyl paraoxon were found to have k i 's that change as a function of oxon concentration. Furthermore, 10 nM chlorpyrifos oxon resulted in a transient increase in acetylthiocholine hydrolysis, followed by inhibition. Moreover, in the presence of 100 nM chlorpyrifos oxon, acetylthiocholine was found to influence both the K d (binding affinity) and k 2 (phosphorylation constant) of this oxon. Collectively, these results demonstrate that the interactions of chlorpyrifos oxon and methyl paraoxon with acetylcholinesterase cannot be described by simple Michaelis-Menten kinetics but instead support the hypothesis that these oxons bind to a secondary site on acetylcholinesterase, leading to activation/inhibition of the catalytic site, depending on the nature of the substrate and inhibitor. Additionally, these data raise questions regarding the adequacy of estimating risk of low levels of insecticide exposure from direct extrapolation of insecticide dose-response curves since the capacity of individual oxon molecules at low oxon levels could be greater than individual oxon molecules in vivo associated with the dose-response curve

  20. An evaluation of the inhibition of human butyrylcholinesterase and acetylcholinesterase by the organophosphate chlorpyrifos oxon

    International Nuclear Information System (INIS)

    Shenouda, Josephine; Green, Paula; Sultatos, Lester

    2009-01-01

    Acetylcholinesterase (EC 3.1.1.7) and butyrylcholinesterase (EC 3.1.1.8) are enzymes that belong to the superfamily of α/β-hydrolase fold proteins. While they share many characteristics, they also possess many important differences. For example, whereas they have about 54% amino acid sequence identity, the active site gorge of acetylcholinesterase is considerably smaller than that of butyrylcholinesterase. Moreover, both have been shown to display simple and complex kinetic mechanisms, depending on the particular substrate examined, the substrate concentration, and incubation conditions. In the current study, incubation of butyrylthiocholine in a concentration range of 0.005-3.0 mM, with 317 pM human butyrylcholinesterase in vitro, resulted in rates of production of thiocholine that were accurately described by simple Michaelis-Menten kinetics, with a K m of 0.10 mM. Similarly, the inhibition of butyrylcholinesterase in vitro by the organophosphate chlorpyrifos oxon was described by simple Michaelis-Menten kinetics, with a k i of 3048 nM -1 h -1 , and a K D of 2.02 nM. In contrast to inhibition of butyrylcholinesterase, inhibition of human acetylcholinesterase by chlorpyrifos oxon in vitro followed concentration-dependent inhibition kinetics, with the k i increasing as the inhibitor concentration decreased. Chlorpyrifos oxon concentrations of 10 and 0.3 nM gave k i s of 1.2 and 19.3 nM -1 h -1 , respectively. Although the mechanism of concentration-dependent inhibition kinetics is not known, the much smaller, more restrictive active site gorge of acetylcholinesterase almost certainly plays a role. Similarly, the much larger active site gorge of butyrylcholinesterase likely contributes to its much greater reactivity towards chlorpyrifos oxon, compared to acetylcholinesterase.

  1. Brain acetylcholinesterase activity controls systemic cytokine levels through the cholinergic anti-inflammatory pathway

    Science.gov (United States)

    Pavlov, Valentin A.; Parrish, William R.; Rosas-Ballina, Mauricio; Ochani, Mahendar; Puerta, Margot; Ochani, Kanta; Chavan, Sangeeta; Al-Abed, Yousef; Tracey, Kevin J.

    2015-01-01

    The excessive release of cytokines by the immune system contributes importantly to the pathogenesis of inflammatory diseases. Recent advances in understanding the biology of cytokine toxicity led to the discovery of the “cholinergic anti-inflammatory pathway,” defined as neural signals transmitted via the vagus nerve that inhibit cytokine release through a mechanism that requires the alpha7 subunit-containing nicotinic acetylcholine receptor (α7nAChR). Vagus nerve regulation of peripheral functions is controlled by brain nuclei and neural networks, but despite considerable importance, little is known about the molecular basis for central regulation of the vagus nerve-based cholinergic anti-inflammatory pathway. Here we report that brain acetylcholinesterase activity controls systemic and organ specific TNF production during endotoxemia. Peripheral administration of the acetylcholinesterase inhibitor galantamine significantly reduced serum TNF levels through vagus nerve signaling, and protected against lethality during murine endotoxemia. Administration of a centrally-acting muscarinic receptor antagonist abolished the suppression of TNF by galantamine, indicating that suppressing acetylcholinesterase activity, coupled with central muscarinic receptors, controls peripheral cytokine responses. Administration of galantamine to α7nAChR knockout mice failed to suppress TNF levels, indicating that the α7nAChR-mediated cholinergic anti-inflammatory pathway is required for the anti-inflammatory effect of galantamine. These findings show that inhibition of brain acetylcholinesterase suppresses systemic inflammation through a central muscarinic receptor-mediated and vagal- and α7nAChR-dependent mechanism. Our data also indicate that a clinically used centrally-acting acetylcholinesterase inhibitor can be utilized to suppress abnormal inflammation to therapeutic advantage. PMID:18639629

  2. Chlamydia psittaci and C. avium in feral pigeon (Columba livia domestica) droppings in two cities in the Netherlands

    NARCIS (Netherlands)

    Burt, Sara A.; Röring, Romy E.; Heijne, Marloes

    2018-01-01

    Background: Feral pigeons (Columba livia domestica) live and breed in many city centres and contact with their droppings can be a hazard for human health if the birds carry Chlamydia psittaci. Objective: The aim of this study was to establish whether pigeon droppings in two Dutch cities (Utrecht and

  3. Genetic variability in apple fruit polyphenol composition in Malus × domestica and Malus sieversii germplasm grown in New Zealand.

    Science.gov (United States)

    Volz, Richard K; McGhie, Tony K

    2011-11-09

    Variations in the concentrations of flavan-3-ol, oligomeric procyanidin, chlorogenic acid, dihydrochalcone, flavonol, and anthocyanin polyphenol groups and total polyphenols were examined in the fruit peel and cortical flesh of 93 (80 Malus × domestica and 13 Malus sieversii) apple genotypes in at least 1 year between 2003 and 2005 grown at one site in New Zealand (NZ). Differences among genotypes accounted for 46-97% of the total variation in the concentrations of total polyphenols and each of the individual phenol groups in the flesh and peel in both species, whereas effects of year and genotype × year were minimal, except for peel flavonols in M. × domestica and flesh flavonols in both species. In these cases, differences among genotypes accounted for less than 30% of the total variation, which was less than the variation found for the interaction between genotype and year. Total polyphenol concentrations among genotypes were spread over a 7- and 9-fold range in the flesh and a 4- and 3-fold range in the peel of M. sieversii and M. × domestica, respectively, with the spread in concentrations of individual polyphenol groups in each tissue and within each species varying from a 2-fold to over a 500-fold range. Higher concentrations were generally found in M. sieversii. In M. × domestica, cultivars and breeding selections originating in NZ had lower average flesh and peel total polyphenols and chlorogenic acid than older cultivars previously imported into NZ from overseas countries.

  4. ‘Fuji’ apple (Malus domestica Borkh) volatile production during high pCO2 controlled atmosphere storage

    Science.gov (United States)

    ‘Fuji’apple [Malus sylvestris var. domestica (Borkh.) Mansf.] volatile compound dynamics were characterized during cold storage in air or at low pO2 controlled atmosphere (CA) with up to 5 kPa CO2. Volatile compounds in storage chambers were adsorbed onto solid sorbent traps and analyzed by GC-MS....

  5. 'HoneySweet' (C5), the first genetically engineered Plum pox virus-resistant plum (Prunus domestica L.) cultivar

    Science.gov (United States)

    ‘HoneySweet’ plum was released by the U.S. Department of Agriculture, Agricultural Research Service, to provide U.S. growers and P. domestica plum breeders with a high fruit quality plum cultivar resistant to Plum pox virus (PPV). ‘HoneySweet’ was developed through genetic engineering utilizing the...

  6. The mitochondrial genomes of the barklice, Lepinotus reticulatus and Dorypteryx domestica (Psocodea: Trogiomorpha): Insight into phylogeny of the order Psocodea.

    Science.gov (United States)

    Feng, Shiqian; Stejskal, Václav; Wang, Yannan; Li, Zhihong

    2018-05-05

    The order Psocodea which has incorporated the two former orders Psocoptera (barklice and booklice) and Phthiraptera (parasitic lice) attracts much attention for its unusual mitochondrial (mt) genome rearrangements. Available phylogenetic analysis for Psocodea is subjected to partial taxa and a complete one is needed. To further explore the genome rearrangement and phylogeny in Psocodea, we sequenced the mt genomes of two barklice, Lepinotus reticulatus (collected from China) and Dorypteryx domestica (collected from Czech Republic). Both of newly sequenced barklice had typical one-chromosome mt genomes and the same mt gene arrangement with the reported Lepidopsocidae sp. The mt genomes of L. reticulatus and D. domestica contained 37 genes typical of bilateral animals. In contrast with the recent report mt genome of D. domestica, our strain was found with many single nucleotide polymorphisms in intra-specific difference. Phylogenetic relationships were inferred from all available mt genomes of Psocodea data using Maximum Likelihood and Bayesian methods. The mt genome of L. reticulatus is the first representative with complete sequences of the family Trogiidae and our D. domestica data enriched the family Psyllipsocidae, which will contribute to the further study of mt gene rearrangement and phylogeny of Psocodea. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. Radiation induced cerebellum impairments in Swiss albino mice and its modulation by dietary Prunus domestica

    International Nuclear Information System (INIS)

    Sharma, Garima; Sisodia, Rashmi

    2012-01-01

    To study the biochemical, quantitative histopathological and behavioural changes after 5 Gy whole body irradiation and its modulation by supplementation of Prunus domestica extract (PDE) for 15 consecutive days on male Swiss albino. For this study healthy mice from an inbred colony were divided into five groups: (i) Control; (ii) PDE treated - mice in this group were orally supplemented with PDE (400 mg/kg body weight (bw)/day) once daily for 15 consecutive days; (iii) Irradiated-mice were whole body exposed to 5 Gy irradiated; (iv) PDE + irradiated-mice in this group were orally supplemented PDE for 15 days (once a day) prior to irradiation; and (v) irradiated+PDE -mice in this group were administered PDE orally for 15 days (once a day) consequently after irradiation. Marked radiation induced changes in the amount of cerebellar lipid peroxidation (LPO), glutathione (GSH), protein, superoxide dismutase (SOD), catalase and histopathological changes (molecular layer, granular layer and purkinje cell numbers) could be significantly ameliorated supplementation of PDE prior/post irradiation. Radiation induced deficits in learning and memory were also significantly ameliorated. PDE was found to have strong radical scavenging activity in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and also showed in vitro radioprotective activity. The result of present study showed that prior/post-supplementation of Prunus domestica has radioprotective potential as well as neuroprotective properties against the radiation. (author)

  8. The effect of gamma irradiation on curcumin component of Curcuma domestica

    Science.gov (United States)

    Chosdu, R.; Erizal; Iriawan, T.; Hilmy, N.

    1995-02-01

    The effect of gamma irradiation on curcumin component of Curcuma domestica rhizome were investigated. Pure curcumin, sliced and powdered rhizome with 10% of moisture content were irradiated at 0, 10, 30 and 50 kGy (dose rate of 6 kGy/h). Curcumin content was analysed using HPLC method and ESR spectra. Results show that free radicals are already present in unirradiated rhizome. Gamma irradiation at the doses of 10, 30 and 50 kGy induced the free radicals formation of pure curcumin and Curcuma domestica rhizome. The ESR spectra of irradiated rhizome gave a very similar spectra to the signal of irradiated pure curcumin. The percentage of free radicals intensity from pure curcumin was very stable at room temperature up to 670 hours of storage. However, the percentage intensity of free radicals in the irradiated rhizome were decay during storage. Irradiation treatment and storage time did not give a significant change on curcumin content, water activity, pH and moisture content of rhizome investigated.

  9. The effect of gamma irradiation on curcumin component of Curcuma domestica

    Energy Technology Data Exchange (ETDEWEB)

    Chosdu, R.E.; Erizal; Iriawan, T.; Hilmy, N. [National Atomic Energy Agency, Jakarta (Indonesia). Center for Applications of Isotopes and Radiation

    1995-10-01

    The effect of gamma irradiation on curcumin component of Curcuma domestica rhizome were investigated. Pure curcumin, sliced and powdered rhizome with 10% of moisture content were irradiated at 0, 10, 30 and 50 kGy (dose rate of 6 kGy/h). Curcumin content was analysed using HPLC method and ESR spectra. Results show that free radicals are already present in unirradiated rhizome. Gamma irradiation at the doses of 10, 30 and 50 kGy induced the free radicals formation of pure curcumin and curcuma domestica rhizome. The ESR spectra of irradiated rhizome gave a very similar spectra to the signal of irradiated pure curcumin. The percentage of free radicals intensity from pure curcumin was very stable at room temperature up to 670 hours of storage. However, the percentage intensity of free radicals in the irradiated rhizome were decay during storage. Irradiation treatment and storage time did not give a significant change on curcumin content, water activity, pH and moisture content of rhizome investigated. (Author).

  10. The effect of gamma irradiation on curcumin component of Curcuma domestica

    International Nuclear Information System (INIS)

    Chosdu, R.E.; Erizal; Iriawan, T.; Hilmy, N.

    1995-01-01

    The effect of gamma irradiation on curcumin component of Curcuma domestica rhizome were investigated. Pure curcumin, sliced and powdered rhizome with 10% of moisture content were irradiated at 0, 10, 30 and 50 kGy (dose rate of 6 kGy/h). Curcumin content was analysed using HPLC method and ESR spectra. Results show that free radicals are already present in unirradiated rhizome. Gamma irradiation at the doses of 10, 30 and 50 kGy induced the free radicals formation of pure curcumin and curcuma domestica rhizome. The ESR spectra of irradiated rhizome gave a very similar spectra to the signal of irradiated pure curcumin. The percentage of free radicals intensity from pure curcumin was very stable at room temperature up to 670 hours of storage. However, the percentage intensity of free radicals in the irradiated rhizome were decay during storage. Irradiation treatment and storage time did not give a significant change on curcumin content, water activity, pH and moisture content of rhizome investigated. (Author)

  11. Characterisation of Sorbus domestica L. Bark, Fruits and Seeds: Nutrient Composition and Antioxidant Activity

    Directory of Open Access Journals (Sweden)

    Boris Majić

    2015-01-01

    Full Text Available The aim of this work is to assess the nutritional value of service tree (Sorbus domestica L. bark, fruit exocarp and mesocarp, and seeds by establishing the levels of macro- and microelements, total phenolics, flavonoids and tannins. Our results revealed that all of the tested service tree samples were rich in potassium. Bark was the best source of calcium and zinc, while seeds were the best source of magnesium. Compared to the bark and seeds, fruit exocarp and mesocarp contained significantly lower amounts of these three elements. Immature exocarp and bark contained the highest amounts of total phenolics and showed the highest antioxidant activity. Maturation significantly decreased the amount of total phenolics in fruits, as well as the antioxidant activity of total phenolics and total tannins from exocarp, but not from mesocarp. Exocarp was the richest in total flavonoids. Based on the obtained data, we have concluded that the under-utilised species S. domestica L. could serve as an important source of mineral elements and antioxidants in the human diet.

  12. Chlorophyll Catabolites in Senescent Leaves of the Plum Tree (Prunus domestica).

    Science.gov (United States)

    Erhart, Theresia; Mittelberger, Cecilia; Vergeiner, Clemens; Scherzer, Gerhard; Holzner, Barbara; Robatscher, Peter; Oberhuber, Michael; Kräutler, Bernhard

    2016-11-01

    In cold extracts of senescent leaves of the plum tree (Prunus domestica ssp. domestica), six colorless non-fluorescent chlorophyll catabolites (NCCs) were characterized, named Pd-NCCs. In addition, several minor NCC fractions were tentatively classified. The structure of the most polar one of the NCCs, named Pd-NCC-32, featured an unprecedented twofold glycosidation pattern. Three of the NCCs are also functionalized at their 3 2 -position by a glucopyranosyl group. In addition, two of these glycosidated NCCs carry a dihydroxyethyl group at their 18-position. In the polar Pd-NCC-32, the latter group is further glycosidated at the terminal 18 2 -position. Four other major Pd-NCCs and one minor Pd-NCC were identified with five NCCs from higher plants known to belong to the 'epi'-series. In addition, tentative structures were derived for two minor fractions, classified as yellow chlorophyll catabolites, which represented (formal) oxidation products of two of the observed Pd-NCCs. The chlorophyll catabolites in leaves of plum feature the same basic structural pattern as those found in leaves of apple and pear trees. © 2016 The Authors. Chemistry & Biodiversity Published by Wiley-VHCA AG, Zurich, Switzerland.

  13. Boron toxicity causes multiple effects on Malus domestica pollen tube growth

    Directory of Open Access Journals (Sweden)

    Kefeng eFang

    2016-02-01

    Full Text Available Boron is an essential micronutrient for plants. However, boron is also toxic to cells at high concentrations, although the mechanism of this stress is not known. This study aimed to evaluate the effect of boron stress on Malus domestica pollen tube growth and its possible regulatory pathway. Our results show that a high concentration of boron inhibited pollen germination and tube growth and led to the morphological abnormality of pollen tubes. Fluorescent labeling coupled with a scanning ion-selective electrode technique detected that boron stress could decrease [Ca2+]c and induce the disappearance of the [Ca2+]c gradient, which are critical for pollen tube polar growth. Actin filaments were therefore altered by boron stress. Immuno-localization and fluorescence labeling, together with Fourier-transform infrared analysis (FTIR, suggested that boron stress influenced the accumulation and distribution of callose, de-esterified pectins, esterified pectins and arabinogalactan proteins in pollen tubes. All of the above results provide new insights into the regulatory role of boron in pollen tube development. In summary, boron likely plays a structural and regulatory role in relation to [Ca2+]c, actin cytoskeleton and cell wall components and thus regulates Malus domestica pollen germination and tube polar growth.

  14. Boron Toxicity Causes Multiple Effects on Malus domestica Pollen Tube Growth.

    Science.gov (United States)

    Fang, Kefeng; Zhang, Weiwei; Xing, Yu; Zhang, Qing; Yang, Liu; Cao, Qingqin; Qin, Ling

    2016-01-01

    Boron is an important micronutrient for plants. However, boron is also toxic to cells at high concentrations, although the mechanism of this toxicity is not known. This study aimed to evaluate the effect of boron toxicity on Malus domestica pollen tube growth and its possible regulatory pathway. Our results showed that a high concentration of boron inhibited pollen germination and tube growth and led to the morphological abnormality of pollen tubes. Fluorescent labeling coupled with a scanning ion-selective electrode technique detected that boron toxicity could decrease [Ca(2+)]c and induce the disappearance of the [Ca(2+)]c gradient, which are critical for pollen tube polar growth. Actin filaments were therefore altered by boron toxicity. Immuno-localization and fluorescence labeling, together with fourier-transform infrared analysis, suggested that boron toxicity influenced the accumulation and distribution of callose, de-esterified pectins, esterified pectins, and arabinogalactan proteins in pollen tubes. All of the above results provide new insights into the regulatory role of boron in pollen tube development. In summary, boron likely plays a structural and regulatory role in relation to [Ca(2+)]c, actin cytoskeleton and cell wall components and thus regulates Malus domestica pollen germination and tube polar growth.

  15. Antioxidant, antimicrobial activity and mineral composition of low-temperature fractioning products of Malus domestica Borkh (common Antonovka

    Directory of Open Access Journals (Sweden)

    Elena Kuznetsova

    2017-01-01

    Full Text Available The low-temperature fractionation of fruit Malus domestica Borkh (Common Antonovka has been performed. We obtained by fractionation the biologically active products that are the dehydrated concentrate of juice and the powder of pomace fibers. Use of low temperature minimizes biological value losses during processing. These fractions of fruit Malus domestica Borkh (Common Antonovka are experimentally studied. It is found that the fractions have high antioxidant activity and include bioflavonoids and organic and phenol carboxylic acids. Analysis of chromatograms showed availability of the identical compounds in the products of low-temperature fractionation. Sodium and potassium are part of the cells of biological systems as highly mobile ionic forms. Therefore, these elements prevail in the concentrated juice. Iron, manganese, copper, and zinc are biogenic trace elements or components of enzyme systems and are evenly distributed as in plant cell walls as well in protoplasm. It follows from the results of the study of the mineral composition that the products of the low-temperature fractioning can be used for a functional food as a result of its high content of magnesium and iron. The low-temperature fractionation of fruit Malus domestica Borkh (Common Antonovka has antimicrobial activity against the standard strains of spoilage: Bacillus subtilis VKM-B-501, Micrococcus luteus VKM-As-2230, Aspergillus flavus VKM-F-1024, Penicillium expansion VKM-F-275, Mucor mucedo VKM- F-1257, Rhizopus stolonifer VKM- F-2005. Experimental data show that the products of low-temperature fractioning of Malus domestica Borkh (Common Antonovka inhibit microorganism's growth. The detected composition of Malus domestica Borkh (Common Antonovka fractions allows using these products as natural additives in food technology to maintain and increase period of storage and also for preventive nutrition.

  16. Anti-acetylcholinesterase and Antioxidant Activity of Essential Oils from Hedychium gardnerianum Sheppard ex Ker-Gawl

    OpenAIRE

    Arruda, Miguel; Viana, Hugo; Rainha, Nuno; Neng, Nuno R.; Rosa, José Silvino; Nogueira, José M. F.; Barreto, Maria do Carmo

    2012-01-01

    Acetylcholinesterase inhibition, antioxidant and cytotoxic activities of Hedychium gardnerianum leaf essential oils from S. Miguel Island were determined. All the oils inhibited acetylcholinesterase, with IC50 values of approximately 1 mg/mL, showing no statistical differences between collection sites. Three oils presented mixed inhibition, whilst one was almost truly competitive. This activity can be attributed to the presence of sesquiterpenes, which constituted more than 60% of the composi...

  17. Inhibition of Acetylcholinesterase and Fatty Acid Composition in Theobroma grandiflorum Seeds

    Directory of Open Access Journals (Sweden)

    Casandra Valentina Itriago

    2017-07-01

    Full Text Available Theobroma grandiflorum is an important fruit tree from Sterculiaceae family, native to the Brazilian Amazon, known in the region as cupuaçu. The seeds have a high fat content (24% with characteristics that resemble those of cocoa (Theobroma cacao butter with potential applications in the cosmetic, pharmaceutical and food industries. The main objective of this work was to explore the seed fats from T. grandiflorum that were analyzed for fatty acid composition by Gas Chromatography with Flame Ionization Detector (GC-FID and to analyze their activity for acetylcholinesterase inhibition. Chromatographic analysis provided detection of nine fatty acids. The major fatty acids found in the species were oleic (40.0%, stearic (32.7%, arachidic (10.4% and palmitic (8.0%. The acetylcholinesterase inhibition by fats from seeds was over 40.48%. DOI: http://dx.doi.org/10.17807/orbital.v0i0.894 

  18. Alpha-Bungarotoxin labeling and acetylcholinesterase localization at the Mauthner fiber giant synapse in the hatchetfish

    International Nuclear Information System (INIS)

    Day, J.W.; Hall, D.H.; Hall, L.M.; Bennett, M.V.

    1983-01-01

    Autoradiographic and histochemical techniques have been used to characterize further the pharmacology of transmission at the Mauthner fiber giant synapse of the South American hatchetfish. [ 125 I]alpha-Bungarotoxin was applied to hatchetfish medullae and a standard autoradiographic procedure was carried out on 3- to 4-microns sections of glutaraldehyde-fixed tissue. All Mauthner fiber giant synapses, as identified by light microscopic criteria, had closely associated silver grains. Labeling was blocked by d-tubocurarine. Glutaraldehyde-fixed slices of hatchetfish medulla were stained histochemically for acetylcholinesterase; all giant synapses that could be identified in the light microscope showed heavy deposits of reaction product. Staining was blocked by diisopropyl-fluorophosphate, which inhibits both pseudocholinesterase and acetylcholinesterase, but was not blocked by tetraisopropylpyrophosphoramide, a specific pseudocholinesterase inhibitor. This evidence strongly supports the suggestion that the Mauthner fiber giant synapse is nicotinic cholinergic

  19. alpha-Bungarotoxin labeling and acetylcholinesterase localization at the Mauthner fiber giant synapse in the hatchetfish

    International Nuclear Information System (INIS)

    Day, J.W.; Hall, D.H.; Hall, L.M.; Bennett, M.V.

    1983-01-01

    Autoradiographic and histochemical techniques have been used to characterize further the pharmacology of transmission at the Mauthner fiber giant synapse of the South American hatchetfish. [ 125 I]alpha-Bungarotoxin was applied to hatchetfish medullae and a standard autoradiographic procedure was carried out on 3- to 4-microns sections of glutaraldehyde-fixed tissue. All Mauthner fiber giant synapses, as identified by light microscopic criteria, had closely associated silver grains. Labeling was blocked by d-tubocurarine. Glutaraldehyde-fixed slices of hatchetfish medulla were stained histochemically for acetylcholinesterase; all giant synapses that could be identified in the light microscope showed heavy deposits of reaction product. Staining was blocked by diisopropyl-fluorophosphate, which inhibits both pseudocholinesterase and acetylcholinesterase, but was not blocked by tetraisopropylpyrophosphoramide, a specific pseudocholinesterase inhibitor. This evidence strongly supports the suggestion that the Mauthner fiber giant synapse is nicotinic cholinergic

  20. A New Acetylcholinesterase Inhibitor from Green Glycosylation of Trachyloban-19-oic Acid by Mucor plumbeus.

    Science.gov (United States)

    Santos, Gabriel F Dos; Takahashi, Jacqueline A

    2017-01-01

    The in vitro metabolism of a widespread natural product, trachyloban-19-oic acid (1), by the fungal species Mucor plumbeus was studied in a sucrose-yeast liquid medium. Two products were isolated, and their structures were determined by spectroscopic means as 7β-hydroxytrachyloban-19-oic acid (5) and trachyloban-19-O-β-D-glucopyranosyl ester (6). To the best of our knowledge, compound 6 is herein reported by the first time in the literature. These compounds were assayed for acetylcholinesterase inhibition along with some related compounds. Compound 6 showed the highest acetylcholinesterase inhibitory activity at 10000 µg/mL among the tested compounds, a result (92.89%) comparable to the activity of the positive control, galanthamine (94.21%). Therefore, biotransformation of the natural product 1 by M. plumbeus produced a novel compound with potential as a new lead to develop anti-Alzheimer medicines.

  1. A New Acetylcholinesterase Inhibitor from Green Glycosylation of Trachyloban-19-oic Acid by Mucor plumbeus

    Directory of Open Access Journals (Sweden)

    GABRIEL F. DOS SANTOS

    2017-10-01

    Full Text Available ABSTRACT The in vitro metabolism of a widespread natural product, trachyloban-19-oic acid (1, by the fungal species Mucor plumbeus was studied in a sucrose-yeast liquid medium. Two products were isolated, and their structures were determined by spectroscopic means as 7β-hydroxytrachyloban-19-oic acid (5 and trachyloban-19-O-β-D-glucopyranosyl ester (6. To the best of our knowledge, compound 6 is herein reported by the first time in the literature. These compounds were assayed for acetylcholinesterase inhibition along with some related compounds. Compound 6 showed the highest acetylcholinesterase inhibitory activity at 10000 µg/mL among the tested compounds, a result (92.89% comparable to the activity of the positive control, galanthamine (94.21%. Therefore, biotransformation of the natural product 1 by M. plumbeus produced a novel compound with potential as a new lead to develop anti-Alzheimer medicines.

  2. Alpha-Bungarotoxin labeling and acetylcholinesterase localization at the Mauthner fiber giant synapse in the hatchetfish

    Energy Technology Data Exchange (ETDEWEB)

    Day, J.W.; Hall, D.H.; Hall, L.M.; Bennett, M.V.

    1983-02-01

    Autoradiographic and histochemical techniques have been used to characterize further the pharmacology of transmission at the Mauthner fiber giant synapse of the South American hatchetfish. (/sup 125/I)alpha-Bungarotoxin was applied to hatchetfish medullae and a standard autoradiographic procedure was carried out on 3- to 4-microns sections of glutaraldehyde-fixed tissue. All Mauthner fiber giant synapses, as identified by light microscopic criteria, had closely associated silver grains. Labeling was blocked by d-tubocurarine. Glutaraldehyde-fixed slices of hatchetfish medulla were stained histochemically for acetylcholinesterase; all giant synapses that could be identified in the light microscope showed heavy deposits of reaction product. Staining was blocked by diisopropyl-fluorophosphate, which inhibits both pseudocholinesterase and acetylcholinesterase, but was not blocked by tetraisopropylpyrophosphoramide, a specific pseudocholinesterase inhibitor. This evidence strongly supports the suggestion that the Mauthner fiber giant synapse is nicotinic cholinergic.

  3. alpha-Bungarotoxin labeling and acetylcholinesterase localization at the Mauthner fiber giant synapse in the hatchetfish

    Energy Technology Data Exchange (ETDEWEB)

    Day, J.W.; Hall, D.H.; Hall, L.M.; Bennett, M.V.

    1983-02-01

    Autoradiographic and histochemical techniques have been used to characterize further the pharmacology of transmission at the Mauthner fiber giant synapse of the South American hatchetfish. (/sup 125/I)alpha-Bungarotoxin was applied to hatchetfish medullae and a standard autoradiographic procedure was carried out on 3- to 4-microns sections of glutaraldehyde-fixed tissue. All Mauthner fiber giant synapses, as identified by light microscopic criteria, had closely associated silver grains. Labeling was blocked by d-tubocurarine. Glutaraldehyde-fixed slices of hatchetfish medulla were stained histochemically for acetylcholinesterase; all giant synapses that could be identified in the light microscope showed heavy deposits of reaction product. Staining was blocked by diisopropyl-fluorophosphate, which inhibits both pseudocholinesterase and acetylcholinesterase, but was not blocked by tetraisopropylpyrophosphoramide, a specific pseudocholinesterase inhibitor. This evidence strongly supports the suggestion that the Mauthner fiber giant synapse is nicotinic cholinergic.

  4. Chemical composition and acetylcholinesterase inhibitory activity of Artemisia maderaspatana essential oil.

    Science.gov (United States)

    Jyotshna; Srivastava, Nidhi; Singh, Bhuwanendra; Chanda, Debabrata; Shanker, Karuna

    2015-01-01

    To date, there are no reports to validate the Indian traditional and folklore claims of Artemisia maderaspatana L. (syn. Grangea maderaspatana L.) (Asteraceae) for the treatment of Alzheimer's disease. The present study characterizes the volatile components (non-polar compounds) of A. maderaspatana and evaluates its acetylcholinesterase inhibition potential. The essential oils (yield 0.06% v/w) were obtained from fresh aerial part of A. maderaspatana. The characterization of volatile components (non-polar compounds) was performed by GC-MS data and with those of reference compounds compiled in the spectral library of in-house database. The in vitro acetylcholinesterase (AChE) inhibition of the volatile organic constituents (VOC's) of A. maderaspatana aerial part was evaluated in varying concentration ranges (0.70-44.75 µg/mL) with Ellman's method. The major components were α-humulene (46.3%), β-caryophyllene (9.3%), α-copaene (8.2%), β-myrcene (4.3%), Z(E)-α-farnesene (3.7%), and calarene (3.5%). Chemical variability among other Artemisia spp. from different climatic regions of India and countries namely Iran and France was observed. The experimental results showed that diverse volatile organic constituents of A. maderaspatana have significant acetylcholinesterase inhibitory activity (an IC50 value of 31.33 ± 1.03 µg/mL). This is the first report on the inhibition of acetylcholinesterase properties of essential oil of A. maderaspatana obtained from fresh aerial part. The present results indicate that essential oil of A. maderaspatana isolated from the northern region of India could inhibit AChE moderately. Therefore, the possibility of novel AChE inhibitors might exist in VOCs of this plant.

  5. Acute administration of fenproporex increased acetylcholinesterase activity in brain of young rats

    OpenAIRE

    TEODORAK, BRENA P.; FERREIRA, GABRIELA K.; SCAINI, GISELLI; WESSLER, LETÍCIA B.; HEYLMANN, ALEXANDRA S.; DEROZA, PEDRO; VALVASSORI, SAMIRA S.; ZUGNO, ALEXANDRA I.; QUEVEDO, JOÃO; STRECK, EMILIO L.

    2015-01-01

    Fenproporex is the second most commonly amphetamine-based anorectic consumed worldwide; this drug is rapidly converted into amphetamine, in vivo, and acts by increasing dopamine levels in the synaptic cleft. Considering that fenproporex effects on the central nervous system are still poorly known and that acetylcholinesterase is a regulatory enzyme which is involved in cholinergic synapses and may indirectly modulate the release of dopamine, the present study investigated the effects of acute...

  6. Methyl jasmonate enhances memory performance through inhibition of oxidative stress and acetylcholinesterase activity in mice.

    Science.gov (United States)

    Eduviere, Anthony T; Umukoro, S; Aderibigbe, Adegbuyi O; Ajayi, Abayomi M; Adewole, Folashade A

    2015-07-01

    Current research effort focuses on the development of safer natural compounds with multipronged mechanisms of action that could be used to ameliorate memory deficits in patients with Alzheimer's disease, as cure for the disease still remains elusive. In this study, we evaluated the effect of methyl jasmonate (MJ), a naturally occurring bioactive compound on memory, acetylcholinesterase activity and biomarkers of oxidative stress in mice. Male Swiss mice were treated with intraperitoneal injection of MJ (10-40 mg/kg) alone or in combination with scopolamine (3mg/kg) once daily for 7 days. Thirty minutes after the last treatment, memory functions were assessed using Y-maze and object recognition tests. Thereafter, acetylcholinesterase activity and levels of biomarkers of oxidative stress were assessed in mice brains using standard biochemical procedures. MJ significantly enhanced memory performance and reversed scopolamine-induced cognitive impairment in mice. MJ demonstrated significant inhibition of acetylcholinesterase activity suggesting increased cholinergic neurotransmission. It further decreased malondialdehyde concentrations in mouse brain indicating antioxidant activity. Moreover, MJ significantly increased glutathione levels and activity of antioxidant enzymes (catalase and superoxide dismutase) in mice brains. The increased oxidative stress; evidenced by elevated levels of malondialdehyde and decreased antioxidant defense systems in scopolamine-treated mice was attenuated by MJ. The results of this study suggest that MJ may be useful in conditions associated with memory dysfunctions or age-related cognitive decline. The positive effect of MJ on memory may be related to inhibition of oxidative stress and enhancement of cholinergic neurotransmission through inhibition of acetylcholinesterase activity. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Phytochemical Characterization, Antibacterial, Acetylcholinesterase Inhibitory and Cytotoxic Properties of Cryptostephanus vansonii, an Endemic Amaryllid

    Czech Academy of Sciences Publication Activity Database

    Moyo, M.; Aremu, A.O.; Chukwujekwu, J. C.; Grúz, Jiří; Skořepa, Jiří; Doležal, Karel; Katsvanga, C. A. T.; Van Staden, J.

    2017-01-01

    Roč. 31, č. 5 (2017), s. 713-720 ISSN 0951-418X R&D Projects: GA MŠk(CZ) LO1204; GA MŠk LK21306 Institutional support: RVO:61389030 Keywords : in-vitro * alzheimers-disease * alkaloids * extracts * antioxidant * coumarins * apoptosis * phylogeny * medicine * bacteria * acetylcholinesterase inhibition * Amaryllidaceae * antibacterial * cytotoxicity * flavonoids * phenolic acids Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Plant sciences, botany Impact factor: 3.092, year: 2016

  8. Antioxidant activity, acetylcholinesterase and tyrosinase inhibitory potential of Pulmonaria officinalis and Centarium umbellatum extracts

    Directory of Open Access Journals (Sweden)

    Elena Neagu

    2018-03-01

    Full Text Available In this study several investigations and tests were performed to determine the antioxidant activity and the acetylcholinesterase and tyrosinase inhibitory potential of Pulmonaria officinalis and Centarium umbellatum aqueous extracts (10% mass and ethanolic extracts (10% mass and 70% ethanol, respectively. Moreover, for each type of the prepared extracts of P. officinalis and of C. umbellatum the content in the biologically active compounds – polyphenols, flavones and proanthocyanidins was determined. The antioxidant activity was assessed using two methods, namely the 2,2-diphenyl-1-picrylhydrazyl (DPPH assay and reducing power assay. The analyzed plant extracts showed a high acetylcholinesterase and tyrosinase inhibitory activity in the range of 72.24–94.24% (at the highest used dose – 3 mg/mL, 66.96% and 94.03% (at 3 mg/mL, respectively correlated with a high DPPH radical inhibition – 70.29–84.9% (at 3 mg/mL. These medicinal plants could provide a potential natural source of bioactive compounds and could be beneficial to the human health, especially in the neurodegenerative disorders and as sources of natural antioxidants in food industry. Keywords: Acetylcholinesterase inhibitory activity, Tyrosinase inhibitory activity, Antioxidant activity, Pulmonaria officinalis and Centarium umbellatum

  9. Acetylcholinesterase inhibitors rapidly activate Trk neurotrophin receptors in the mouse hippocampus

    Science.gov (United States)

    Autio, Henri; Mätlik, Kert; Rantamäki, Tomi; Lindemann, Lothar; Hoener, Marius C; Chao, Moses; Arumäe, Urmas; Castrén, Eero

    2014-01-01

    Acetylcholinesterase inhibitors are first-line therapies for Alzheimer's disease. These drugs increase cholinergic tone in the target areas of the cholinergic neurons of the basal forebrain. Basal forebrain cholinergic neurons are dependent upon trophic support by nerve growth factor (NGF) through its neurotrophin receptor, TrkA. In the present study, we investigated whether the acetylcholinesterase inhibitors donepezil and galantamine could influence neurotrophin receptor signaling in the brain. Acute administration of donepezil (3 mg/kg, i.p.) led to the rapid autophosphorylation of TrkA and TrkB neurotrophin receptors in the adult mouse hippocampus. Similarly, galantamine dose-dependently (3, 9 mg/kg, i.p.) increased TrkA and TrkB phosphorylation in the mouse hippocampus. Both treatments also increased the phosphorylation of transcription factor CREB and tended to increase the phosphorylation of AKT kinase but did not alter the activity of MAPK42/44. Chronic treatment with galantamine (3 mg/kg, i.p., 14 days), did not induce changes in hippocampal NGF and BDNF synthesis or protein levels. Our findings show that acetylcholinesterase inhibitors are capable of rapidly activating hippocampal neurotrophin signaling and thus suggest that therapies targeting Trk signaling may already be in clinical use in the treatment of AD. PMID:21820453

  10. Upregulation of α7 Nicotinic Receptors by Acetylcholinesterase C-Terminal Peptides

    Science.gov (United States)

    Bond, Cherie E.; Zimmermann, Martina; Greenfield, Susan A.

    2009-01-01

    Background The alpha-7 nicotinic acetylcholine receptor (α7-nAChR) is well known as a potent calcium ionophore that, in the brain, has been implicated in excitotoxicity and hence in the underlying mechanisms of neurodegenerative disorders such as Alzheimer's disease. Previous research implied that the activity of this receptor may be modified by exposure to a peptide fragment derived from the C-terminal region of the enzyme acetylcholinesterase. This investigation was undertaken to determine if the functional changes observed could be attributed to peptide binding interaction with the α7-nAChR, or peptide modulation of receptor expression. Methodology/Principal Findings This study provides evidence that two peptides derived from the C-terminus of acetylcholinesterase, not only selectively displace specific bungarotoxin binding at the α7-nAChR, but also alter receptor binding properties for its familiar ligands, including the alternative endogenous agonist choline. Of more long-term significance, these peptides also induce upregulation of α7-nAChR mRNA and protein expression, as well as enhancing receptor trafficking to the plasma membrane. Conclusions/Significance The results reported here demonstrate a hitherto unknown relationship between the α7-nAChR and the non-enzymatic functions of acetylcholinesterase, mediated independently by its C-terminal domain. Such an interaction may prove valuable as a pharmacological tool, prompting new approaches for understanding, and combating, the process of neurodegeneration. PMID:19287501

  11. Optimization of sol-gel medium for entrapment of acetylcholinesterase enzyme in biosensor for pesticide detection

    Science.gov (United States)

    Wijayanti, S. D.; Rahayu, F. S.; Widyaningsih, T. D.

    2018-03-01

    Pesticides are chemical substances used to kill and control pests or diseases that can damage crops. The use of pesticides should be done precisely because the accumulation of chemicals contained in pesticides can cause various health effects. Therefore, detection of pesticide residues on plants is important to reduce the risk of poisoning due to pesticide residues. Some of the conventional methods that have been done to detect pesticide residues have weaknesses among expensive tools, takes a long time, and are generally performed by trained laboratory technicians. Biosensors are analytical devices that can measure the quantitative or semi-quantitative targets of analyte by utilizing a bioreceptor such as enzyme. Several studies have shown that enzyme-based acetylcholinesterase-based biosensors can be used to detect pesticide residues in vegetable samples. The objective of this research was to get a proper silica based sol-gel formulation with molar ratio of H2O:TEOS and NaOH concentration as immobilization medium of acetylcholinesterase enzyme for biosensor application. Response Surface Methodology (RSM) was used in order to determine the interaction between the parameters studied and resulting responses which were amount and activity of acetylcholinesterase enzyme. Based on the research, the best result for immobilized enzyme activity was shown by molar ratio (H2O: TEOS) 1: 8 and 4 mM NaOH treatment.

  12. Computational study of substrate isotope effect probes of transition state structure for acetylcholinesterase catalysis

    International Nuclear Information System (INIS)

    Sikorski, R.S.; Malany, S.; Seravalli, J.; Quinn, D.M.

    2002-01-01

    Secondary isotope effects for carbonyl addition reactions of methyl thioacetate, acetone and acetaldehyde have been calculated by ab initio quantum mechanical methods in an effect to interpret measured β-deuterium isotope effects on acetylcholinesterase-catalysed hydrolysis of acetylthiocholine. The calculated β-deuterium isotope effect for equilibrium addition of methanol to methyl thioacetate is D3 K eq = 0.965, and the corresponding effect for addition of methoxide ion to methyl thioacetate wherein three waters are hydrogen bonded to the carbonyl oxyanion is D3 K eq = 1.086. Neither of these calculated isotope effects is an inverse as the experimental β-deuterium isotope effect for acetylcholinesterase-catalysed hydrolysis of acetylthiocholine, D3 K eq = 0.90±0.03. Structural comparisons show that the water-solvated methoxide adduct of methyl thioacetate is more expanded than is the natural methanol addition adduct, and suggest that the degree of which the isotope effect is inverse (i.e. less than) is inversely correlated to the degree of expansion of the adduct. A similar correlation of α-deuterium and β-deuterium secondary isotope effects with the degree of expansion of the adducts is found for equilibrium additions of methanol and methoxide ion to acetylaldehyde. These computational results suggest that the markedly inverse β-deuterium isotope effect for the acetylcholinesterase reaction arises from enzymatic compression of the transition state. (author)

  13. Influence of acetylcholinesterase immobilization on the photoluminescence properties of mesoporous silicon surface

    Energy Technology Data Exchange (ETDEWEB)

    Saleem, Muhammad [Department of Chemistry, Kongju National University, Gongju, Chungnam 314-701 (Korea, Republic of); Rafiq, Muhammad; Seo, Sung-Yum [Department of Biology, Kongju National University, Gongju, Chungnam 314-701 (Korea, Republic of); Lee, Ki Hwan, E-mail: khlee@kongju.ac.kr [Department of Chemistry, Kongju National University, Gongju, Chungnam 314-701 (Korea, Republic of)

    2014-07-01

    Acetylcholinesterase immobilized p-type porous silicon surface was prepared by covalent attachment. The immobilization procedure was based on support surface chemical oxidation, silanization, surface activation with cyanuric chloride and finally covalent attachment of free enzyme on the cyanuric chloride activated porous silicon surface. Different pore diameter of porous silicon samples were prepared by electrochemical etching in HF based electrolyte solution and appropriate sample was selected suitable for enzyme immobilization with maximum trapping ability. The surface modification was studied through field emission scanning electron microscope, EDS, FT-IR analysis, and photoluminescence measurement by utilizing the fluctuation in the photoluminescence of virgin and enzyme immobilized porous silicon surface. Porous silicon showed strong photoluminescence with maximum emission at 643 nm and immobilization of acetylcholinesterase on porous silicon surface cause considerable increment on the photoluminescence of porous silicon material while acetylcholinesterase free counterpart did not exhibit any fluorescence in the range of 635–670 nm. The activities of the free and immobilized enzymes were evaluated by spectrophotometric method by using neostigmine methylsulfate as standard enzyme inhibitor. The immobilized enzyme exhibited considerable response toward neostigmine methylsulfate in a dose dependent manner comparable with that of its free counterpart alongside enhanced stability, easy separation from the reaction media and significant saving of enzyme. It was believed that immobilized enzyme can be exploited in organic and biomolecule synthesis possessing technical and economical prestige over free enzyme and prominence of easy separation from the reaction mixture.

  14. Maytenus distichophylla and Salacia crassifolia: source of products with potential acetylcholinesterase inhibition

    Directory of Open Access Journals (Sweden)

    Fernanda L. Ferreira

    Full Text Available ABSTRACT The phytochemical study of the extract leaves from Maytenus distichophylla Mart. and Salacia crassifolia (Mart. ex Schult. G. Don, Celastraceae, resulted in the isolation of 3-oxofriedelane, 3β-hydroxyfriedelane, 3β,24-dihydroxyfriedelane, 3-oxo-28,29-dihydroxyfriedelane, two mixtures of pentacyclic triterpenes (α-amyrin with β-amyrin and 3β-stearyloxy-urs-12-ene with 3β-stearyloxy-olean-12-ene, 3β-palmityloxy-urs-12-ene, the steroid β-sitosterol and its glycosylated derivative β-glucosyl-β-sitosterol, tritriacontanoic acid and the natural polymer gutta percha. The chemical structures of these constituents were established by IR, 1H and 13C NMR spectral data. Crude extracts, the mixtures of triterpenes and the isolated constituents were subjected to in vitro acetylcholinesterase inhibitory evaluation. Acetylcholinesterase inhibitory effect was observed for crude chloroform extract leaves from M. distichophylla (100% and S. crassifolia (97.93 ± 5.63% and for the triterpenes 3β,24-dihydroxyfriedelane (99.05 ± 1.12%, 3-oxo-28,29-dihydroxyfriedelane (90.59 ± 3.76% and 3β-palmityloxy-urs-12-ene (97.93 ± 1.47%. The percent inhibitions induced by these natural products were very similar to those produced by physostigmine (93.94 ± 2.10% a standard acetylcholinesterase inhibitor. Therefore, these results open perspectives for the use of these species as source of compounds with similar physostigmine pharmacological effect.

  15. Hairy-root organ cultures for the production of human acetylcholinesterase

    Directory of Open Access Journals (Sweden)

    Mor Tsafrir S

    2008-12-01

    Full Text Available Abstract Background Human cholinesterases can be used as a bioscavenger of organophosphate toxins used as pesticides and chemical warfare nerve agents. The practicality of this approach depends on the availability of the human enzymes, but because of inherent supply and regulatory constraints, a suitable production system is yet to be identified. Results As a promising alternative, we report the creation of "hairy root" organ cultures derived via Agrobacterium rhizogenes-mediated transformation from human acetylcholinesterase-expressing transgenic Nicotiana benthamiana plants. Acetylcholinesterase-expressing hairy root cultures had a slower growth rate, reached to the stationary phase faster and grew to lower maximal densities as compared to wild type control cultures. Acetylcholinesterase accumulated to levels of up to 3.3% of total soluble protein, ~3 fold higher than the expression level observed in the parental plant. The enzyme was purified to electrophoretic homogeneity. Enzymatic properties were nearly identical to those of the transgenic plant-derived enzyme as well as to those of mammalian cell culture derived enzyme. Pharmacokinetic properties of the hairy-root culture derived enzyme demonstrated a biphasic clearing profile. We demonstrate that master banking of plant material is possible by storage at 4°C for up to 5 months. Conclusion Our results support the feasibility of using plant organ cultures as a successful alternative to traditional transgenic plant and mammalian cell culture technologies.

  16. Influence of acetylcholinesterase immobilization on the photoluminescence properties of mesoporous silicon surface

    International Nuclear Information System (INIS)

    Saleem, Muhammad; Rafiq, Muhammad; Seo, Sung-Yum; Lee, Ki Hwan

    2014-01-01

    Acetylcholinesterase immobilized p-type porous silicon surface was prepared by covalent attachment. The immobilization procedure was based on support surface chemical oxidation, silanization, surface activation with cyanuric chloride and finally covalent attachment of free enzyme on the cyanuric chloride activated porous silicon surface. Different pore diameter of porous silicon samples were prepared by electrochemical etching in HF based electrolyte solution and appropriate sample was selected suitable for enzyme immobilization with maximum trapping ability. The surface modification was studied through field emission scanning electron microscope, EDS, FT-IR analysis, and photoluminescence measurement by utilizing the fluctuation in the photoluminescence of virgin and enzyme immobilized porous silicon surface. Porous silicon showed strong photoluminescence with maximum emission at 643 nm and immobilization of acetylcholinesterase on porous silicon surface cause considerable increment on the photoluminescence of porous silicon material while acetylcholinesterase free counterpart did not exhibit any fluorescence in the range of 635–670 nm. The activities of the free and immobilized enzymes were evaluated by spectrophotometric method by using neostigmine methylsulfate as standard enzyme inhibitor. The immobilized enzyme exhibited considerable response toward neostigmine methylsulfate in a dose dependent manner comparable with that of its free counterpart alongside enhanced stability, easy separation from the reaction media and significant saving of enzyme. It was believed that immobilized enzyme can be exploited in organic and biomolecule synthesis possessing technical and economical prestige over free enzyme and prominence of easy separation from the reaction mixture.

  17. Helminth-bacteria interaction in the gut of domestic pigeon Columba livia domestica.

    Science.gov (United States)

    Biswal, Debraj; Nandi, Anadi Prasad; Chatterjee, Soumendranath

    2016-03-01

    The present paper is an attempt to study the interaction between the helminth parasite and bacteria residing in the gut of domestic pigeon, Columba livia domestica. Biochemical and molecular characterization of the gut bacterial isolate were done and the isolate was identified as Staphylococcus sp. DB1 (JX442510). The interaction of Staphylococcus sp. with Cotugnia cuneata, an intestinal helminth parasite of domestic pigeon was studied on the basis of the difference between 'mean worm burden' of antibiotic treated infected pigeons and infected pigeons without any antibiotic treatment. The ANOVA and Tukey tests of the data obtained showed that antibiotic treatment reduced the mean worm burden significantly. The biochemical properties of Staphylococcus sp. DB1 (JX442510) also showed a mutualistic relationship with the physiology of C. cuneata.

  18. RNA interference silencing of CHS greatly alters the growth pattern of apple (Malus x domestica).

    Science.gov (United States)

    Dare, Andrew P; Hellens, Roger P

    2013-08-01

    Plants produce a vast array of phenolic compounds which are essential for their survival on land. One major class of polyphenols are the flavonoids and their formation is dependent on the enzyme chalcone synthase (CHS). In a recent study we silenced the CHS genes of apple (Malus × domestica Borkh.) and observed a loss of pigmentation in the fruit skin, flowers and stems. More surprisingly, highly silenced lines were significantly reduced in size, with small leaves and shortened internode lengths. Chemical analysis also revealed that the transgenic shoots contained greatly reduced concentrations of flavonoids which are known to modulate auxin flow. An auxin transport study verified this, with an increased auxin transport in the CHS-silenced lines. Overall, these findings suggest that auxin transport in apple has adapted to take place in the presence of high endogenous concentrations of flavonoids. Removal of these compounds therefore results in abnormal auxin movement and a highly disrupted growth pattern.

  19. Unusual behavior of growing pollen tubes in the ovary of plum culture (Prunus domestica L.

    Directory of Open Access Journals (Sweden)

    Đorđević Milena

    2010-01-01

    Full Text Available Unusual behavior of growing pollen tubes in different combinations of pollination was observed in the ovary of the plum (Prunus domestica L. cv 'Čačanska Lepotica'. It primarily refers to several issues, i.e. the curling up of pollen tubes within the micropyle, the growth of two pollen tubes into the nucellus of an ovule, the occurrence of a bundle above the nucellar cap and fluorescence of the part of the embryo sac containing the egg apparatus. Upon the growth of pollen tubes into the nucellus of the ovule, subsequently penetrating pollen tubes form a bundle either above the micropyle entrance or above the nucellus. Branching and bending of pollen tubes by 180o upon their growth into the micropyle was also observed.

  20. Caffeoyl glucosides from Nandina domestica inhibit LPS-induced endothelial inflammatory responses.

    Science.gov (United States)

    Kulkarni, Roshan R; Lee, Wonhwa; Jang, Tae Su; Lee, JungIn; Kwak, Soyoung; Park, Mi Seon; Lee, Hyun-Shik; Bae, Jong-Sup; Na, MinKyun

    2015-11-15

    Endothelial dysfunction is a key pathological feature of many inflammatory diseases, including sepsis. In the present study, a new caffeoyl glucoside (1) and two known caffeoylated compounds (2 and 3) were isolated from the fruits of Nandina domestica Thunb. (Berberidaceae). The compounds were investigated for their effects against lipopolysaccharide (LPS)-mediated endothelial inflammatory responses. At 20 μM, 1 and 2 inhibited LPS-induced hyperpermeability, adhesion, and migration of leukocytes across a human endothelial cell monolayer in a dose-dependent manner suggesting that 1 and 2 may serve as potential scaffolds for the development of therapeutic agents to treat vascular inflammatory disorders. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Optimizing dehydration of apples Malus Domestica with fructo-oligosaccharide incorporation

    Directory of Open Access Journals (Sweden)

    Mariana Buranelo Egea

    2012-10-01

    Full Text Available The objective of the present study was to study the effect of the variables of the osmotic dehydration process on sliced Fuji apples (Malus domestica using a 2 x 3² factorial design. The variables studied in the apple slices were the pretreatment (blanching or acidification, the temperatures (30, 45 and 60ºC and the FOS concentration (40%, 50% and 60% m/v of the osmotic solution. There was no difference among the pretreatments for the water activity and titratable acidity. The slices pre-treated by the acidification presented less enzymatic browning (greatest luminosity L* value combined with a greater soluble solid contents (thus, this treatment was selected. Treatments T4 (45ºC and 40% m/v and T7 (60ºC and 40%m/v, using the acidification presented responses within the recommended standards and FOS were validated by the repetition.

  2. Antioxidant and antimicrobial properties of dried Portuguese apple variety (Malus domestica Borkh. cv Bravo de Esmolfe).

    Science.gov (United States)

    Pires, Tânia C S P; Dias, Maria Inês; Barros, Lillian; Alves, Maria José; Oliveira, M Beatriz P P; Santos-Buelga, Celestino; Ferreira, Isabel C F R

    2018-02-01

    Malus domestica Borkh apples are one of the most consumed fruits in the world, due to their sweetness and flavour. Herein, 'Bravo de Esmolfe' apple fruits were characterized regarding their nutritional value, chemical composition and bioactive properties. Besides nutrients, flavan-3-ols (i.e., epicatechin and B-type procyanidins) as also hydroxycinnamoyl-quinic acids and phloretin derivatives were identified in the samples. Extracts prepared from 'Bravo de Esmolfe' also proved to have antioxidant activity and antibacterial effects against Gram-positive bacteria, namely methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), Listeria monocytogenes and Enterococcus faecalis, and against the Gram-negative bacteria Escherichia coli, Escherichia coli (ESBL) (producing extended spectrum β-lactamases) and Morganella morganii. There is very little information about 'Bravo de Esmolfe' apple, so this study is important to inform consumers about an alternative source of nutritional and bioactive compounds. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Regional distribution of muscarinic acetylcholine receptors in the telencephalon of the pigeon (Columba livia f. domestica)

    International Nuclear Information System (INIS)

    Waechtler, K.

    1985-01-01

    The distribution of muscarinic acetylcholine receptors was studied autoradiographically in croystat sections of the pigeon telencephalon using 3 H-quinuclidinylbenzylate as a ligand. Highest receptor density was observed in the hyperstriatum ventrale, palaeostriatum augmentatum, septum, and parts of the archistriatum. In sites of known sensory input of neostriatum (field L) and ectostriatum low receptor binding was observed. Acetylcholinesterase distribution is in good agreement with the receptor picture only in the basal telencephalon. In the pallium differences in the pattern of these two components can be seen. (author)

  4. In silico analysis of the polygalacturonase inhibiting protein 1 from apple, Malus domestica.

    Science.gov (United States)

    Matsaunyane, Lerato Bt; Oelofse, Dean; Dubery, Ian A

    2015-03-11

    The Malus domestica polygalacturonase inhibiting protein 1 (MdPGIP1) gene, encoding the M. domestica polygalacturonase inhibiting protein 1 (MdPGIP1), was isolated from the Granny Smith apple cultivar (GenBank accession no. DQ185063). The gene was used to transform tobacco and potato for enhanced resistance against fungal diseases. Analysis of the MdPGIP1 nucleotide sequence revealed that the gene comprises 993 nucleotides that encode a 330 amino acid polypeptide. In silico characterization of the MdPGIP1 polypeptide revealed domains typical of PGIP proteins, which include a 24 amino acid putative signal peptide, a potential cleavage site [Alanine-Leucine-Serine (ALS)] for the signal peptide, a 238 amino acid leucine-rich repeat (LRR) domain, a 46 amino acid N-terminal domain and a 22 amino acid C-terminal domain. The hydropathic evaluation of MdPGIP1 indicated a repetitive hydrophobic motif in the LRR domain and a hydrophilic surface area consistent with a globular protein. The typical consensus glycosylation sequence of Asn-X-Ser/Thr was identified in MdPGIP1, indicating potential N-linked glycosylation of MdPGIP1. The molecular mass of non-glycosylated MdPGIP1 was calculated as 36.615 kDa and the theoretical isoelectric point as 6.98. Furthermore, the secondary and tertiary structure of MdPGIP1 was modelled, and revealed that MdPGIP1 is a curved and elongated molecule that contains sheet B1, sheet B2 and 310-helices on its LRR domain. The overall properties of the MdPGIP1 protein is similar to that of the prototypical Phaseolus vulgaris PGIP 2 (PvPGIP2), and the detected differences supported its use in biotechnological applications as an inhibitor of targeted fungal polygalacturonases (PGs).

  5. Efficacy and safety of Curcuma domestica extracts compared with ibuprofen in patients with knee osteoarthritis: a multicenter study

    Directory of Open Access Journals (Sweden)

    Kuptniratsaikul V

    2014-03-01

    Full Text Available Vilai Kuptniratsaikul,1 Piyapat Dajpratham,1 Wirat Taechaarpornkul,2 Montana Buntragulpoontawee,3 Pranee Lukkanapichonchut,4 Chirawan Chootip,5 Jittima Saengsuwan,6 Kesthamrong Tantayakom,7 Supphalak Laongpech8 1Department of Rehabilitation Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand; 2Sirindhorn National Medical Rehabilitation Center, Ministry of Public Health, Nonthaburi, Thailand; 3Department of Rehabilitation Medicine, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand; 4Department of Rehabilitation Medicine, Ratchaburi Hospital, Ministry of Public Health, Ratchaburi, Thailand; 5Department of Rehabilitation Medicine, Songkhla Hospital, Ministry of Public Health, Songkhla, Thailand; 6Department of Rehabilitation Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand; 7Department of Rehabilitation Medicine, Rajvithi Hospital, Ministry of Public Health, Bangkok, Thailand; 8Department of Rehabilitation Medicine, Vachira Phuket Hospital, Ministry of Public Health, Phuket, Thailand Objective: To determine the efficacy and safety of Curcuma domestica extracts in pain reduction and functional improvement. Methods: 367 primary knee osteoarthritis patients with a pain score of 5 or higher were randomized to receive ibuprofen 1,200 mg/day or C. domestica extracts 1,500 mg/day for 4 weeks. The main outcomes were Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC total, WOMAC pain, WOMAC stiffness, and WOMAC function scores. Adverse events (AEs were also recorded. Results: 185 and 182 patients were randomly assigned into C. domestica extracts and ibuprofen groups, respectively. The baseline characteristics were no different between groups. The mean of all WOMAC scores at weeks 0, 2, and 4 showed significant improvement when compared with the baseline in both groups. After using the noninferiority test, the mean difference (95% confidence interval of WOMAC total

  6. Inhibition of acetylcholinesterase activity in brain and behavioral analysis in adult rats after chronic administration of fenproporex.

    Science.gov (United States)

    Rezin, Gislaine T; Scaini, Giselli; Ferreira, Gabriela K; Cardoso, Mariane R; Gonçalves, Cinara L; Constantino, Larissa S; Deroza, Pedro F; Ghedim, Fernando V; Valvassori, Samira S; Resende, Wilson R; Quevedo, João; Zugno, Alexandra I; Streck, Emilio L

    2012-12-01

    Fenproporex is an amphetamine-based anorectic and it is rapidly converted in vivo into amphetamine. It elevates the levels of extracellular dopamine in the brain. Acetylcholinesterase is a regulatory enzyme which is involved in cholinergic synapses and may indirectly modulate the release of dopamine. Thus, we investigated whether the effects of chronic administration of fenproporex in adult rats alters acquisition and retention of avoidance memory and acetylcholinesterase activity. Adult male Wistar rats received repeated (14 days) intraperitoneal injection of vehicle or fenproporex (6.25, 12.5 or 25 mg/kg i.p.). For behavioral assessment, animals were submitted to inhibitory avoidance (IA) tasks and continuous multiple trials step-down inhibitory avoidance (CMIA). Acetylcholinesterase activity was measured in the prefrontal cortex, hippocampus, hypothalamus and striatum. The administration of fenproporex (6.25, 12.5 and 25 mg/kg) did not induce impairment in short and long-term IA or CMIA retention memory in rats. In addition, longer periods of exposure to fenproporex administration decreased acetylcholinesterase activity in prefrontal cortex and striatum of rats, but no alteration was verified in the hippocampus and hypothalamus. In conclusion, the present study showed that chronic fenproporex administration decreased acetylcholinesterase activity in the rat brain. However, longer periods of exposure to fenproporex did not produce impairment in short and long-term IA or CMIA retention memory in rats.

  7. Interleukin 6 modulates acetylcholinesterase activity of brain neurons; Effet de l`interleukine 6 sur l`activite de l`acetylcholinesterase des neurones centraux

    Energy Technology Data Exchange (ETDEWEB)

    Clarencon, D.; Multon, E.; Galonnier, M.; Estrade, M.; Fournier, C.; Mathieu, J.; Mestries, J.C.; Testylier, G.; Fatome, M.

    1995-12-31

    Classically, radiation injuries results in a peripheral inflammatory process, and we have previously observed an early systemic interleukin 6 (IL-6) release following whole-body irradiation. Besides, we have demonstrated an early decrease of rat or primate brain acetylcholinesterase (AChE) activity a gamma exposure. The object of the present study is to find possible IL-6 systemic effects on the brain AChE activity. We show that, though intravenous (i.v.) or intra-cerebro-ventricular (ICV) injection of IL-6 can induce a drop in rat brain AChE activity, this cytokine induces only a slight decrease of the AChE release in cultured brain cells. (author). 3 refs.

  8. Using multilevel systematic sampling to study apple fruit (Malus domestica Borkh.) quality and its variability at the orchard scale

    DEFF Research Database (Denmark)

    Martínez Vega, Mabel V.; Wulfsohn, Dvoralai; Clemmensen, Line Katrine Harder

    2013-01-01

    sample of ‘Granny Smith’ (Malus x domestica cv. ‘Granny Smith’) apples obtained from a 17 ha orchard based on a final sample of 74 fruit. Estimates of fruit marketable yield and fruit size distribution agreed well with packing house records. The estimated marketable yield was 356.6 ± 89.2 t compared...... recommended sample sizes to accurately describe the distribution of various quality variables of apples at the orchard scale....

  9. Mortality Due to Porcine Reproductive and Respiratory Syndrome Virus in Immunocompromised G?ttingen Minipigs (Sus scrofa domestica)

    OpenAIRE

    Pils, Marina C; Dreckmann, Karla; Jansson, Katharina; Glage, Silke; Held, Nadine; Sommer, Wiebke; L?nger, Florian; Avsar, Murat; Warnecke, Gregor; Bleich, Andr?

    2016-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) infection was diagnosed in 6 G?ttingen minipigs (Sus scrofa domestica) with severe interstitial pneumonia. The virus was defined as a North American (NA) subtype virus, which is common in the commercial pig population and might be derived from a widely used attenuated live-virus vaccine in Europe. The ORF5 sequence of the isolated PRRSV was 98% identical to the vaccine virus. The affected pigs were part of a lung transplantation mode...

  10. Chemical Composition and In Vitro Cytotoxic Activity of Essential Oil of Leaves of Malus domestica Growing in Western Himalaya (India)

    OpenAIRE

    Walia, Mayanka; Mann, Tavleen S.; Kumar, Dharmesh; Agnihotri, Vijai K.; Singh, Bikram

    2012-01-01

    Light pale-colored volatile oil was obtained from fresh leaves of Malus domestica tree, growing in Dhauladhar range of Himalaya (Himachal Pradesh, India), with characteristic eucalyptol dominant fragrance. The oil was found to be a complex mixture of mono-, sesqui-, di-terpenes, phenolics, and aliphatic hydrocarbons. Seventeen compounds accounting for nearly 95.3% of the oil were characterized with the help of capillary GC, GC-MS, and NMR. Major compounds of the oil were characterized as euca...

  11. Maize Lc transcription factor enhances biosynthesis of anthocyanins, distinct proanthocyanidins and phenylpropanoids in apple (Malus domestica Borkh.).

    Science.gov (United States)

    Li, Houhua; Flachowsky, Henryk; Fischer, Thilo C; Hanke, Magda-Viola; Forkmann, Gert; Treutter, Dieter; Schwab, Wilfried; Hoffmann, Thomas; Szankowski, Iris

    2007-10-01

    Flavonoids are a large family of polyphenolic compounds with manifold functions in plants. Present in a wide range of vegetables and fruits, flavonoids form an integral part of the human diet and confer multiple health benefits. Here, we report on metabolic engineering of the flavonoid biosynthetic pathways in apple (Malus domestica Borkh.) by overexpression of the maize (Zea mays L.) leaf colour (Lc) regulatory gene. The Lc gene was transferred into the M. domestica cultivar Holsteiner Cox via Agrobacterium tumefaciens-mediated transformation which resulted in enhanced anthocyanin accumulation in regenerated shoots. Five independent Lc lines were investigated for integration of Lc into the plant genome by Southern blot and PCR analyses. The Lc-transgenic lines contained one or two Lc gene copies and showed increased mRNA levels for phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), flavanone 3 beta-hydroxylase (FHT), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin reductases (LAR), anthocyanidin synthase (ANS) and anthocyanidin reductase (ANR). HPLC-DAD and LC-MS analyses revealed higher levels of the anthocyanin idaein (12-fold), the flavan 3-ol epicatechin (14-fold), and especially the isomeric catechin (41-fold), and some distinct dimeric proanthocyanidins (7 to 134-fold) in leaf tissues of Lc-transgenic lines. The levels of phenylpropanoids and their derivatives were only slightly increased. Thus, Lc overexpression in Malus domestica resulted in enhanced biosynthesis of specific flavonoid classes, which play important roles in both phytopathology and human health.

  12. Antioxidant and anti-acetylcholinesterase activities of extracts from Rapistrum rugosum in Tunisia

    Directory of Open Access Journals (Sweden)

    Omri Hichri Amel

    2017-07-01

    Full Text Available Objective: To investigate the antioxidant potential and anti-acetylcholinesterase activity of Rapistrum rugosum extracts. Methods: The crude, ethyl acetate, butanol and water extracts prepared from flowers, roots, stems and leaves of Rapistrum rugosum were tested at 1 mg/mL to determine their total polyphenol content, total flavonoid content and total condensed tannin content. Their antioxidant activity was assessed at different concentrations (0.0312, 0.0625, 0.1250, 0.25, 0.50 and 1.00 mg/ mL by using DPPH, ABTS, reducing power and β-carotene bleAChIng inhibition activity. Antiacetylcholinesterase activity was also determined. Results: The extract of leaves and stems had the highest total phenolic content [(110.45依0.03 mg gallic acid equivalent/g dry weight]. The ethyl acetate extract of flowers had the highest total flavonoid content [(24.62依0.13 mg quercetin equivalent/g dry weight]. The butanolic fraction of flowers had the highest total condensed tannin content [(317.85依0.01 mg catechin equivalent/g dry weight]. The crude extracts of flowers exhibited an interesting antioxidant activity for DPPH assay (93.00依0.01% at 1 mg/mL. The greatest acetylcholinesterase inhibitory activity (IC50=1.60 mg/mL was exhibited by the crude extracts from the flowers. Conclusions: The results demonstrated that Rapistrum rugosum contains active constituents which possess antioxidant and anti-acetylcholinesterase activities.

  13. Synthesis and Biological Evaluation of Novel Jatrorrhizine Derivatives with Amino Groups Linked at the 3-Position as Inhibitors of Acetylcholinesterase

    Directory of Open Access Journals (Sweden)

    Xiaofei Jiang

    2017-01-01

    Full Text Available Jatrorrhizine was considered as one of the active constituents of Coptis chinensis Franch. Herein, jatrorrhizine derivatives with substituted amino groups linked at the 3-position were designed, synthesized, and biologically evaluated as inhibitors of acetylcholinesterase. Jatrorrhizine derivatives inhibited the activity of acetylcholinesterase (AChE to a greater extent than the lead compound jatrorrhizine. All these jatrorrhizine derivatives were proved to be potent inhibitors of acetylcholinesterase (AChE with submicromolar IC50 values, but less sensitive to butyrylcholinesterase (BuChE, which suggests that these jatrorrhizine derivatives are selective for AChE/BuChE. Compound 3g gave the most potent inhibitor activity for AChE (IC50 = 0.301 μM, which is greater than the lead compound jatrorrhizine. All these results demonstrated that these jatrorrhizine derivatives are potential inhibitors for AChE.

  14. Acetylcholinesterase Inhibition and in Vitro and in Vivo Antioxidant Activities of Ganoderma lucidum Grown on Germinated Brown Rice

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    Beong Ou Lim

    2013-06-01

    Full Text Available In this study, the acetylcholinesterase inhibition and in vitro and in vivo antioxidant activities of Ganoderma lucidum grown on germinated brown rice (GLBR were evaluated. In antioxidant assays in vitro, GLBR was found to have strong metal chelating activity, DPPH, ABTS, hydroxyl and superoxide radical scavenging activity. Cell-based antioxidant methods were used, including lipid peroxidation on brain homogenate and AAPH-induced erythrocyte haemolysis. In antioxidant assays in vivo, mice were administered with GLBR and this significantly enhanced the activities of antioxidant enzymes in the mice sera, livers and brains. The amount of total phenolic and flavonoid compounds were 43.14 mg GAE/g and 13.36 mg CE/g dry mass, respectively. GLBR also exhibited acetylcholinesterase inhibitory activity. In addition, HPLC analyses of GLBR extract revealed the presence of different phenolic compounds. These findings demonstrate the remarkable potential of GLBR extract as valuable source of antioxidants which exhibit interesting acetylcholinesterase inhibitory activity.

  15. Theoretical NMR and conformational analysis of solvated oximes for organophosphates-inhibited acetylcholinesterase reactivation

    Science.gov (United States)

    da Silva, Jorge Alberto Valle; Modesto-Costa, Lucas; de Koning, Martijn C.; Borges, Itamar; França, Tanos Celmar Costa

    2018-01-01

    In this work, quaternary and non-quaternary oximes designed to bind at the peripheral site of acetylcholinesterase previously inhibited by organophosphates were investigated theoretically. Some of those oximes have a large number of degrees of freedom, thus requiring an accurate method to obtain molecular geometries. For this reason, the density functional theory (DFT) was employed to refine their molecular geometries after conformational analysis and to compare their 1H and 13C nuclear magnetic resonance (NMR) theoretical signals in gas-phase and in solvent. A good agreement with experimental data was achieved and the same theoretical approach was employed to obtain the geometries in water environment for further studies.

  16. Pharmacological evaluation of [11c]donepezil as tracer for visualization of acetylcholinesterase by PET

    International Nuclear Information System (INIS)

    Vos, F. de; Santens, P.; Vermeirsch, H.; Dewolf, I.; Dumont, F.; Slegers, G.; Dierckx, R.A.; Reuck, J. de

    2000-01-01

    Donepezil is a highly potent and selective reversible acetylcholinesterase inhibitor. [ 11 C]Donepezil is prepared by methylation with [ 11 C]CH 3 I of the corresponding 6'-O-desmethylprecursor. Tissue distribution in mice revealed a high uptake in brain and rapid clearance from the blood. Metabolization studies in mice indicated the formation of one 11 C-labeled polar metabolite that didn't penetrate the blood-brain barrier. Regional brain distribution in rabbits didn't reflect the measured achetylcholinesterase distribution in rabbit brain

  17. [Memantine as add-on medication to acetylcholinesterase inhibitor therapy for Alzheimer dementia].

    Science.gov (United States)

    Haussmann, R; Donix, M

    2017-01-01

    Currently available data indicate superior therapeutic effects of combination treatment for Alzheimer dementia with memantine and acetylcholine esterase inhibitors in certain clinical contexts. Out of five randomized, placebo-controlled, double-blind trials two showed superior therapeutic effects in comparison to monotherapy with acetylcholinesterase inhibitors regarding various domains. Recently published meta-analyses and cost-benefit analyses also showed positive results. Recently published German guidelines for dementia treatment also take these new data into account and recommend combination treatment in patients with severe dementia on stable donepezil medication. This article gives an overview of current evidence for combination therapy.

  18. Nanomaterials-Based Optical Techniques for the Detection of Acetylcholinesterase and Pesticides

    Directory of Open Access Journals (Sweden)

    Ning Xia

    2014-12-01

    Full Text Available The large amount of pesticide residues in the environment is a threat to global health by inhibition of acetylcholinesterase (AChE. Biosensors for inhibition of AChE have been thus developed for the detection of pesticides. In line with the rapid development of nanotechnology, nanomaterials have attracted great attention and have been intensively studied in biological analysis due to their unique chemical, physical and size properties. The aim of this review is to provide insight into nanomaterial-based optical techniques for the determination of AChE and pesticides, including colorimetric and fluorescent assays and surface plasmon resonance.

  19. Dihydroagarofuranoid Sesquiterpenes as Acetylcholinesterase Inhibitors from Celastraceae Plants: Maytenus disticha and Euonymus japonicus.

    Science.gov (United States)

    Alarcón, Julio; Cespedes, Carlos L; Muñoz, Evelyn; Balbontin, Cristian; Valdes, Francisco; Gutierrez, Margarita; Astudillo, Luis; Seigler, David S

    2015-12-02

    Natural cholinesterase inhibitors have been found in many biological sources. Nine compounds with agarofuran (epoxyeudesmane) skeletons were isolated from seeds and aerial parts of Maytenus disticha and Euonymus japonicus. The identification and structural elucidation of compounds were based on spectroscopic data analyses. All compounds had inhibitory acetylcholinesterase (AChE) activity. These natural compounds, which possessed mixed or uncompetitive mechanisms of inhibitory activity against AChE, may be considered as models for the design and development of new naturally occurring drugs for management strategies for neurodegenerative diseases. This is the first report of these chemical structures for seeds of M. disticha.

  20. Experimental transmission of Corynebacterium pseudotuberculosis biovar equi in horses by house flies

    Science.gov (United States)

    The route of Corynebacterium pseudotuberculosis infection in horses remains undetermined, but transmission by insects is suspected. Scientists from CMAVE and Auburn University investigated house flies (Musca domestica L.) as possible vectors. Three ponies were directly inoculated with C. pseudotuber...

  1. Flies (Calliphoridae, Muscidae and Beetles (Silphidae from Human Cadavers in Cali, Colombia

    Directory of Open Access Journals (Sweden)

    Barreto Mauricio

    2002-01-01

    Full Text Available Adult specimens of Cochliomyia macellaria, Chrysomya megacephala, Ch. rufifacies, Lucilia sp. (Calliphoridae, Musca domestica (Muscidae, Oxelytrum discicolle (Silphidae and Sarcophagidae were recovered from 12 human cadavers in Cali, Valle, Colombia. Information regarding these findings is presented.

  2. Identification of a trypsin-like site associated with acetylcholinesterase by affinity labelling with (/sup 3/H)diisopropyl fluorophosphate

    Energy Technology Data Exchange (ETDEWEB)

    Small, D.H.; Chubb, I.W.

    1988-07-01

    In addition to its ability to hydrolyze acetylcholine, purified eel acetylcholinesterase possesses a trypsin-like endopeptidase activity. The tryptic activity is associated with a serine residue at a site that is distinct from the esteratic site. To label both the esteratic and tryptic sites, the enzyme was incubated with the serine hydrolase inhibitor (/sup 3/H)diisopropyl fluorophosphate. This compound labelled the protein in a biphasic manner, with both slow and rapid labelling kinetics. The time course of the rapid phase was similar to the time course of inactivation of the esteratic activity. The time course of the slow phase was similar to the time course of inactivation of the tryptic activity. Labelling of the nonesteratic site was inhibited by the trypsin inhibitor N alpha-p-tosyl-L-lysine chloromethyl ketone. The total number of sites labelled by (/sup 3/H)diisopropyl fluorophosphate on eel acetylcholinesterase was 2.6 mol/280,000 g protein, whereas the number of tryptic sites was less (0.52 mol/280,000 g). The results suggest that a subpopulation of acetylcholinesterase molecules may possess tryptic activity. Extensive chromatography of the purified enzyme by ion-exchange and gel filtration failed to separate the labelled tryptic component from acetylcholinesterase. On sodium dodecyl sulfate-polyacrylamide gels, the labelled tryptic component comigrated with a polypeptide of 50,000 molecular weight, which is a major proteolytic digestion product derived from the intact acetylcholinesterase monomer. Because of its localization in many noncholinergic peptide-containing cells, acetylcholinesterase could act as a neuropeptide processing enzyme in these cells.

  3. In vitro toxicity, antiplatelet and acetylcholinesterase inhibition of Buddleja thyrsoides Lam. leaves.

    Science.gov (United States)

    Mahlke, Janaína Dorneles; Boligon, Aline Augusti; Machado, Michel Mansur; Athayde, Margareth Linde

    2012-01-01

    Alzheimer's disease (AD) is a neurodegenerative disorder resulting in impaired memory and behaviour of remarkable socio-economic impact. A decrease in cholinergic activity is a key event in the biochemical of AD. Buddleja thyrsoides is a plant widely distributed in Southern parts of South America. In Brazilian traditional medicine, the infusion of its leaves and flowers is used for the treatment of bronchitis and cough. Crude ethanolic (70%) extract and fractions (dichloromethane, ethyl acetate and n-butanolic) were investigated regarding their toxicities in vitro and antiplatelet action. The enzyme acetylcholinesterase inhibition was evaluated to study the crude extract. The crude extract and fractions were evaluated by means of Brine Shrimp Lethality test and they showed low activities with LC(50) values 1698, 2818, 2187 and 3672 µg mL(-1) for dichloromethane, ethyl acetate, n-butanolic fractions and crude extract, respectively. Buddleja thyrsoides presented great antiplatelet action. The IC(50) values obtained for crude extract and dichloromethane, ethyl acetate and n-butanolic fractions were 361.29, 354.23, 368.75 and 344.30, respectively, while the IC(50) for the standard AAS was 257.01 µg mL(-1). The crude extract showed an inhibition of 22.8% of the acetylcholinesterase enzyme in 24 h.

  4. Is acetylcholinesterase a biomarker of susceptibility in Daphnia magna (Crustacea, Cladocera) after deltamethrin exposure?

    Science.gov (United States)

    Toumi, Héla; Boumaiza, Moncef; Millet, Maurice; Radetski, Claudemir Marcos; Felten, Vincent; Férard, Jean François

    2015-02-01

    In the present study, we explored the possibility of using the acetylcholinesterase (AChE) as a biomarker after deltamethrin (pyrethroid insecticide) exposure with three strains of the cladoceran Daphnia magna. Four calculated time-weighted deltamethrin concentrations (20.1, 40.3, 80.6 and 161.3 ng L(-1)) were compared against control acetylcholinesterase activity. Our results showed that after 48 h of deltamethrin exposure, all treatments induced a significant decrease of AChE activities whatever the three considered strains. However, diverse responses were registered in terms of lowest observed effect concentrations (LOEC: 80.6 ng L(-1) for strain 1 and 20.1 ng L(-1) for strains 2 and 3) revealing differences in sensitivity among the three tested strains of D. magna. Our results suggest that after deltamethrin exposure, the AChE activity responses can be also used as a biomarker of susceptibility (i.e., variation of strain specific response). Moreover, our results show that strain 1 is the less sensitive in terms of IC50-48 h of AChE, whereas it became the most sensitive when considering the EC50-48 h estimated in the standard ecotoxicity test. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Screening of acetylcholinesterase inhibitors in marine organisms from the Caribbean Sea.

    Science.gov (United States)

    Castellanos, Fabio; Amaya-García, Fabián; Tello, Edisson; Ramos, Freddy Alejandro; Umaña, Adriana; Puyana, Mónica; Resende, Jackson A L C; Castellanos, Leonardo

    2018-06-04

    The acetylcholinesterase inhibitory activity of 89 organic extracts from marine organisms was evaluated through a TLC bioautography methodology. Extracts from soft corals (Eunicea and Plexaura) were the most active compared with extracts from sponges. The bioguided chemical study of the most active extract, obtained from Pseudoplexaura porosa, led to the isolation of a diterpene with spectroscopic properties consistent to those published to the cembrane Steylolide. However, further analysis by X-ray diffraction indicated that the compound was the 14-acetoxycrassine (1), correcting the structure reported to the Styelolide. Additionally, the acetylcholinesterase inhibitory activity of fourteen cembranoids (2-15) isolated from soft corals Eunicea knighti and Pseudoplexaura flagellosa was evaluated. Cembranoids 2, 3 and 4 were the most active compounds in the TLC bioassay. Then, the most promising cembranoids, 14-acetoxycrassine (1) and asperdiol (2), were tested quantitatively and they exhibited IC 50 values of 1.40 ± 0.113 and 0.358 ± 0.130 μM, respectively.

  6. Acetylcholinesterase-Inhibiting Activity of Salicylanilide N-Alkylcarbamates and Their Molecular Docking

    Directory of Open Access Journals (Sweden)

    Josef Jampilek

    2012-08-01

    Full Text Available A series of twenty-five novel salicylanilide N-alkylcarbamates were investigated as potential acetylcholinesterase inhibitors. The compounds were tested for their ability to inhibit acetylcholinesterase (AChE from electric eel (Electrophorus electricus L.. Experimental lipophilicity was determined, and the structure-activity relationships are discussed. The mode of binding in the active site of AChE was investigated by molecular docking. All the discussed compounds expressed significantly higher AChE inhibitory activity than rivastigmine and slightly lower than galanthamine. Disubstitution by chlorine in C'(3,4 of the aniline ring and the optimal length of hexyl-undecyl alkyl chains in the carbamate moiety provided the most active AChE inhibitors. Monochlorination in C'(4 exhibited slightly more effective AChE inhibitors than in C'(3. Generally it can be stated that compounds with higher lipophilicity showed higher inhibition, and the activity of the compounds is strongly dependent on the length of the N-alkyl chain.

  7. Global loss of acetylcholinesterase activity with mitochondrial complexes inhibition and inflammation in brain of hypercholesterolemic mice.

    Science.gov (United States)

    Paul, Rajib; Borah, Anupom

    2017-12-20

    There exists an intricate relationship between hypercholesterolemia (elevated plasma cholesterol) and brain functions. The present study aims to understand the impact of hypercholesterolemia on pathological consequences in mouse brain. A chronic mouse model of hypercholesterolemia was induced by giving high-cholesterol diet for 12 weeks. The hypercholesterolemic mice developed cognitive impairment as evident from object recognition memory test. Cholesterol accumulation was observed in four discrete brain regions, such as cortex, striatum, hippocampus and substantia nigra along with significantly damaged blood-brain barrier by hypercholesterolemia. The crucial finding is the loss of acetylcholinesterase activity with mitochondrial dysfunction globally in the brain of hypercholesterolemic mice, which is related to the levels of cholesterol. Moreover, the levels of hydroxyl radical were elevated in the regions of brain where the activity of mitochondrial complexes was found to be reduced. Intriguingly, elevations of inflammatory stress markers in the cholesterol-rich brain regions were observed. As cognitive impairment, diminished brain acetylcholinesterase activity, mitochondrial dysfunctions, and inflammation are the prima facie pathologies of neurodegenerative diseases, the findings impose hypercholesterolemia as potential risk factor towards brain dysfunction.

  8. A reversed-phase compatible thin-layer chromatography autography for the detection of acetylcholinesterase inhibitors.

    Science.gov (United States)

    Ramallo, I Ayelen; García, Paula; Furlan, Ricardo L E

    2015-11-01

    A dual readout autographic assay to detect acetylcholinesterase inhibitors present in complex matrices adsorbed on reversed-phase or normal-phase thin-layer chromatography plates is described. Enzyme gel entrapment with an amphiphilic copolymer was used for assay development. The effects of substrate and enzyme concentrations, pH, incubation time, and incubation temperature on the sensitivity and the detection limit of the assay were evaluated. Experimental design and response surface methodology were used to optimize conditions with a minimum number of experiments. The assay allowed the detection of 0.01% w/w of physostigmine in both a spiked Sonchus oleraceus L. extract chromatographed on normal phase and a spiked Pimenta racemosa (Mill.) J.W. Moore leaf essential oil chromatographed on reversed phase. Finally, the reversed-phase thin-layer chromatography assay was applied to reveal the presence of an inhibitor in the Cymbopogon citratus (DC.) Stapf essential oil. The developed assay is able to detect acetylcholinesterase inhibitors present in complex matrixes that were chromatographed in normal phase or reversed-phase thin-layer chromatography. The detection limit for physostigmine on both normal and reversed phase was of 1×10(-4) μg. The results can be read by a change in color and/or a change in fluorescence. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. An acetylcholinesterase biosensor based on graphene-gold nanocomposite and calcined layered double hydroxide.

    Science.gov (United States)

    Zhai, Chen; Guo, Yemin; Sun, Xia; Zheng, Yuhe; Wang, Xiangyou

    2014-05-10

    In this study, a novel acetylcholinesterase-based biosensor was fabricated. Acetylcholinesterase (AChE) was immobilized onto a glassy carbon electrode (GCE) with the aid of Cu-Mg-Al calcined layered double hydroxide (CLDH). CLDH can provide a bigger effective surface area for AChE loading, which could improve the precision and stability of AChE biosensor. However, the poor electroconductibility of CLDHs could lead to the low sensitivity of AChE biosensor. In order to effectively compensate the disadvantages of CLDHs, graphene-gold nanocomposites were used for improving the electron transfer rate. Thus, the graphene-gold nanocomposite (GN-AuNPs) was firstly modified onto the GCE, and then the prepared CLDH-AChE composite was immobilized onto the modified GCE to construct a sensitive AChE biosensor for pesticides detection. Relevant parameters were studied in detail and optimized, including the pH of the acetylthiocholine chloride (ATCl) solution, the amount of AChE immobilized on the biosensor and the inhibition time governing the analytical performance of the biosensor. The biosensor detected chlorpyrifos at concentrations ranging from 0.05 to 150μg/L. The detection limit for chlorpyrifos was 0.05μg/L. Copyright © 2014 Elsevier Inc. All rights reserved.

  10. Novel and viable acetylcholinesterase target site for developing effective and environmentally safe insecticides.

    Science.gov (United States)

    Pang, Yuan-Ping; Brimijoin, Stephen; Ragsdale, David W; Zhu, Kun Yan; Suranyi, Robert

    2012-04-01

    Insect pests are responsible for human suffering and financial losses worldwide. New and environmentally safe insecticides are urgently needed to cope with these serious problems. Resistance to current insecticides has resulted in a resurgence of insect pests, and growing concerns about insecticide toxicity to humans discourage the use of insecticides for pest control. The small market for insecticides has hampered insecticide development; however, advances in genomics and structural genomics offer new opportunities to develop insecticides that are less dependent on the insecticide market. This review summarizes the literature data that support the hypothesis that an insect-specific cysteine residue located at the opening of the acetylcholinesterase active site is a promising target site for developing new insecticides with reduced off-target toxicity and low propensity for insect resistance. These data are used to discuss the differences between targeting the insect-specific cysteine residue and targeting the ubiquitous catalytic serine residue of acetylcholinesterase from the perspective of reducing off-target toxicity and insect resistance. Also discussed is the prospect of developing cysteine-targeting anticholinesterases as effective and environmentally safe insecticides for control of disease vectors, crop damage, and residential insect pests within the financial confines of the present insecticide market.

  11. Virtual Screening of Acetylcholinesterase Inhibitors Using the Lipinski’s Rule of Five and ZINC Databank

    Directory of Open Access Journals (Sweden)

    Pablo Andrei Nogara

    2015-01-01

    Full Text Available Alzheimer’s disease (AD is a progressive and neurodegenerative pathology that can affect people over 65 years of age. It causes several complications, such as behavioral changes, language deficits, depression, and memory impairments. One of the methods used to treat AD is the increase of acetylcholine (ACh in the brain by using acetylcholinesterase inhibitors (AChEIs. In this study, we used the ZINC databank and the Lipinski’s rule of five to perform a virtual screening and a molecular docking (using Auto Dock Vina 1.1.1 aiming to select possible compounds that have quaternary ammonium atom able to inhibit acetylcholinesterase (AChE activity. The molecules were obtained by screening and further in vitro assays were performed to analyze the most potent inhibitors through the IC50 value and also to describe the interaction models between inhibitors and enzyme by molecular docking. The results showed that compound D inhibited AChE activity from different vertebrate sources and butyrylcholinesterase (BChE from Equus ferus (EfBChE, with IC50 ranging from 1.69 ± 0.46 to 5.64 ± 2.47 µM. Compound D interacted with the peripheral anionic subsite in both enzymes, blocking substrate entrance to the active site. In contrast, compound C had higher specificity as inhibitor of EfBChE. In conclusion, the screening was effective in finding inhibitors of AChE and BuChE from different organisms.

  12. Selective radiolabeling and isolation of the hydrophobic membrane-binding domain of human erythrocyte acetylcholinesterase

    International Nuclear Information System (INIS)

    Roberts, W.L.; Rosenberry, T.L.

    1986-01-01

    The hydrophobic, membrane-binding domain of purified human erythrocyte acetylcholinesterase was labeled with the photoactivated reagent 3-(trifluoromethyl)-3-(m-[ 125 I]iodophenyl)diazirine. The radiolabel was incorporated when the enzyme was prepared in detergent-free aggregates, in detergent micelles, or in phospholipid liposomes, but the highest percentage of labeling occurred in the detergent-free aggregates. Papain digestion of the enzyme released the hydrophobic domain, and polyacrylamide gel electrophoresis in sodium dodecyl sulfate or gel exclusion chromatography demonstrated that the label was localized exclusively in the cleaved hydrophobic domain fragment. This fragment was purified in a three-step procedure. Digestion was conducted with papain attached to Sepharose CL-4B, and the supernatant was adsorbed to acridinium affinity resin to remove the hydrophilic enzyme fragment. The nonretained fragment associated with Triton X-100 micelles was then chromatographed on Sepharose CL-6B, and finally detergent was removed by chromatography on Sephadex LH-60 in an ethanol-formic acid solvent. The fragment exhibited an apparent molecular weight of 3100 on the Sephadex LH-60 column when compared with peptide standards. However, amino acid analysis of the purified fragment revealed only 1 mol each of histidine and glycine per mole of fragment in contrast to the 25-30 mole of amino acids expected on the basis of the molecular weight estimate. This result suggests a novel non-amino acid structure for the hydrophobic domain of human erythrocyte acetylcholinesterase

  13. Transcriptomic analysis of Prunus domestica undergoing hypersensitive response to plum pox virus infection.

    Science.gov (United States)

    Rodamilans, Bernardo; San León, David; Mühlberger, Louisa; Candresse, Thierry; Neumüller, Michael; Oliveros, Juan Carlos; García, Juan Antonio

    2014-01-01

    Plum pox virus (PPV) infects Prunus trees around the globe, posing serious fruit production problems and causing severe economic losses. One variety of Prunus domestica, named 'Jojo', develops a hypersensitive response to viral infection. Here we compared infected and non-infected samples using next-generation RNA sequencing to characterize the genetic complexity of the viral population in infected samples and to identify genes involved in development of the resistance response. Analysis of viral reads from the infected samples allowed reconstruction of a PPV-D consensus sequence. De novo reconstruction showed a second viral isolate of the PPV-Rec strain. RNA-seq analysis of PPV-infected 'Jojo' trees identified 2,234 and 786 unigenes that were significantly up- or downregulated, respectively (false discovery rate; FDR≤0.01). Expression of genes associated with defense was generally enhanced, while expression of those related to photosynthesis was repressed. Of the total of 3,020 differentially expressed unigenes, 154 were characterized as potential resistance genes, 10 of which were included in the NBS-LRR type. Given their possible role in plant defense, we selected 75 additional unigenes as candidates for further study. The combination of next-generation sequencing and a Prunus variety that develops a hypersensitive response to PPV infection provided an opportunity to study the factors involved in this plant defense mechanism. Transcriptomic analysis presented an overview of the changes that occur during PPV infection as a whole, and identified candidates suitable for further functional characterization.

  14. Effect of irradiance and spectral composition of radiation on in vitro shoot proliferation in Malus domestica

    International Nuclear Information System (INIS)

    Noè, N.; Eccher, T.; Bonini, L.

    1997-01-01

    Four clones of Malus domestica cv. Golden Delicious - namely Smoothee, Crielaard, Reinders and Golden B - were cultured in vitro from single-node microcuttings placed on solid medium under irradiance (PPFD) of 50 micromol m -2 s -1 . After 9 months an average shoot proliferation of 5.3 was achieved; Crielaard showed the highest rate (7.1), followed Golden B (5.4), Smoothee and Reinders (4.4). Proliferating shoots were then exposed to higher PPFD (80 micromol m -2 s -1 ) and different spectral composition of radiation using PMMA-B and PMMA-R/FR filters. High PPFD decreased the average proliferation rate to 4.5, in particular in Crielaard and Golden B, while it increased proliferation in Reinders. When a PMMA-R/FR filter was interposed, the mean proliferation rate slightly increased. PMMA-B filters decreased the overall proliferation rate to 3.0; only in Crielaard it was increased, but shoots were very small. Thus PPFD and spectral composition influenced in vitro shoot proliferation and growth and the responses were different among the clones. (author)

  15. A three-dimensional stereotaxic atlas of the gray short-tailed opossum (Monodelphis domestica) brain.

    Science.gov (United States)

    Majka, Piotr; Chlodzinska, Natalia; Turlejski, Krzysztof; Banasik, Tomasz; Djavadian, Ruzanna L; Węglarz, Władysław P; Wójcik, Daniel K

    2018-05-01

    The gray short-tailed opossum (Monodelphis domestica) is a small marsupial gaining recognition as a laboratory animal in biomedical research. Despite numerous studies on opossum neuroanatomy, a consistent and comprehensive neuroanatomical reference for this species is still missing. Here we present the first three-dimensional, multimodal atlas of the Monodelphis opossum brain. It is based on four complementary imaging modalities: high resolution ex vivo magnetic resonance images, micro-computed tomography scans of the cranium, images of the face of the cutting block, and series of sections stained with the Nissl method and for myelinated fibers. Individual imaging modalities were reconstructed into a three-dimensional form and then registered to the MR image by means of affine and deformable registration routines. Based on a superimposition of the 3D images, 113 anatomical structures were demarcated and the volumes of individual regions were measured. The stereotaxic coordinate system was defined using a set of cranial landmarks: interaural line, bregma, and lambda, which allows for easy expression of any location within the brain with respect to the skull. The atlas is released under the Creative Commons license and available through various digital atlasing web services.

  16. Molecular survey on zoonotic tick-borne bacteria and chlamydiae in feral pigeons (Columba livia domestica).

    Science.gov (United States)

    Ebani, Valentina Virginia; Bertelloni, Fabrizio; Mani, Paolo

    2016-04-01

    To determine the presence of zoonotic tick-borne bacteria in feral pigeons (Columba livia domestica) from urban areas. Spleen samples from 84 feral pigeons, found dead with traumatic injuries in urban areas, were examined by PCR to detect DNA of Anaplasma phagocytophilum, Bartonella spp., Borrelia burgdorferi sensu lato, Coxiella burnetii, Rickettsia spp., and Chlamydophila spp. Twenty (23.8%) pigeons were infected by tick-borne agents, in particular 2 (2.38%) animals resulted positive for Bartonella spp., 5 (5.95%) for C. burnetii, 5 (5.95%) for Rickettsia spp., 13 (15.47%) for B. burgdorferi sensu lato. All birds scored negative for A. phagocytophilum. Moreover, 17 (20.23%) pigeons were positive for Chlamydophila spp. and among them 10 (11.9%) for Chlamydophila psittaci. Mixed infections by two or three agents were detected in 8 (9.52%) animals. Feral pigeons living in urban and periurban areas are a hazard for the human health as source of several pathogens. The obtained results confirm pigeons as reservoirs of chlamydial agents and suggest that they may be involved in the epidemiology of zoonotic tick-borne infections too. Copyright © 2016 Hainan Medical College. Production and hosting by Elsevier B.V. All rights reserved.

  17. Homing pigeons ( Columba livia f. domestica) can use magnetic cues for locating food

    Science.gov (United States)

    Thalau, Peter; Holtkamp-Rötzler, Elke; Fleissner, Gerta; Wiltschko, Wolfgang

    2007-10-01

    An experimental group of homing pigeons ( Columba livia f. domestica) learned to associate food with a magnetic anomaly produced by bar magnets that were fixed to the bowl in which they received their daily food ration in their home loft; the control group lacked this experience. Both groups were trained to search for two hidden food depots in a rectangular sand-filled arena without obvious visual cues; for the experimental birds, these depots were also marked with three 1.15 × 106 μT bar magnets. During the tests, there were two food depots, one marked with the magnets, the other unmarked; their position within the arena was changed from test to test. The experimental birds searched within 10 cm of the magnetically marked depot in 49% of the test sessions, whereas the control birds searched there in only 11% of the sessions. Both groups searched near the control depot in 11 and 13% of the sessions, respectively. The significant preference of the magnetically marked food depot by the experimental birds shows that homing pigeons cannot only detect a magnetic anomaly but can also use it as a cue for locating hidden food in an open arena.

  18. Transcriptomic analysis of Prunus domestica undergoing hypersensitive response to plum pox virus infection.

    Directory of Open Access Journals (Sweden)

    Bernardo Rodamilans

    Full Text Available Plum pox virus (PPV infects Prunus trees around the globe, posing serious fruit production problems and causing severe economic losses. One variety of Prunus domestica, named 'Jojo', develops a hypersensitive response to viral infection. Here we compared infected and non-infected samples using next-generation RNA sequencing to characterize the genetic complexity of the viral population in infected samples and to identify genes involved in development of the resistance response. Analysis of viral reads from the infected samples allowed reconstruction of a PPV-D consensus sequence. De novo reconstruction showed a second viral isolate of the PPV-Rec strain. RNA-seq analysis of PPV-infected 'Jojo' trees identified 2,234 and 786 unigenes that were significantly up- or downregulated, respectively (false discovery rate; FDR≤0.01. Expression of genes associated with defense was generally enhanced, while expression of those related to photosynthesis was repressed. Of the total of 3,020 differentially expressed unigenes, 154 were characterized as potential resistance genes, 10 of which were included in the NBS-LRR type. Given their possible role in plant defense, we selected 75 additional unigenes as candidates for further study. The combination of next-generation sequencing and a Prunus variety that develops a hypersensitive response to PPV infection provided an opportunity to study the factors involved in this plant defense mechanism. Transcriptomic analysis presented an overview of the changes that occur during PPV infection as a whole, and identified candidates suitable for further functional characterization.

  19. Phenotypic changes associated with RNA interference silencing of chalcone synthase in apple (Malus × domestica).

    Science.gov (United States)

    Dare, Andrew P; Tomes, Sumathi; Jones, Midori; McGhie, Tony K; Stevenson, David E; Johnson, Ross A; Greenwood, David R; Hellens, Roger P

    2013-05-01

    We have identified in apple (Malus × domestica) three chalcone synthase (CHS) genes. In order to understand the functional redundancy of this gene family RNA interference knockout lines were generated where all three of these genes were down-regulated. These lines had no detectable anthocyanins and radically reduced concentrations of dihydrochalcones and flavonoids. Surprisingly, down-regulation of CHS also led to major changes in plant development, resulting in plants with shortened internode lengths, smaller leaves and a greatly reduced growth rate. Microscopic analysis revealed that these phenotypic changes extended down to the cellular level, with CHS-silenced lines showing aberrant cellular organisation in the leaves. Fruit collected from one CHS-silenced line was smaller than the 'Royal Gala' controls, lacked flavonoids in the skin and flesh and also had changes in cell morphology. Auxin transport experiments showed increased rates of auxin transport in a CHS-silenced line compared with the 'Royal Gala' control. As flavonoids are well known to be key modulators of auxin transport, we hypothesise that the removal of almost all flavonoids from the plant by CHS silencing creates a vastly altered environment for auxin transport to occur and results in the observed changes in growth and development. © 2013 The Authors The Plant Journal © 2013 Blackwell Publishing Ltd.

  20. Involvement of Auxin and Brassinosteroid in Dwarfism of Autotetraploid Apple (Malus × domestica).

    Science.gov (United States)

    Ma, Yue; Xue, Hao; Zhang, Lei; Zhang, Feng; Ou, Chunqing; Wang, Feng; Zhang, Zhihong

    2016-05-24

    The plant height is an important trait in fruit tree. However, the molecular mechanism on dwarfism is still poorly understood. We found that colchicine-induced autotetraploid apple plants (Malus × domestica) exhibited a dwarf phenotype. The vertical length of cortical parenchyma cells was shorter in autotetraploids than in diploids, by observing paraffin sections. Hormone levels of indoleacetic acid (IAA) and brassinosteroid (BR) were significantly decreased in 3- and 5-year-old autotetraploid plants. Digital gene expression (DGE) analysis showed that the differentially expressed genes were mainly involved in IAA and BR pathways. microRNA390 was significantly upregulated according to microarray analysis. Exogenous application of IAA and BR promoted stem elongation of both apple plants grown in medium. The results show that dwarfing in autotetraploid apple plants is most likely regulated by IAA and BR. The dwarf phenotype of autotetraploid apple plants could be due to accumulation of miR390 after genome doubling, leading to upregulation of apple trans-acting short-interfering RNA 3 (MdTAS3) expression, which in turn downregulates the expression of MdARF3. Overall, this leads to partial interruption of the IAA and BR signal transduction pathway. Our study provides important insights into the molecular mechanisms underlying dwarfism in autopolyploid apple plants.

  1. Characterization of three chalcone synthase-like genes from apple (Malus x domestica Borkh.).

    Science.gov (United States)

    Yahyaa, Mosaab; Ali, Samah; Davidovich-Rikanati, Rachel; Ibdah, Muhammad; Shachtier, Alona; Eyal, Yoram; Lewinsohn, Efraim; Ibdah, Mwafaq

    2017-08-01

    Apple (Malus x domestica Brokh.) is a widely cultivated deciduous tree species of significant economic importance. Apple leaves accumulate high levels of flavonoids and dihydrochalcones, and their formation is dependent on enzymes of the chalcone synthase family. Three CHS genes were cloned from apple leaves and expressed in Escherichia coli. The encoded recombinant enzymes were purified and functionally characterized. In-vitro activity assays indicated that MdCHS1, MdCHS2 and MdCHS3 code for proteins exhibiting polyketide synthase activity that accepted either p-dihydrocoumaroyl-CoA, p-coumaroyl-CoA, or cinnamoyl-CoA as starter CoA substrates in the presence of malonyl-CoA, leading to production of phloretin, naringenin chalcone, and pinocembrin chalcone. MdCHS3 coded a chalcone-dihydrochalcone synthase enzyme with narrower substrate specificity than the previous ones. The apparent Km values of MdCHS3 for p-dihydrocoumaryl-CoA and p-coumaryl-CoA were both 5.0 μM. Expression analyses of MdCHS genes varied according to tissue type. MdCHS1, MdCHS2 and MdCHS3 expression levels were associated with the levels of phloretin accumulate in the respective tissues. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Trehalose 6-phosphate signal is closely related to sorbitol in apple (Malus domestica Borkh. cv. Gala)

    Science.gov (United States)

    Zhang, Wen; Lunn, John E.; Feil, Regina; Wang, Yufei; Zhao, Jingjing; Tao, Hongxia; Zhao, Zhengyang

    2017-01-01

    ABSTRACT Trehalose-6-phosphate (Tre6P) is a precursor of trehalose, which is widespread in nature and greatly influences plant growth and development. Tre6P acts as a signal of carbon availability in many plants, but little is known about the function of Tre6P in rosaceous plants, which have specific sorbitol biosynthesis and transportation pathways. In the present study, Tre6P levels and Sorbitol:Tre6P ratios were analyzed in apple (Malus domestica, Borkh. cv. Gala). Tre6P levels were positively correlated with sorbitol content but negatively correlated with sucrose, glucose, and fructose content in developing fruit. However, under sorbitol-limited conditions, Tre6P levels were positively correlated with both sorbitol and sucrose. In the presence of different exogenous sugar supply, Tre6P levels increased corresponding with sorbitol, but this was not the case with sucrose. In addition, Tre6P content and sorbitol:Tre6P ratios were more highly correlated with ADP-glucose levels under sorbitol-limited conditions and fruit development stages, respectively. These results suggest that Tre6P is more closely related to sorbitol than other soluble sugars and has an important role in influencing carbon metabolism in apple. PMID:28069587

  3. Future bloom and blossom frost risk for Malus domestica considering climate model and impact model uncertainties.

    Science.gov (United States)

    Hoffmann, Holger; Rath, Thomas

    2013-01-01

    The future bloom and risk of blossom frosts for Malus domestica were projected using regional climate realizations and phenological ( = impact) models. As climate impact projections are susceptible to uncertainties of climate and impact models and model concatenation, the significant horizon of the climate impact signal was analyzed by applying 7 impact models, including two new developments, on 13 climate realizations of the IPCC emission scenario A1B. Advancement of phenophases and a decrease in blossom frost risk for Lower Saxony (Germany) for early and late ripeners was determined by six out of seven phenological models. Single model/single grid point time series of bloom showed significant trends by 2021-2050 compared to 1971-2000, whereas the joint signal of all climate and impact models did not stabilize until 2043. Regarding blossom frost risk, joint projection variability exceeded the projected signal. Thus, blossom frost risk cannot be stated to be lower by the end of the 21st century despite a negative trend. As a consequence it is however unlikely to increase. Uncertainty of temperature, blooming date and blossom frost risk projection reached a minimum at 2078-2087. The projected phenophases advanced by 5.5 d K(-1), showing partial compensation of delayed fulfillment of the winter chill requirement and faster completion of the following forcing phase in spring. Finally, phenological model performance was improved by considering the length of day.

  4. Trehalose 6-phosphate signal is closely related to sorbitol in apple (Malus domestica Borkh. cv. Gala

    Directory of Open Access Journals (Sweden)

    Wen Zhang

    2017-02-01

    Full Text Available Trehalose-6-phosphate (Tre6P is a precursor of trehalose, which is widespread in nature and greatly influences plant growth and development. Tre6P acts as a signal of carbon availability in many plants, but little is known about the function of Tre6P in rosaceous plants, which have specific sorbitol biosynthesis and transportation pathways. In the present study, Tre6P levels and Sorbitol:Tre6P ratios were analyzed in apple (Malus domestica, Borkh. cv. Gala. Tre6P levels were positively correlated with sorbitol content but negatively correlated with sucrose, glucose, and fructose content in developing fruit. However, under sorbitol-limited conditions, Tre6P levels were positively correlated with both sorbitol and sucrose. In the presence of different exogenous sugar supply, Tre6P levels increased corresponding with sorbitol, but this was not the case with sucrose. In addition, Tre6P content and sorbitol:Tre6P ratios were more highly correlated with ADP-glucose levels under sorbitol-limited conditions and fruit development stages, respectively. These results suggest that Tre6P is more closely related to sorbitol than other soluble sugars and has an important role in influencing carbon metabolism in apple.

  5. Peripheral site ligand conjugation to a non-quaternary oxime enhances reactivation of nerve agent-inhibited human acetylcholinesterase

    NARCIS (Netherlands)

    Koning, M.C. de; Grol, M. van; Noort, D.

    2011-01-01

    Commonly employed pyridinium-oxime (charged) reactivators of nerve agent inhibited acetylcholinesterase (AChE) do not readily pass the blood brain barrier (BBB) because of the presence of charge(s). Conversely, non-ionic oxime reactivators often suffer from a lack of reactivating potency due to a

  6. High-performance liquid chromatography-mass spectrometry-based acetylcholinesterase assay for the screening of inhibitors in natural extracts

    NARCIS (Netherlands)

    de Jong, C.F.; Derks, R.J.E.; Bruyneel, B.; Niessen, W.M.A.; Irth, H.

    2006-01-01

    The present paper describes a High-performance liquid chromatography-mass spectrometry (LC-MS) methodology for the screening of acetylcholinesterase (AChE) inhibitors in natural extracts. AChE activity of sample components is monitored by a post-column biochemical assay that is based on the

  7. Acetylcholinesterase of Rhipicephalus (Boophilus) microplus and Phlebotomus papatasi: Gene identification, expression, and biochemical properties of recombinant proteins

    Science.gov (United States)

    Rhipicephalus (Boophilus) microplus (Bm) ticks are vectors of bovine babesiosis and anaplasmosis. Tick resistance to organophosphate (OP) acaricide involves acetylcholinesterase (AChE) insensitivity to OP and metabolic detoxification. Sequencing and in vitro expression of Bm genes encoding AChE allo...

  8. Acetylcholinesterases of Rhipicephalus (Boophilus) microplus and Phlebotomus papatasi: Gene identification, expression and biochemical properties of recombinant proteins

    Science.gov (United States)

    Rhipicephalus (Boophilus) microplus (Bm) is a vector of bovine babesiosis and anaplasmosis. Tick resistance to organophosphate (OP) acaricide involves acetylcholinesterase (AChE) insensitivity to OP and metabolic detoxification. In vitro expression of Bm genes encoding AChE allowed biochemical chara...

  9. Docking and molecular dynamics studies of peripheral site ligand–oximes as reactivators of sarin-inhibited human acetylcholinesterase

    NARCIS (Netherlands)

    Almeida, J.S.F.D. de; Cuya Guizado, T.R.; Guimarães, A.P.; Ramalho, T.C.; Gonçalves, A.S.; Koning, M.C. de; França, T.C.C.

    2016-01-01

    In the present work, we performed docking and molecular dynamics simulations studies on two groups of long-tailored oximes designed as peripheral site binders of acetylcholinesterase (AChE) and potential penetrators on the blood brain barrier. Our studies permitted to determine how the tails anchor

  10. Design and synthesis of dual inhibitors of acetylcholinesterase and serotonin transporter targeting potential agents for Alzheimer's disease.

    Science.gov (United States)

    Kogen, Hiroshi; Toda, Narihiro; Tago, Keiko; Marumoto, Shinji; Takami, Kazuko; Ori, Mayuko; Yamada, Naho; Koyama, Kazuo; Naruto, Shunji; Abe, Kazumi; Yamazaki, Reina; Hara, Takao; Aoyagi, Atsushi; Abe, Yasuyuki; Kaneko, Tsugio

    2002-10-03

    Highly efficient acetylcholinesterase (AChE) and serotonin transporter (SERT) dual inhibitors, (S)-4 and (R)-13 were designed and synthesized on the basis of the hypothetical model of AChE active site. Both compounds showed potent inhibitory activities against AChE and SERT. [structure: see text

  11. Use of cytectrene marked by the technetium 99 to study the activity of Acetylcholinesterase in the rat brain

    International Nuclear Information System (INIS)

    Mejri, Najoua

    2007-01-01

    Alzheimer's disease is a degenerative neurological disorder that causes progressive and irreversible loss of mental functions. It is the most common form of dementia and is characterized by a decrease in serotonergic neurons that carry the 5HT1A receptors. Derivatives piperidine with a tertiary amine and ester are similar to acetylcholine [natural substrate of acetylcholinesterase)], we used the cytectrene [molecule based piperidine marked the technetium 99m] as a substrate to investigate the activity of Acetylcholinesterase in the brain. The use of cytectrene for the quantitative measurement of the activity of the Acetylcholinesterase in the brain depends on the rate of hydrolysis and the enzymatic specificity. The results showed that the cytectrene can be used as a substrate for a precise and quantitative determination of the activity of this enzyme. The use of cytectrene as a substrate of Acetylcholinesterase and determination of its activity can use this molecule as an agent for early diagnosis of Alzheimer's disease. The results will, therefore, not only their importance on a fundamental level but also on a plan applied in the medical field. (Author)

  12. Inhibitory and enzyme-kinetic investigation of chelerythrine and lupeol isolated from Zanthoxylum rhoifolium against krait snake venom acetylcholinesterase

    Energy Technology Data Exchange (ETDEWEB)

    Ahmad, Mustaq, E-mail: mushtaq213@yahoo.com [University of Science and Technology, Bannu, (Pakistan). Department of Biotechnology; Weber, Andrea D.; Zanon, Graciane; Tavares, Luciana de C.; Ilha, Vinicius; Dalcol, Ionara I.; Morel, Ademir F., E-mail: ademirfariasm@gmail.com [Universidade Federal de Santa Maria, RS (Brazil). Dept. de Quimica

    2014-01-15

    The in vitro activity of chelerythrine and lupeol, two metabolites isolated from Zanthoxylum rhoifolium were studied against the venom of the snake Bungarus sindanus (Elapidae). The venom, which is highly toxic to humans, consists mainly by the enzyme acetylcholinesterase (AChE). Both compounds showed activity against the venom, and the alkaloid chelerythrine presented higher activity than did triterpene lupeol. (author)

  13. Development of a bifunctional sensor using haptenized acetylcholinesterase and application for the detection of cocaine and organophosphates

    NARCIS (Netherlands)

    Teller, Carsten; Halamek, J.; Žeravík, Jiri; Stöcklein, Walter F.M.; Scheller, Frieder W.

    2008-01-01

    We developed a dual piezoelectric/amperometric sensor for the detection of two unrelated analytes in one experiment that uses propidium to anchor acetylcholinesterases (AChE) at the surface. This mass-sensitive sensor does not only allow the examination of the interaction between AChE and the

  14. Antidotal effects of varthemia persica DC extract in organophosphate poisoning or warfare agents by measuring whole blood acetylcholinesterase

    International Nuclear Information System (INIS)

    Kalantari, H.; Siahapoosh, A.; Farsani, K. M.

    2009-01-01

    The organophosphates (ORPs) or war fare agents toxicity results from inhibition of acetylcholinesterase (AchE). phosphylation of the active serin of AchE leads to accumulation of acetylcholine in synaptic clefts leading to generalized cholinergic over-stimulation. Standard treatment of ORP poisoning includes a muscarinic antagonist such as Atropine, and acetylcholinesterase reactivator (oxime). Presently, oximes like abidoxime and pralidoxime are approved as antidotes against ORP poisoning but are considered to be rather ineffective against certain ORP. Like Soman. In this study, the protective effect of Varthemia persica DC extract on acetylcholinesterase was examined in rats. Animals in weight range of 200-225 g were divided in 8 groups. The negative control group received only 0.4 ml normal saline, reference group, received ethylparaoxone in dose of 50 percent of LD50, positive control group, received ethylparaoxone (50% LD50) and one minute later 50 mol of pralidoxime. Test group 1: received ethylparaoxone and one minute later single dose of methanolic extract of Varthemia persica (250 mg/kg), Test Group 2: daily received methanolic extract of V.persica (250 mg/kg) in six days and one minute after last dose of extract, ethylparaoxone (50% LD50) were injected, Test Group 3: received ethylparaoxone (50% LD50) and then six doses of methanolic extract of V.persica (250 mg/kg) in six continuous days. Test Group 4: received ethylparaoxone and then single dose of dichloromethane extract of V.persica (250 mg/kg). Test Group 5: received ethylparaoxone and one minute later single high dose of methanolic extract of V.persica (1000 mg/kg). Then blood withdrawn and acetylcholinesterase activity was measured according to modified Ellman's method. Only in groups which received extract of V. persica before and after injection of ethylparaoxone, the mean of acetylcholinesterase activity was significantly different with reference group (p 0.05) but no significant difference with

  15. Increase of acceptability period of Musca domestic l., 1758 (diptera: muscidae) pupae, irradiated by gamma radiation as host of the pupal parasitoid Spalangia endius, Walker, 1839 (hymenoptera: pteromalidae)

    Energy Technology Data Exchange (ETDEWEB)

    Itepan, Natanael M., E-mail: nmitepan@ifsp.edu.br [Instituto Federal de Sao Paulo (IFSP), Piracicaba, SP (Brazil); Itepan, Sara E.D.Z., E-mail: sarazenitepan@ig.com.br [Universidade de Sao Paulo (FFCLRP/USP), Ribeirao Preto, SP (Brazil). Faculdade de Filosofia Ciencias e Letras; Arthur, Valter, E-mail: arthur@cena.usp.br [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil)

    2013-07-01

    This experiment was carried out in Biological Control of Domestic Fly 'Eduardo Hiroshi Mizumoto' Laboratory at Entomology and Acarology Department (LEA/ESALQ/USP) and in Food Irradiation and Radioentomology Laboratory (LIARE/CENA/USP). The gamma radiation source used was a Co-60 irradiator model Gammabeam-650 of the Atomic Energy of Canada Ltd. whose activity in the beginning of the experiments was 9.8x10{sup 13} Bq. (2,644 Ci). The lots of pupae of Musca domestic L., 1758 and the parasitoid Spalangia endius Walker, 1839 had been kept in acclimatized room with 25 ± 2 deg C of temperature and 70 ± 5% of relative humidity. This experiment was designed to investigate if the non-viabilization M. domestic pupae, using gamma radiation, could build up the acceptability period as host of the pupal parasitoid S. endius. At these age intervals, the dose to prevent adult emergence was 25, 220, 360 and 520 Gy respectively. The 1, 2, 3 and 4 days old pupae were irradiated (dose rate: 1,510 Gy/hr) and exposed to the parasitoid S. endius at a proportion of one female parasitoid to five housefly pupae, during different periods after the irradiation. The results allow us to conclude that irradiation increase the acceptability period of the housefly pupae by the parasitoid. The best age to irradiate the housefly pupae was one day. (author)

  16. Increase of acceptability period of Musca domestic l., 1758 (diptera: muscidae) pupae, irradiated by gamma radiation as host of the pupal parasitoid Spalangia endius, Walker, 1839 (hymenoptera: pteromalidae)

    International Nuclear Information System (INIS)

    Itepan, Natanael M.; Itepan, Sara E.D.Z.

    2013-01-01

    This experiment was carried out in Biological Control of Domestic Fly 'Eduardo Hiroshi Mizumoto' Laboratory at Entomology and Acarology Department (LEA/ESALQ/USP) and in Food Irradiation and Radioentomology Laboratory (LIARE/CENA/USP). The gamma radiation source used was a Co-60 irradiator model Gammabeam-650 of the Atomic Energy of Canada Ltd. whose activity in the beginning of the experiments was 9.8x10 13 Bq. (2,644 Ci). The lots of pupae of Musca domestic L., 1758 and the parasitoid Spalangia endius Walker, 1839 had been kept in acclimatized room with 25 ± 2 deg C of temperature and 70 ± 5% of relative humidity. This experiment was designed to investigate if the non-viabilization M. domestic pupae, using gamma radiation, could build up the acceptability period as host of the pupal parasitoid S. endius. At these age intervals, the dose to prevent adult emergence was 25, 220, 360 and 520 Gy respectively. The 1, 2, 3 and 4 days old pupae were irradiated (dose rate: 1,510 Gy/hr) and exposed to the parasitoid S. endius at a proportion of one female parasitoid to five housefly pupae, during different periods after the irradiation. The results allow us to conclude that irradiation increase the acceptability period of the housefly pupae by the parasitoid. The best age to irradiate the housefly pupae was one day. (author)

  17. Improving the knowledge on Piper betle: targeted metabolite analysis and effect on acetylcholinesterase.

    Science.gov (United States)

    Valentão, Patrícia; Gonçalves, Rui F; Belo, Cristóvão; de Pinho, Paula Guedes; Andrade, Paula B; Ferreres, Federico

    2010-10-01

    Piper betle is a species growing in South East Asia, where its leaves are economically and medicinally important. To screen the highest possible number of volatile and semivolatile components, the leaves were subjected to headspace solid-phase microextraction, hydrodistillation and Soxhlet extraction, prior to analysis by GC/MS. Fifty compounds (identified by comparison with standard compounds or tentatively by National Institute of Standards and Technology database) were determined, 23 being described for the first time in this matrix. An aqueous extract was also analysed, in which only seven compounds were characterized. The organic acids' composition of this extract was determined by HPLC/UV and eight compounds are reported for the first time in P. betle. This extract also displayed acetylcholinesterase inhibitory capacity.

  18. Toxicological effect of herbicides (diuron and bentazon) on snake venom and electric eel acetylcholinesterase.

    Science.gov (United States)

    Ahmed, Mushtaq; Latif, Nadia; Khan, Rehmat Ali; Ahmad, Akhlaq

    2012-08-01

    The toxicological effects of the active ingredients of the herbicides diuron and bentazon on the activity of acetylcholinesterase (AChE) of krait (Bungarus sindanus) venom and electric eel (Electrophorus electricus) were studied. The diuron and entazon caused non-competitive inhibition of AChE from both species. For the venom AChE, the calculated IC50 for diuron and bentazon were found to be 3.25 and 0.14 μM, while for eel AChE, the respective IC50 values were 3.6 and 0.135 μM. In comparison, bentazon was a more potent inhibitor than diuron of AChE from both species. The insecticide lindane did not have any inhibitory effect on AChE activity in either species, even when tested at high concentrations (200-800 μM).

  19. Synthesis, Biological Evaluation and Molecular Modelling of 2′-Hydroxychalcones as Acetylcholinesterase Inhibitors

    Directory of Open Access Journals (Sweden)

    Sri Devi Sukumaran

    2016-07-01

    Full Text Available A series of 2′-hydroxy- and 2′-hydroxy-4′,6′-dimethoxychalcones was synthesised and evaluated as inhibitors of human acetylcholinesterase (AChE. The majority of the compounds were found to show some activity, with the most active compounds having IC50 values of 40–85 µM. Higher activities were generally observed for compounds with methoxy substituents in the A ring and halogen substituents in the B ring. Kinetic studies on the most active compounds showed that they act as mixed-type inhibitors, in agreement with the results of molecular modelling studies, which suggested that they interact with residues in the peripheral anionic site and the gorge region of AChE.

  20. Effect of viologen-phosphorus dendrimers on acetylcholinesterase and butyrylcholinesterase activities.

    Science.gov (United States)

    Ciepluch, Karol; Weber, Monika; Katir, Nadia; Caminade, Anne-Marie; El Kadib, Abdelkrim; Klajnert, Barbara; Majoral, Jean Pierre; Bryszewska, Maria

    2013-03-01

    The inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) is the first step in checking whether new compounds can be considered as drugs for treating neurodegenerative diseases. The effect of viologen-phosphorus dendrimers on AChE and BChE activities was studied. The results show that the effects on the cholinesterase activities depend on dendrimer type and size. Viologen dendrimers can interact with the enzymes in two ways: they can bind either to a peripheral site of the enzyme or to amino acids located near the active site, inhibiting catalysis by both cholinesterases. All tested non-toxic viologen-phosphorus dendrimers inhibited the activities of both cholinesterases, showing their potential as new drugs for treating neurodegenerative diseases. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Molecular interaction of acetylcholinesterase with carnosic acid derivatives: a neuroinformatics study.

    Science.gov (United States)

    Merad, M; Soufi, W; Ghalem, S; Boukli, F; Baig, M H; Ahmad, K; Kamal, Mohammad A

    2014-04-01

    Alzheimer's disease is a progressive degenerative disease of the brain marked by gradual and irreversible declines in cognitive functions. Acetylcholinesterase (AChE) plays a biological role in the termination of nerve impulse transmissions at cholinergic synapses by rapid hydrolysis of its substrate, "acetylcholine". The deficit level of acetylcholine leads to deprived nerve impulse transmission. Thus the cholinesterase inhibitors would reverse the deficit in acetylcholine level and consequently may reverse the memory impairments, which is characteristic of the Alzheimer's disease. The molecular interactions between AChE and Carnosic acid, a well known antioxidant substance found in the leaves of the rosemary plant has always been an area of interest. Here in this study we have performed in silico approach to identify carnosic acid derivatives having the potential of being a possible drug candidate against AChE. The best candidates were selected on the basis of the results of different scoring functions.

  2. Detection of the acetylcholinesterase insecticide resistance mutation (G328A) in natural populations of Ceratitis capitata

    International Nuclear Information System (INIS)

    Elfekih, Samia; Haran, Julien; Shannon, Matthew; Vogler, Alfried P.

    2015-01-01

    Wild Mediterranean fruit fly specimens collected from various regions worldwide were screened for the glycine to alanine (Gly->Ala) point mutation (G328A) in the acetylcholinesterase enzyme, presumably causing resistance to organophosphates. We found that the single nucleotide polymorphism (SNP) responsible for this amino acid change is located at the beginning of exon 6 of the Ccace2 gene. The identification of the exact location of the SNP permitted PCR primer design around this site and direct sequencing of the corresponding genomic region. We detected the resistance allele in natural Mediterranean fruit fly populations from Brazil and Spain, but not from other sites in four continents. The known treatment history of sites suggests that the resistance build up is linked to organophosphate application in the held. The PCR-based detection provides a screening method useful for monitoring Mediterranean fruit fly insecticide resistance in local populations and improving pest management strategies accordingly. (author)

  3. Dementia post-radiotherapy: improvement with acetylcholinesterase inhibitor A case report

    Directory of Open Access Journals (Sweden)

    Antonio Eduardo Damin

    Full Text Available Abstract Cognitive decline associated with radiotherapy is a progressive complication that affects many patients submitted to this form of treatment. The lack of an effective treatment drives research for new treatment options to improve the quality of life of patients with this disorder. We report the case of a 64 year-old man who developed a severe dementia of the frontal subcortical type, which was associated with subcortical frontal lesions and appeared as a late complication of radiotherapy used to treat a pituitary tumor. After many pharmacological attempts to improve his cognitive and behavioral problems, the patient showed a significant improvement in the cognitive, functional and behavioral impairments after treatment with an acetylcholinesterase inhibitor. This report discusses hypotheses for the positive effect of this treatment.

  4. A radiotracer for In vivo studies of acetylcholinesterase: p-[{sup 18}F]fluorodonepezil

    Energy Technology Data Exchange (ETDEWEB)

    Lee, S. Y.; Choi, Y. S.; Choi, Y.; Kim, S. E.; Lee, K. H.; Kim, B. T. [Samsung Medical Center, Seoul (Korea, Republic of); Lee, J. W. [Seoul National Univ., Seoul (Korea, Republic of)

    1999-05-01

    Alzheimer's disease (AD) is one of senile dementia caused by lack of acetylcholine in central nervous system, and in vivo studies of acetylcholinesterase (AChE) have been carried out using many radiolabeled AChE inhibitors (donepezil, tacrine, physostigmine, CP-126,998, etc). Donepezil, a FDA approved drug for AD is now in clinical use. Therefore, we synthesized and evaluated p-[{sup 18}F]fluorodonepezil in mice. Biodistribution studies demonstrated that p-[{sup 18}F]fluorodonepezil binds non-specifically in vivo and does not suffer from metabolism in mouse brain. This study suggests that radioligands with higher binding affinity may be required to visualize AChE in vivo and further studies are needed to develop better radiotracers.

  5. Effects of endosulfan on brain acetylcholinesterase activity in juvenile bluegill sunfish

    International Nuclear Information System (INIS)

    Dutta, Hiran M.; Arends, Dane A.

    2003-01-01

    The effects of endosulfan upon brain acetylcholinesterase (AChE) activity were measured in juvenile blue gill sunfish (Lepomis macrochirus). Based on exposure durations of 0, 24, 48, 72, and 96 h and 1 week at 1.0 μg/L (just below the LC50 of 1.2 μg/L for this species), step-wise decreases in AChE activity were noted, corresponding to 0%, 3.57%, 12.65%, 14.23%, 16.31%, and 3.11% inhibition, respectively. Total brain protein concentrations were measured to test the accuracy of the Ache data with no significant anomalies. The duration of exposure was related to the reduction in the AChE activities which reflected the biotoxicity of endosulfan. The changes in the AChE activities will certainly affect the normal behavior of the juvenile blue gill which is detrimental to their very existence in the natural habitat

  6. Detection of Carbofuran with Immobilized Acetylcholinesterase Based on Carbon Nano tubes-Chitosan Modified Electrode

    International Nuclear Information System (INIS)

    Zhang, Sh.; Li, Sh.; Ma, J.; Xiong, F.; Qu, S.; Zhang, Sh.; Li, Sh.

    2013-01-01

    A sensitive and stable enzyme biosensor based on efficient immobilization of acetylcholinesterase (AChE) to MWNTs-modified glassy carbon electrode (GCE) with chitosan (CS) by layer-by-layer (LBL) technique for rapid determination of carbofuran has been devised. According to the inhibitory effect of carbamate pesticide on the enzymatic activity of AChE, we use carbofuran as a model pesticide. The inhibitory effect of carbofuran on the biosensor was proportional to concentration of carbofuran in the range from 10 -10  g/L to 10 -3  g/L with a detection limit of 10 -12  g/L. This biosensor is a promising new method for pesticide analysis

  7. Catalytic recruitment in the inactivation of acetylcholinesterase by soman: temperature dependence of the solvent isotope effect

    International Nuclear Information System (INIS)

    Kovach, I.M.; Huber, J.H.A.; Schowen, R.L.

    1988-01-01

    3,3-Dimethyl-2-butyl methylphosphonofluoridate (soman) recruits at least 60-70% of the catalytic power of acetylcholinesterase during phosphonylation of the active-site serine, as compared with the acetylation by acetylcholine. The solvent isotope effect is 1.34 +/- 0.11 and is temperature independent within experimental error between 5 and 45 0 C. This observation is quite similar to what has been found for the reactions of aryl acetates and anilides. Average activation parameters for the same temperature range are ΔH* = 7.3 +/- 0.6 kcal/mol and ΔS* = -4 +/- 2 eu (H 2 O) and ΔH* = 7.1 +/- 0.4 kcal/mol and ΔS* = -6 +/- 1 eu (D 2 O). A rate-limiting process with 50% contribution of an induced-fit conformational change is supported by the data

  8. Acetylcholinesterase Inhibitory Activities of Flavonoids from the Leaves of Ginkgo biloba against Brown Planthopper

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    Xiao Ding

    2013-01-01

    Full Text Available Ginkgo biloba is a traditional Chinese medicinal plant which has potent insecticidal activity against brown planthopper. The MeOH extract was tested in the acetylcholinesterase (AChE inhibitory assay with IC50 values of 252.1 μg/mL. Two ginkgolides and thirteen flavonoids were isolated from the leaves of Ginkgo biloba. Their structures were established on the basis of spectroscopic data interpretation. It revealed that the 13 isolated flavonoids were found to inhibit AChE with IC50 values ranging from 57.8 to 133.1 μg/mL in the inhibitory assay. AChE was inhibited dose dependently by all tested flavonoids, and compound 6 displayed the highest inhibitory effect against AChE with IC50 values of 57.8 μg/mL.

  9. Synthesis and study of thiocarbonate derivatives of choline as potential inhibitors of acetylcholinesterase.

    Science.gov (United States)

    Boyle, N A; Talesa, V; Giovannini, E; Rosi, G; Norton, S J

    1997-09-12

    Fourteen alkyl and aryl thiocarbonate derivatives of choline were synthesized and studied as potential inhibitors of acetylcholinesterase (AChE). Twelve of the compounds inhibited AChEs derived from calf forebrain, human red blood cells, and octopus brain ranging from low to moderately high inhibition potency. The concentration of each inhibitory compound giving 50% inhibition of enzyme activity (IC50 values, which ranged from 1 x 10(-2) to 8 x 10(-7) M) was determined and is reported; inhibitor constants (Ki values) for the most inhibitory compounds, (1-pentylthiocarbonyl)choline chloride and (1-heptylthiocarbonyl)choline chloride, were calculated from kinetic data and are also reported. The inhibitors are competitive with substrate, and they are not hydrolyzed by the AChE activities. Certain of these new compounds may provide direction for the development of new drugs that have anticholinesterase activity and may be used for the treatment of Alzheimer's disease.

  10. Acetylcholinesterase inhibition-based biosensor for aluminum(III) chronoamperometric determination in aqueous media.

    Science.gov (United States)

    Barquero-Quirós, Miriam; Domínguez-Renedo, Olga; Alonso-Lomillo, Maria Asunción; Arcos-Martínez, María Julia

    2014-05-07

    A novel amperometric biosensor for the determination of Al(III) based on the inhibition of the enzyme acetylcholinesterase has been developed. The immobilization of the enzyme was performed on screen-printed carbon electrodes modified with gold nanoparticles. The oxidation signal of acetylthiocholine iodide enzyme substrate was affected by the presence of Al(III) ions leading to a decrease in the amperometric current. The developed system has a detection limit of 2.1 ± 0.1 μM for Al(III). The reproducibility of the method is 8.1% (n = 4). Main interferences include Mo(VI), W(VI) and Hg(II) ions. The developed method was successfully applied to the determination of Al(III) in spiked tap water . The analysis of a certified standard reference material was also carried out. Both results agree with the certified values considering the respective associated uncertainties.

  11. Surface modification of chitosan/PEO nanofibers by air dielectric barrier discharge plasma for acetylcholinesterase immobilization

    Energy Technology Data Exchange (ETDEWEB)

    Dorraki, Naghme, E-mail: n.dorraki@web.sbu.ac.ir [Laser and Plasma Research Institute, Shahid Beheshti University, Evin 1983963113, Tehran (Iran, Islamic Republic of); Safa, Nasrin Navab [Laser and Plasma Research Institute, Shahid Beheshti University, Evin 1983963113, Tehran (Iran, Islamic Republic of); Jahanfar, Mehdi [Protein Research Center, Shahid Beheshti University, Evin 1983963113, Tehran (Iran, Islamic Republic of); Ghomi, Hamid [Laser and Plasma Research Institute, Shahid Beheshti University, Evin 1983963113, Tehran (Iran, Islamic Republic of); Ranaei-Siadat, Seyed-Omid [Protein Research Center, Shahid Beheshti University, Evin 1983963113, Tehran (Iran, Islamic Republic of)

    2015-09-15

    Highlights: • We used an economical and effective method for surface modification. • Chitosan/PEO nanofibrous membranes were modified by air-DBD plasma. • The most NH{sub 3}{sup +} group was generated on the 6 min plasma modified membrane. • We immobilized acetylcholinesterase on the plasma modified and unmodified membranes. • More enzyme activity was detected on the modified membrane by plasma. - Abstract: There are different methods to modify polymer surfaces for biological applications. In this work we have introduced air-dielectric barrier discharge (DBD) plasma at atmospheric pressure as an economical and safe method for modifying the surface of electrospun chitosan/PEO (90/10) nanofibers for acetylcholinesterase (AChE) immobilization. According to the contact angle measurement results, the nanofibers become highly hydrophilic when they are exposed to the DBD plasma for 6 min in compared to unmodified membrane. Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) results reveal hydroxyl, C=O and NH{sub 3}{sup +} polar groups increment after 6 min plasma treatment. Contact angle measurements and ATR-FTIR results are confirmed by X-ray photoelectron spectroscopy (XPS). AChE at pH 7.4 carries a negative charge and after immobilization on the surface of plasma-treated nanofibrous membrane attracts the NH{sub 3}{sup +} group and more enzyme activity is detected on the plasma-modified nanofibers for 6 min in compared to unmodified nanofibers. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) are used for the surface topography and morphology characterization. The results have proved that air-DBD plasma is a suitable method for chitosan/PEO nanofibrous membrane modification as a biodegradable and functionalized substrate for enzyme immobilization.

  12. Phytochemical Quantification and the In Vitro Acetylcholinesterase Inhibitory Activity of Phellodendron chinense and Its Components.

    Science.gov (United States)

    Kim, Yu Jin; Lim, Hye-Sun; Kim, Yoonju; Lee, Jun; Kim, Bu-Yeo; Jeong, Soo-Jin

    2017-06-02

    The dried bark of Phellodendron chinense has been used as a traditional herbal medicine to remove damp heat, relieve consumptive fever, and cure dysentery and diarrhea. In the present study, we performed quantitative analyses of the two components of P. chinense , phellodendrine and berberine, using high-performance liquid chromatography. A 70% ethanol extract of P. chinense was prepared and the two components were separated on a C-18 analytical column using a gradient solvent system of acetonitrile and 0.1% ( v / v ) aqueous trifluoroacetic acid. The ultraviolet wavelength used for detection was 200 nm for phellodendrine and 226 nm for berberine. The analytical method established here showed high linearity (correlation coefficient, ≥0.9991). The amount of phellodendrine and berberine used was 22.255 ± 0.123 mg/g and 269.651 ± 1.257 mg/g, respectively. Moreover, we performed an in vitro acetylcholinesterase (AChE) activity assay and an amyloid-β aggregation test to examine the biological properties of phellodendrine and berberine as therapeutic drugs for Alzheimer's disease. Phellodendrine and berberine inhibited AChE activity in a dose-dependent manner (IC 50 = 36.51 and 0.44 μM, respectively). In contrast, neither phellodendrine nor berberine had an effect on amyloid-β aggregation. The P. chinense extract and phellodendrine, but not berberine, exhibited antioxidant activity by increasing radical scavenging activity. Moreover, P. chinense demonstrated a neuroprotective effect in hydrogen peroxide-treated HT22 hippocampal cells. Overall, our findings suggest that P. chinense has potential as an anti-Alzheimer's agent via the suppression of the enzymatic activity of acetylcholinesterase and the stimulation of antioxidant activity.

  13. Isoflurane-induced spatial memory impairment in mice is prevented by the acetylcholinesterase inhibitor donepezil.

    Directory of Open Access Journals (Sweden)

    Diansan Su

    Full Text Available Although many studies have shown that isoflurane exposure impairs spatial memory in aged animals, there are no clinical treatments available to prevent this memory deficit. The anticholinergic properties of volatile anesthetics are a biologically plausible cause of cognitive dysfunction in elderly subjects. We hypothesized that pretreatment with the acetylcholinesterase inhibitor donepezil, which has been approved by the Food and Drug Administration (FDA for the treatment of Alzheimer's disease, prevents isoflurane-induced spatial memory impairment in aged mice. In present study, eighteen-month-old mice were administered donepezil (5 mg/kg or an equal volume of saline by oral gavage with a feeding needle for four weeks. Then the mice were exposed to isoflurane (1.2% for six hours. Two weeks later, mice were subjected to the Morris water maze to examine the impairment of spatial memory after exposure to isoflurane. After the behavioral test, the mice were sacrificed, and the protein expression level of acetylcholinesterase (AChE, choline acetylase (ChAT and α7 nicotinic receptor (α7-nAChR were measured in the brain. Each group consisted of 12 mice. We found that isoflurane exposure for six hours impaired the spatial memory of the mice. Compared with the control group, isoflurane exposure dramatically decreased the protein level of ChAT, but not AChE or α7-nAChR. Donepezil prevented isoflurane-induced spatial memory impairments and increased ChAT levels, which were downregulated by isoflurane. In conclusions, pretreatment with the AChE inhibitor donepezil prevented isoflurane-induced spatial memory impairment in aged mice. The mechanism was associated with the upregulation of ChAT, which was decreased by isoflurane.

  14. Molecular interaction studies of acetylcholinesterase with potential acetylcholinesterase inhibitors from the root of Rhodiola crenulata using molecular docking and isothermal titration calorimetry methods.

    Science.gov (United States)

    Li, Fa-Jie; Liu, Yuan; Yuan, Yuan; Yang, Bin; Liu, Zhen-Ming; Huang, Lu-Qi

    2017-11-01

    (-)-Epicatechin gallate ((-)-ECG), 1,2,3,4,6-O-pentagalloylglucose (PGG), rhodionin, herbacetin and rhodiosin isolated from the root of Rhodiola crenulata exhibited potent, dose-dependent inhibitory effects on acetylcholinesterase (AChE) with IC 50 ranged from 57.50±5.83 to 2.43±0.34μg/mL. With the aim of explaining the differences in activity of these active ingredients and clarifying how they inhibit AChE, the AChE-inhibitor interactions were further explored using molecular docking and isothermal titration calorimetry (ITC) methods in the present study. Molecular docking studies revealed that all compounds except PGG showed binding energy values ranging from -10.30 to -8.00kcal/mol while the binding energy of galantamine, a known AChE inhibitor, was -9.53kcal/mol; they inhibited the AChE by binding into the ligand pocket with the similar binding pattern to that of galantamine by interacting with Glu199 of AChE. Inhibition constant of these active ingredients had a positive correlation with binding energy. The interaction between AChE and PGG was further evaluated with the ITC method and the results indicated that the PGG-AChE interaction was relevant to AChE concentration. The results revealed a possible mechanism for the AChE inhibition activity of these bioactive ingredients, which may provide some help in lead compounds optimization in the future. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Two QTL characterized for soft scald and soggy breakdown in apple (Malus × domestica) through pedigree-based analysis of a large population of interconnected families

    NARCIS (Netherlands)

    Howard, Nicholas P.; Weg, van de Eric; Tillman, John; Tong, Cindy B.S.; Silverstein, Kevin A.T.; Luby, James J.

    2018-01-01

    Soft scald and soggy breakdown are important postharvest physiological disorders of apple (Malus × domestica). ‘Honeycrisp’ and some of its offspring are particularly susceptible to developing these disorders. The purpose of this study was to identify molecular markers associated with high

  16. Ectopic expression of class 1 KNOX genes induce and adventitious shoot regeneration and alter growth and development of tobacco (Nicotiana tabacum L) and European plum (Prunus domestica L)

    Science.gov (United States)

    Transgenic plants of tobacco (Nicotiana tabacum L) and plum (Prunus domestica L) were produced by transforming with apple class 1 KNOX genes (MdKN1 and MdKN2) or corn KN1 gene. Transgenic tobacco plants were regenerated in vitro from transformed leaf discs cultured in a tissue medium lacking cytoki...

  17. Genome-wide identification and expression analysis of MAPK and MAPKK gene family in Malus domestica.

    Science.gov (United States)

    Zhang, Shizhong; Xu, Ruirui; Luo, Xiaocui; Jiang, Zesheng; Shu, Huairui

    2013-12-01

    MAPK signal transduction modules play crucial roles in regulating many biological processes in plants, which are composed of three classes of hierarchically organized protein kinases, namely MAPKKKs, MAPKKs, and MAPKs. Although genome-wide analysis of this family has been carried out in some species, little is known about MAPK and MAPKK genes in apple (Malus domestica). In this study, a total of 26 putative apple MAPK genes (MdMPKs) and 9 putative apple MAPKK genes (MdMKKs) have been identified and located within the apple genome. Phylogenetic analysis revealed that MdMAPKs and MdMAPKKs could be divided into 4 subfamilies (groups A, B, C and D), respectively. The predicted MdMAPKs and MdMAPKKs were distributed across 13 out of 17 chromosomes with different densities. In addition, analysis of exon-intron junctions and of intron phase inside the predicted coding region of each candidate gene has revealed high levels of conservation within and between phylogenetic groups. According to the microarray and expressed sequence tag (EST) analysis, the different expression patterns indicate that they may play different roles during fruit development and rootstock-scion interaction process. Moreover, MAPK and MAPKK genes were performed expression profile analyses in different tissues (root, stem, leaf, flower and fruit), and all of the selected genes were expressed in at least one of the tissues tested, indicating that the MAPKs and MAPKKs are involved in various aspects of physiological and developmental processes of apple. To our knowledge, this is the first report of a genome-wide analysis of the apple MAPK and MAPKK gene family. This study provides valuable information for understanding the classification and putative functions of the MAPK signal in apple. © 2013.

  18. Fate of egg proteins during the development of Columba livia domestica embryo.

    Science.gov (United States)

    Shbailat, Seba Jamal; Aslan, Ibtisam Omar

    2018-01-01

    The transfer of egg white into the yolk and consumption of yolk proteins by the embryo are largely unexplored in the pigeon Columba livia domestica. Here, we investigated the route of egg white transfer as well as the degradation and uptake of yolk proteins by the pigeon embryo. Initially, we tested the electrophoretic patterns of proteins in different egg compartments throughout development. Then, we used lysozyme as a reference protein to follow the egg white transfer, and we measured its activity using Micrococcus lysodeikticus as a substrate. Moreover, we determined the general protease activity during different developmental stages in the yolk using casein. Finally, we examined the expression of aminopeptidase-N (APN) and oligopeptide transporter PepT1 genes in the yolk sac membrane (YSM) from incubation day 8 until day 17. Several electrophoretic bands of presumptive egg white proteins appeared in different egg compartments. Also, lysozyme activity was detected chronologically in the egg compartments. It appeared on day 12 in the amniotic and intestinal fluids and on day 14 in the yolk. Moreover, protease activity in the yolk increased significantly on day 14 and thereafter. APN expression was largest on day 8 and reduced generally afterward, whereas PepT1 expression peaked between days 13 and 15 but then reduced substantially. Our results suggest that the egg white proteins move through the amnion and intestine into the yolk where they undergo degradation by the activated proteases. Furthermore, the YSM appears to have a role in protein consumption, and this role decreases toward hatch. © 2018 Wiley Periodicals, Inc.

  19. Manipulation of primary sex ratio in birds: lessons from the homing pigeon (Columba livia domestica).

    Science.gov (United States)

    Goerlich-Jansson, Vivian C; Müller, Martina S; Groothuis, Ton G G

    2013-12-01

    Across various animal taxa not only the secondary sex ratio but also the primary sex ratio (at conception) shows significant deviations from the expected equal proportions of sons and daughters. Birds are especially intriguing to study this phenomenon as avian females are the heterogametic sex (ZW); therefore sex determination might be under direct control of the mother. Avian sex ratios vary in relation to environmental or maternal condition, which can also affect the production of maternal steroids that in turn are involved in reproduction and accumulate in the developing follicle before meiosis. As the proximate mechanisms underlying biased primary sex ratio are largely elusive, we explored how, and to what extent, maternal steroid hormones may be involved in affecting primary or secondary sex ratio in clutches of various species of pigeons. First we demonstrated a clear case of seasonal change in sex ratio in first eggs both in the Rock Pigeon (Columba livia) and in a related species, the Wood Pigeon (Columba palumbus), both producing clutches of two eggs. In the Homing Pigeon (Columba livia domestica), domesticated from the Rock Pigeon, testosterone treatment of breeding females induced a clear male bias, while corticosterone induced a female bias in first eggs and we argue that this is in line with sex allocation theory. We next analyzed treatment effects on follicle formation, yolk mass, and yolk hormones, the latter both pre- and post-ovulatory, in order to test a diversity of potential mechanisms related to both primary and secondary sex ratio manipulation. We conclude that maternal plasma hormone levels may affect several pre-ovulatory mechanisms affecting primary sex ratio, whereas egg hormones are probably involved in secondary sex ratio manipulation only.

  20. Genome-wide analysis of the GH3 family in apple (Malus × domestica).

    Science.gov (United States)

    Yuan, Huazhao; Zhao, Kai; Lei, Hengjiu; Shen, Xinjie; Liu, Yun; Liao, Xiong; Li, Tianhong

    2013-05-02

    Auxin plays important roles in hormone crosstalk and the plant's stress response. The auxin-responsive Gretchen Hagen3 (GH3) gene family maintains hormonal homeostasis by conjugating excess indole-3-acetic acid (IAA), salicylic acid (SA), and jasmonic acids (JAs) to amino acids during hormone- and stress-related signaling pathways. With the sequencing of the apple (Malus × domestica) genome completed, it is possible to carry out genomic studies on GH3 genes to indentify candidates with roles in abiotic/biotic stress responses. Malus sieversii Roem., an apple rootstock with strong drought tolerance and the ancestral species of cultivated apple species, was used as the experimental material. Following genome-wide computational and experimental identification of MdGH3 genes, we showed that MdGH3s were differentially expressed in the leaves and roots of M. sieversii and that some of these genes were significantly induced after various phytohormone and abiotic stress treatments. Given the role of GH3 in the negative feedback regulation of free IAA concentration, we examined whether phytohormones and abiotic stresses could alter the endogenous auxin level. By analyzing the GUS activity of DR5::GUS-transformed Arabidopsis seedlings, we showed that ABA, SA, salt, and cold treatments suppressed the auxin response. These findings suggest that other phytohormones and abiotic stress factors might alter endogenous auxin levels. Previous studies showed that GH3 genes regulate hormonal homeostasis. Our study indicated that some GH3 genes were significantly induced in M. sieversii after various phytohormone and abiotic stress treatments, and that ABA, SA, salt, and cold treatments reduce the endogenous level of axuin. Taken together, this study provides evidence that GH3 genes play important roles in the crosstalk between auxin, other phytohormones, and the abiotic stress response by maintaining auxin homeostasis.

  1. Characterisation of microRNAs from apple (Malus domestica 'Royal Gala') vascular tissue and phloem sap.

    Science.gov (United States)

    Varkonyi-Gasic, Erika; Gould, Nick; Sandanayaka, Manoharie; Sutherland, Paul; MacDiarmid, Robin M

    2010-08-04

    Plant microRNAs (miRNAs) are a class of small, non-coding RNAs that play an important role in development and environmental responses. Hundreds of plant miRNAs have been identified to date, mainly from the model species for which there are available genome sequences. The current challenge is to characterise miRNAs from plant species with agricultural and horticultural importance, to aid our understanding of important regulatory mechanisms in crop species and enable improvement of crops and rootstocks. Based on the knowledge that many miRNAs occur in large gene families and are highly conserved among distantly related species, we analysed expression of twenty-one miRNA sequences in different tissues of apple (Malus x domestica 'Royal Gala'). We identified eighteen sequences that are expressed in at least one of the tissues tested. Some, but not all, miRNAs expressed in apple tissues including the phloem tissue were also detected in the phloem sap sample derived from the stylets of woolly apple aphids. Most of the miRNAs detected in apple phloem sap were also abundant in the phloem sap of herbaceous species. Potential targets for apple miRNAs were identified that encode putative proteins shown to be targets of corresponding miRNAs in a number of plant species. Expression patterns of potential targets were analysed and correlated with expression of corresponding miRNAs. This study validated tissue-specific expression of apple miRNAs that target genes responsible for plant growth, development, and stress response. A subset of characterised miRNAs was also present in the apple phloem translocation stream. A comparative analysis of phloem miRNAs in herbaceous species and woody perennials will aid our understanding of non-cell autonomous roles of miRNAs in plants.

  2. Transcription Profiles Reveal Sugar and Hormone Signaling Pathways Mediating Flower Induction in Apple (Malus domestica Borkh.).

    Science.gov (United States)

    Xing, Li-Bo; Zhang, Dong; Li, You-Mei; Shen, Ya-Wen; Zhao, Cai-Ping; Ma, Juan-Juan; An, Na; Han, Ming-Yu

    2015-10-01

    Flower induction in apple (Malus domestica Borkh.) is regulated by complex gene networks that involve multiple signal pathways to ensure flower bud formation in the next year, but the molecular determinants of apple flower induction are still unknown. In this research, transcriptomic profiles from differentiating buds allowed us to identify genes potentially involved in signaling pathways that mediate the regulatory mechanisms of flower induction. A hypothetical model for this regulatory mechanism was obtained by analysis of the available transcriptomic data, suggesting that sugar-, hormone- and flowering-related genes, as well as those involved in cell-cycle induction, participated in the apple flower induction process. Sugar levels and metabolism-related gene expression profiles revealed that sucrose is the initiation signal in flower induction. Complex hormone regulatory networks involved in cytokinin (CK), abscisic acid (ABA) and gibberellic acid pathways also induce apple flower formation. CK plays a key role in the regulation of cell formation and differentiation, and in affecting flowering-related gene expression levels during these processes. Meanwhile, ABA levels and ABA-related gene expression levels gradually increased, as did those of sugar metabolism-related genes, in developing buds, indicating that ABA signals regulate apple flower induction by participating in the sugar-mediated flowering pathway. Furthermore, changes in sugar and starch deposition levels in buds can be affected by ABA content and the expression of the genes involved in the ABA signaling pathway. Thus, multiple pathways, which are mainly mediated by crosstalk between sugar and hormone signals, regulate the molecular network involved in bud growth and flower induction in apple trees. © The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists.

  3. Facial Mechanosensory Influence on Forelimb Movement in Newborn Opossums, Monodelphis domestica.

    Directory of Open Access Journals (Sweden)

    Marie-Josée Desmarais

    Full Text Available The opossum, Monodelphis domestica, is born very immature but crawls, unaided, with its forelimbs (FL from the mother's birth canal to a nipple where it attaches to pursue its development. What sensory cues guide the newborn to the nipple and trigger its attachment to it? Previous experiments showed that low intensity electrical stimulation of the trigeminal ganglion induces FL movement in in vitro preparations and that trigeminal innervation of the facial skin is well developed in the newborn. The skin does not contain Vater-Pacini or Meissner touch corpuscles at this age, but it contains cells which appear to be Merkel cells (MC. We sought to determine if touch perceived by MC could exert an influence on FL movements. Application of the fluorescent dye AM1-43, which labels sensory cells such as MC, revealed the presence of a large number of labeled cells in the facial epidermis, especially in the snout skin, in newborn opossums. Moreover, calibrated pressure applied to the snout induced bilateral and simultaneous electromyographic responses of the triceps muscle in in vitro preparations of the neuraxis and FL from newborn. These responses increase with stimulation intensity and tend to decrease over time. Removing the facial skin nearly abolished these responses. Metabotropic glutamate 1 receptors being involved in MC neurotransmission, an antagonist of these receptors was applied to the bath, which decreased the EMG responses in a reversible manner. Likewise, bath application of the purinergic type 2 receptors, used by AM1-43 to penetrate sensory cells, also decreased the triceps EMG responses. The combined results support a strong influence of facial mechanosensation on FL movement in newborn opossums, and suggest that this influence could be exerted via MC.

  4. Phytohormone Interaction Modulating Fruit Responses to Photooxidative and Heat Stress on Apple (Malus domestica Borkh.

    Directory of Open Access Journals (Sweden)

    Carolina A. Torres

    2017-12-01

    Full Text Available Sun-related physiological disorders such as sun damage on apples (Malus domestica Borkh are caused by cumulative photooxidative and heat stress during their growing season triggering morphological, physiological, and biochemical changes in fruit tissues not only while it is on the tree but also after it has been harvested. The objective of the work was to establish the interaction of auxin (indole-3-acetic acid; IAA, abscisic acid (ABA, jasmonic acid (JA, salicylic acid (SA, and ethylene (ET and its precursor ACC (free and conjugated, MACC during development of sun-injury-related disorders pre- and post-harvest on apples. Peel tissue was extracted from fruit growing under different sun exposures (Non-exposed, NE; Exposed, EX and with sun injury symptoms (Moderate, Mod. Sampling was carried out every 15 days from 75 days after full bloom (DAFB until 120 days post-harvest in cold storage (1°C, > 90%RH. Concentrations of IAA, ABA, JA, SA, were determined using UHPLC mass spectrometry, and ET and ACC (free and conjugated MACC using gas chromatography. IAA was found not to be related directly to sun injury development, but it decreased 60% in sun exposed tissue, and during fruit development. ABA, JA, SA, and ethylene concentrations were significantly higher (P ≤ 0.05 in Mod tissue, but their concentration, except for ethylene, were not affected by sun exposure. ACC and MACC concentrations increased until 105 DAFB in all sun exposure categories. During post-harvest, ethylene climacteric peak was delayed on EX compared to Mod. ABA and SA concentrations remained stable throughout storage in both tissue. JA dramatically increased post-harvest in both EX and Mod tissue, and orchards, confirming its role in low temperature tolerance. The results suggest that ABA, JA, and SA together with ethylene are modulating some of the abiotic stress defense responses on sun-exposed fruit during photooxidative and heat stress on the tree.

  5. Convergent and divergent evolution of genomic imprinting in the marsupial Monodelphis domestica

    Directory of Open Access Journals (Sweden)

    Das Radhika

    2012-08-01

    Full Text Available Abstract Background Genomic imprinting is an epigenetic phenomenon resulting in parent-of-origin specific monoallelic gene expression. It is postulated to have evolved in placental mammals to modulate intrauterine resource allocation to the offspring. In this study, we determined the imprint status of metatherian orthologues of eutherian imprinted genes. Results L3MBTL and HTR2A were shown to be imprinted in Monodelphis domestica (the gray short-tailed opossum. MEST expressed a monoallelic and a biallelic transcript, as in eutherians. In contrast, IMPACT, COPG2, and PLAGL1 were not imprinted in the opossum. Differentially methylated regions (DMRs involved in regulating imprinting in eutherians were not found at any of the new imprinted loci in the opossum. Interestingly, a novel DMR was identified in intron 11 of the imprinted IGF2R gene, but this was not conserved in eutherians. The promoter regions of the imprinted genes in the opossum were enriched for the activating histone modification H3 Lysine 4 dimethylation. Conclusions The phenomenon of genomic imprinting is conserved in Therians, but the marked difference in the number and location of imprinted genes and DMRs between metatherians and eutherians indicates that imprinting is not fully conserved between the two Therian infra-classes. The identification of a novel DMR at a non-conserved location as well as the first demonstration of histone modifications at imprinted loci in the opossum suggest that genomic imprinting may have evolved in a common ancestor of these two Therian infra-classes with subsequent divergence of regulatory mechanisms in the two lineages.

  6. Beta2-adrenoceptor-mediated tracheal relaxation induced by higenamine from Nandina domestica Thunberg.

    Science.gov (United States)

    Tsukiyama, Muneo; Ueki, Takuro; Yasuda, Yoichi; Kikuchi, Hiroko; Akaishi, Tatsuhiro; Okumura, Hidenobu; Abe, Kazuho

    2009-10-01

    The fruit of Nandina domestica Thunberg (ND, Berberidaceae) has been used to improve cough and breathing difficulties in Japan for many years, but very little is known about the constituent of ND responsible for this effect. We have recently reported that the crude extract from ND (NDE) inhibits histamine- and serotonin-induced contraction of isolated guinea pig trachea, and the inhibitory activity was not explained by nantenine, a well-known alkaloid isolated from ND. To explore other constituent(s) of NDE with tracheal smooth muscle relaxant activity, we fractionated NDE and assessed the pharmacological effects of the fractions using isolated guinea pig tracheal ring preparations. NDE was introduced into a polyaromatic absorbent resin column and stepwise eluted to yield five fractions, among which only the 40 % methanol fraction was active in relaxing tracheal smooth muscle precontracted with histamine. Further separation of the 40 % methanol fraction with high-performance liquid chromatography yielded multiple subfractions, one of which was remarkably active in relaxing histamine-precontracted trachea. Chemical analysis with a time-of-flight mass spectrometer and nuclear magnetic resonance spectrometer identified the constituent of the most active subfraction as higenamine, a benzyltetrahydroisoquinoline alkaloid. The potency and efficacy of the active constituent from NDE in relaxing trachea were almost equivalent to synthetic higenamine. In addition, the effect of the active constituent from NDE was competitively inhibited by the selective beta (2)-adrenoceptor antagonist ICI 118,551. These results indicate that the major constituent responsible for the effect of NDE is higenamine, which probably causes the tracheal relaxation through stimulation of beta (2) adrenoceptors. Georg Thieme Verlag KG Stuttgart-New York.

  7. Detection and molecular status of Isospora sp. from the domestic pigeon (Columba livia domestica).

    Science.gov (United States)

    Matsubara, Ryuma; Fukuda, Yasuhiro; Murakoshi, Fumi; Nomura, Osamu; Suzuki, Toru; Tada, Chika; Nakai, Yutaka

    2017-10-01

    The domestic pigeon, Columba livia domestica, is reared for meat production, as a pet, or for racing. Few reports have characterized the parasitic protists from the genus Isospora isolated from Columbiformes. We detected Isospora-like oocysts from C. livia reared for racing. The oocyst contained two sporocysts, and each sporocyst included four sporozoites. The sporulated oocysts (n=4) were spherical; their mean diameters were 25.6 (24.0-27.2)×24.7 (23.4-26.0) μm. Micropyles, polar granules, and oocyst residuum were absent. The mean length and width of the sporocysts (n=8) were 19.5 (18.5-20.5) and 11.2 (10.2-12.1) μm, respectively. Stieda and sub-Stieda bodies were observed. Single-oocyst PCR revealed two different 18S rRNA gene sequences and one 28S rRNA gene sequence in a single oocyst of Isospora sp. Based on a phylogenetic analysis of the 18S rRNA gene, the two sequences made a group which fell within a cluster of known avian Isospora species. A tree based on the 28S rRNA gene sequence indicated that sequences from the pigeon Isospora sp. fell within a cluster of avian Isospora species. Both trees failed to clarify the phylogenetic relationships among the avian Isospora species due to limited resolution. Because the morphological description of Isospora sp. is based on only four oocysts, Isospora sp. is not proposed as a novel species here. This is the first description of Isospora sp. isolated from the domestic pigeon C. livia. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Amelotin Gene Structure and Expression during Enamel Formation in the Opossum Monodelphis domestica

    Science.gov (United States)

    Gasse, Barbara; Liu, Xi; Corre, Erwan; Sire, Jean-Yves

    2015-01-01

    Amelotin (AMTN) is an ameloblast-secreted protein that belongs to the secretory calcium-binding phosphoprotein family, which also includes the enamel matrix proteins amelogenin, ameloblastin and enamelin. Although AMTN is supposed to play an important role in enamel formation, data were long limited to the rodents, in which it is expressed during the maturation stage. Recent comparative studies in sauropsids and amphibians revealed that (i) AMTN was expressed earlier, i.e. as soon as ameloblasts are depositing the enamel matrix, and (ii) AMTN structure was different, a change which mostly resulted from an intraexonic splicing in the large exon 8 of an ancestral mammal. The present study was performed to know whether the differences in AMTN structure and expression in rodents compared to non-mammalian tetrapods dated back to an early ancestral mammal or were acquired later in mammalian evolution. We sequenced, assembled and screened the jaw transcriptome of a neonate opossum Monodelphis domestica, a marsupial. We found two AMTN transcripts. Variant 1, representing 70.8% of AMTN transcripts, displayed the structure known in rodents, whereas variant 2 (29.2%) exhibited the nonmammalian tetrapod structure. Then, we studied AMTN expression during amelogenesis in a neonate specimen. We obtained similar data as those reported in rodents. These findings indicate that more than 180 million years ago, before the divergence of marsupials and placentals, changes occurred in AMTN function and structure. The spatiotemporal expression was delayed to the maturation stage of amelogenesis and the intraexonic splicing gave rise to isoform 1, encoded by variant 1 and lacking the RGD motif. The ancestral isoform 2, housing the RGD, was initially conserved, as demonstrated here in a marsupial, then secondarily lost in the placental lineages. These findings bring new elements towards our understanding of the non-prismatic to prismatic enamel transition that occurred at the onset of

  9. Amelotin Gene Structure and Expression during Enamel Formation in the Opossum Monodelphis domestica.

    Directory of Open Access Journals (Sweden)

    Barbara Gasse

    Full Text Available Amelotin (AMTN is an ameloblast-secreted protein that belongs to the secretory calcium-binding phosphoprotein family, which also includes the enamel matrix proteins amelogenin, ameloblastin and enamelin. Although AMTN is supposed to play an important role in enamel formation, data were long limited to the rodents, in which it is expressed during the maturation stage. Recent comparative studies in sauropsids and amphibians revealed that (i AMTN was expressed earlier, i.e. as soon as ameloblasts are depositing the enamel matrix, and (ii AMTN structure was different, a change which mostly resulted from an intraexonic splicing in the large exon 8 of an ancestral mammal. The present study was performed to know whether the differences in AMTN structure and expression in rodents compared to non-mammalian tetrapods dated back to an early ancestral mammal or were acquired later in mammalian evolution. We sequenced, assembled and screened the jaw transcriptome of a neonate opossum Monodelphis domestica, a marsupial. We found two AMTN transcripts. Variant 1, representing 70.8% of AMTN transcripts, displayed the structure known in rodents, whereas variant 2 (29.2% exhibited the nonmammalian tetrapod structure. Then, we studied AMTN expression during amelogenesis in a neonate specimen. We obtained similar data as those reported in rodents. These findings indicate that more than 180 million years ago, before the divergence of marsupials and placentals, changes occurred in AMTN function and structure. The spatiotemporal expression was delayed to the maturation stage of amelogenesis and the intraexonic splicing gave rise to isoform 1, encoded by variant 1 and lacking the RGD motif. The ancestral isoform 2, housing the RGD, was initially conserved, as demonstrated here in a marsupial, then secondarily lost in the placental lineages. These findings bring new elements towards our understanding of the non-prismatic to prismatic enamel transition that occurred at

  10. Syringeal specialization of frequency control during song production in the Bengalese finch (Lonchura striata domestica.

    Directory of Open Access Journals (Sweden)

    Kristen R Secora

    Full Text Available Singing in songbirds is a complex, learned behavior which shares many parallels with human speech. The avian vocal organ (syrinx has two potential sound sources, and each sound generator is under unilateral, ipsilateral neural control. Different songbird species vary in their use of bilateral or unilateral phonation (lateralized sound production and rapid switching between left and right sound generation (interhemispheric switching of motor control. Bengalese finches (Lonchura striata domestica have received considerable attention, because they rapidly modify their song in response to manipulations of auditory feedback. However, how the left and right sides of the syrinx contribute to acoustic control of song has not been studied.Three manipulations of lateralized syringeal control of sound production were conducted. First, unilateral syringeal muscular control was eliminated by resection of the left or right tracheosyringeal portion of the hypoglossal nerve, which provides neuromuscular innervation of the syrinx. Spectral and temporal features of song were compared before and after lateralized nerve injury. In a second experiment, either the left or right sound source was devoiced to confirm the role of each sound generator in the control of acoustic phonology. Third, air pressure was recorded before and after unilateral denervation to enable quantification of acoustic change within individual syllables following lateralized nerve resection.These experiments demonstrate that the left sound source produces louder, higher frequency, lower entropy sounds, and the right sound generator produces lower amplitude, lower frequency, higher entropy sounds. The bilateral division of labor is complex and the frequency specialization is the opposite pattern observed in most songbirds. Further, there is evidence for rapid interhemispheric switching during song production. Lateralized control of song production in Bengalese finches may enhance acoustic

  11. Screening of plants used in the European traditional medicine to treat memory disorders for acetylcholinesterase inhibitory activity and anti amyloidogenic activity

    DEFF Research Database (Denmark)

    Lobbens, Eva S B; Vissing, Karina J.; Jorgensen, Lene

    2017-01-01

    and the ability to inhibit the enzymatic activity of acetylcholinesterase was evaluated monitoring the hydrolysis of acetylthiocholine Results Under the experimental conditions investigated, extracts of two plants, Carum carvi and Olea sylvestris, inhibited amyloid beta fibrillation considerably, eight plant...

  12. Characterization of resistance gene analogues (RGAs in apple (Malus × domestica Borkh. and their evolutionary history of the Rosaceae family.

    Directory of Open Access Journals (Sweden)

    Michele Perazzolli

    Full Text Available The family of resistance gene analogues (RGAs with a nucleotide-binding site (NBS domain accounts for the largest number of disease resistance genes and is one of the largest gene families in plants. We have identified 868 RGAs in the genome of the apple (Malus × domestica Borkh. cultivar 'Golden Delicious'. This represents 1.51% of the total number of predicted genes for this cultivar. Several evolutionary features are pronounced in M. domestica, including a high fraction (80% of RGAs occurring in clusters. This suggests frequent tandem duplication and ectopic translocation events. Of the identified RGAs, 56% are located preferentially on six chromosomes (Chr 2, 7, 8, 10, 11, and 15, and 25% are located on Chr 2. TIR-NBS and non-TIR-NBS classes of RGAs are primarily exclusive of different chromosomes, and 99% of non-TIR-NBS RGAs are located on Chr 11. A phylogenetic reconstruction was conducted to study the evolution of RGAs in the Rosaceae family. More than 1400 RGAs were identified in six species based on their NBS domain, and a neighbor-joining analysis was used to reconstruct the phylogenetic relationships among the protein sequences. Specific phylogenetic clades were found for RGAs of Malus, Fragaria, and Rosa, indicating genus-specific evolution of resistance genes. However, strikingly similar RGAs were shared in Malus, Pyrus, and Prunus, indicating high conservation of specific RGAs and suggesting a monophyletic origin of these three genera.

  13. Apple ring rot-responsive putative microRNAs revealed by high-throughput sequencing in Malus × domestica Borkh.

    Science.gov (United States)

    Yu, Xin-Yi; Du, Bei-Bei; Gao, Zhi-Hong; Zhang, Shi-Jie; Tu, Xu-Tong; Chen, Xiao-Yun; Zhang, Zhen; Qu, Shen-Chun

    2014-08-01

    MicroRNAs (miRNAs) are small non-coding RNAs, which silence target mRNA via cleavage or translational inhibition to function in regulating gene expression. MiRNAs act as important regulators of plant development and stress response. For understanding the role of miRNAs responsive to apple ring rot stress, we identified disease-responsive miRNAs using high-throughput sequencing in Malus × domestica Borkh.. Four small RNA libraries were constructed from two control strains in M. domestica, crabapple (CKHu) and Fuji Naga-fu No. 6 (CKFu), and two disease stress strains, crabapple (DSHu) and Fuji Naga-fu No. 6 (DSFu). A total of 59 miRNA families were identified and five miRNAs might be responsive to apple ring rot infection and validated via qRT-PCR. Furthermore, we predicted 76 target genes which were regulated by conserved miRNAs potentially. Our study demonstrated that miRNAs was responsive to apple ring rot infection and may have important implications on apple disease resistance.

  14. The Impact of Different Water Regime on Chlorophyll Fluorescence of Pyrus pyraster L. and Sorbus domestica L

    Directory of Open Access Journals (Sweden)

    Viera Šajbidorová

    2015-01-01

    Full Text Available The water deficit is considered to be significant cause of photosynthesis defects. Measuring of chlorophyll fluorescence is one of the methods revealing defects in the photosynthetic apparatus. The experiment was established with two woody plant (Pyrus pyraster L. and Sorbus domestica L. cultivated in two different regimes of the substrate saturation. The measurement of the modulated fluorescence of chlorophyll a was done by FMS1 fluorometer during three-week period between June and September (2012 and 2013. There were analysed selected parameters of chlorophyll fluorescence: Fv/Fm – maximum quantum efficiency of PSII, ΦPSII – effective quantum yield of PSII and RFD – chlorophyll fluorescence decrease ratio. According to the obtained results, Pyrus pyraster has probably higher potential for adaptation to water deficiency. There were recorded the significant decreases mainly in the values of parameter RFD and ΦPSII for Sorbus domestica within duration of experiment with different water regime in both growing seasons 2012 and 2013. The results document a weak sensitivity of the parameter Fv/Fm on changes in the amount of available water in the substrate in both taxa.

  15. Characterization of Resistance Gene Analogues (RGAs) in Apple (Malus × domestica Borkh.) and Their Evolutionary History of the Rosaceae Family

    Science.gov (United States)

    Baldo, Angela; Righetti, Laura; Bailey, Aubrey; Fontana, Paolo; Velasco, Riccardo; Malnoy, Mickael

    2014-01-01

    The family of resistance gene analogues (RGAs) with a nucleotide-binding site (NBS) domain accounts for the largest number of disease resistance genes and is one of the largest gene families in plants. We have identified 868 RGAs in the genome of the apple (Malus × domestica Borkh.) cultivar ‘Golden Delicious’. This represents 1.51% of the total number of predicted genes for this cultivar. Several evolutionary features are pronounced in M. domestica, including a high fraction (80%) of RGAs occurring in clusters. This suggests frequent tandem duplication and ectopic translocation events. Of the identified RGAs, 56% are located preferentially on six chromosomes (Chr 2, 7, 8, 10, 11, and 15), and 25% are located on Chr 2. TIR-NBS and non-TIR-NBS classes of RGAs are primarily exclusive of different chromosomes, and 99% of non-TIR-NBS RGAs are located on Chr 11. A phylogenetic reconstruction was conducted to study the evolution of RGAs in the Rosaceae family. More than 1400 RGAs were identified in six species based on their NBS domain, and a neighbor-joining analysis was used to reconstruct the phylogenetic relationships among the protein sequences. Specific phylogenetic clades were found for RGAs of Malus, Fragaria, and Rosa, indicating genus-specific evolution of resistance genes. However, strikingly similar RGAs were shared in Malus, Pyrus, and Prunus, indicating high conservation of specific RGAs and suggesting a monophyletic origin of these three genera. PMID:24505246

  16. Characterization of resistance gene analogues (RGAs) in apple (Malus × domestica Borkh.) and their evolutionary history of the Rosaceae family.

    Science.gov (United States)

    Perazzolli, Michele; Malacarne, Giulia; Baldo, Angela; Righetti, Laura; Bailey, Aubrey; Fontana, Paolo; Velasco, Riccardo; Malnoy, Mickael

    2014-01-01

    The family of resistance gene analogues (RGAs) with a nucleotide-binding site (NBS) domain accounts for the largest number of disease resistance genes and is one of the largest gene families in plants. We have identified 868 RGAs in the genome of the apple (Malus × domestica Borkh.) cultivar 'Golden Delicious'. This represents 1.51% of the total number of predicted genes for this cultivar. Several evolutionary features are pronounced in M. domestica, including a high fraction (80%) of RGAs occurring in clusters. This suggests frequent tandem duplication and ectopic translocation events. Of the identified RGAs, 56% are located preferentially on six chromosomes (Chr 2, 7, 8, 10, 11, and 15), and 25% are located on Chr 2. TIR-NBS and non-TIR-NBS classes of RGAs are primarily exclusive of different chromosomes, and 99% of non-TIR-NBS RGAs are located on Chr 11. A phylogenetic reconstruction was conducted to study the evolution of RGAs in the Rosaceae family. More than 1400 RGAs were identified in six species based on their NBS domain, and a neighbor-joining analysis was used to reconstruct the phylogenetic relationships among the protein sequences. Specific phylogenetic clades were found for RGAs of Malus, Fragaria, and Rosa, indicating genus-specific evolution of resistance genes. However, strikingly similar RGAs were shared in Malus, Pyrus, and Prunus, indicating high conservation of specific RGAs and suggesting a monophyletic origin of these three genera.

  17. Development of novel techniques to extract phenolic compounds from Romanian cultivars of Prunus domestica L. and their biological properties.

    Science.gov (United States)

    Mocan, Andrei; Diuzheva, Alina; Carradori, Simone; Andruch, Vasil; Massafra, Chiara; Moldovan, Cadmiel; Sisea, Cristian; Petzer, Jacobus P; Petzer, Anél; Zara, Susi; Marconi, Guya Diletta; Zengin, Gokhan; Crișan, Gianina; Locatelli, Marcello

    2018-04-21

    In the present work, fourteen cultivars of Prunus domestica were analysed to investigate their phenolic pattern with the purpose of using the leaves as potential resources of bioactive compounds in the pharmaceutical and food industry. Microwave-assisted extraction (MAE), dispersive liquid-liquid microextraction and sugaring-out liquid-liquid extraction techniques were optimized in order to obtain an exhaustive multi-component panel of phenolic compounds. The best phenolic-enriched recovery was achieved using MAE in water:methanol (30:70), and this procedure was further applied for quantitative analysis of phenolic compounds in real samples. In order to prove the safeness of these extracts, the biological potential of the Prunus cultivars was tested by several in vitro antioxidant and enzyme inhibitory assays. Moreover, their cytotoxicity was evaluated on human gingival fibroblasts (HGFs), and in most of the cases the treatment with different concentrations of extracts didn't show cytotoxicity up to 500 μg/mL. Only 'Carpatin' and 'Minerva' cultivars, at 250 and 500 μg/mL, reduced partially cell viability of HGFs population. Noteworthy, Centenar cultivar was the most active for the α-glucosidase inhibition (6.77 mmolACAE/g extract), whereas Ialomița cultivar showed the best antityrosinase activity (23.07 mgKAE/g extract). Overall, leaves of P. domestica represent a rich alternative source of bioactive compounds. Copyright © 2018. Published by Elsevier Ltd.

  18. Genome-wide Identification and Expression Analysis of Half-size ABCG Genes in Malus × domestica

    Directory of Open Access Journals (Sweden)

    Juanjuan MA

    2018-03-01

    Full Text Available Half-size adenosine triphosphate-binding cassette transporter subgroup G (ABCG genes play crucial roles in regulating the movements of a variety of substrates and have been well studied in several plants. However, half-size ABCGs have not been characterized in detail in apple (Malus × domestica Borkh.. Here, we performed a genome-wide identification and expression analysis of the half-size ABCG gene family in apple. A total of 46 apple half-size ABCGs were identified and divided into six clusters according to the phylogenetic analysis. A gene structural analysis showed that most half-size ABCGs in the same cluster shared a similar exon–intron organization. A gene duplication analysis showed that segmental, tandem and whole-genome duplications could account for the expansion of half-size ABCG transporters in M. domestica. Moreover, a promoter scan, digital expression analysis and RNA-seq revealed that MdABCG21 may be involved in root's cytokinin transport and that ABCG17 may be involved in the lateral bud development of M. spectabilis ‘Bly114’ by mediating cytokinin transport. The data presented here lay the foundation for further investigations into the biological and physiological processes and functions of half-size ABCG genes in apple. Keywords: apple, ABCG gene, duplication, gene expression

  19. Acetylcholinesterase inhibition starting from extracts of Bauhinia variegata L., Bauhinia var. candida (Aiton) Buch.-Ham., and Bauhinia ungulata L.

    Science.gov (United States)

    Santos, Kamilla Monteiro dos; Gonçalves, Priscila Sant'Ana; Paiva, Maria José Nunes de; Lacerda, Guilherme Araújo

    2011-01-01

    A treatment to the Alzheimer's disease consists inhibition of the acetylcholinesterase, which is responsible for the acetylcholine control in the synapses. We have investigated the potential of inhibition of the acetylcholinesterase produced by hexane extracts of leaves, branches, and flowers from three Bauhinia specimens, which is based on the technique of thin layer chromatography and on identifying the organ of the plant that possesses larger concentration of inhibitors. Retention factor analysis shows values of 0.31aA, 0.31aA, and 0.46aB for flowers B. variegata, B. var. candida, and B. ungulata, respectively. The flower extract of B. ungulata is the most suitable for further studies on this inhibition.

  20. In Vitro Ability of Currently Available Oximes to Reactivate Organophosphate Pesticide-Inhibited Human Acetylcholinesterase and Butyrylcholinesterase

    OpenAIRE

    Kamil Musilek; Kamil Kuca; Daniel Jun; Lucie Musilova

    2011-01-01

    We have in vitro tested the ability of common, commercially available, cholinesterase reactivators (pralidoxime, obidoxime, methoxime, trimedoxime and HI-6) to reactivate human acetylcholinesterase (AChE), inhibited by five structurally different organophosphate pesticides and inhibitors (paraoxon, dichlorvos, DFP, leptophos-oxon and methamidophos). We also tested reactivation of human butyrylcholinesterase (BChE) with the aim of finding a potent oxime, suitable to serve as a “pseudocatalytic...

  1. In Search of an Effective in vivo Reactivator for Organophosphorus Nerve Agent-Inhibited Acetylcholinesterase in the Central Nervous System

    Science.gov (United States)

    2012-01-01

    nerve agents, such as sarin (GB), cyclosarin (GF), and VX, are potent inhibitors of the enzyme cholinesterase (ChE). Their toxic effects are due to...three nerve agents. Keywords: acetylcholinesterase; brain; cholinesterase inhibition; cholinesterase reactivation; cyclosarin; diacetylmonoxime...attributed, at least in part, to nuclophilic impedance [Ekstrom et al., 2006a; b; Hoskovcova et al., 2007]. Other AChE- inhibitors , such as soman, become

  2. Flavonoids, Antioxidant Potential, and Acetylcholinesterase Inhibition Activity of the Extracts from the Gametophyte and Archegoniophore of Marchantia polymorpha L.

    Directory of Open Access Journals (Sweden)

    Xin Wang

    2016-03-01

    Full Text Available Marchantia polymorpha L. is a representative bryophyte used as a traditional Chinese medicinal herb for scald and pneumonia. The phytochemicals in M. polymorpha L. are terpenoids and flavonoids, among which especially the flavonoids show significant human health benefits. Many researches on the gametophyte of M. polymorpha L. have been reported. However, as the reproductive organ of M. polymorpha L., the bioactivity and flavonoids profile of the archegoniophore have not been reported, so in this work the flavonoid profiles, antioxidant and acetylcholinesterase inhibition activities of the extracts from the archegoniophore and gametophyte of M. polymorpha L. were compared by radical scavenging assay methods (DPPH, ABTS, O2−, reducing power assay, acetylcholinesterase inhibition assay and LC-MS analysis. The results showed that the total flavonoids content in the archegoniophore was about 10-time higher than that of the gametophyte. Differences between the archegoniophore and gametophyte of M. polymorpha L. were observed by LC-MS analysis. The archegoniophore extracts showed stronger bio-activities than those of the gametophyte. The archegoniophore extract showed a significant acetylcholinesterase inhibition, while the gametophyte extract hardly inhibited it.

  3. Properties of Dense Cores Embedded in Musca Derived from Extinction Maps and {sup 13}CO, C{sup 18}O, and NH{sub 3} Emission Lines

    Energy Technology Data Exchange (ETDEWEB)

    Machaieie, Dinelsa A.; Vilas-Boas, José W.; Wuensche, Carlos A. [Instituto Nacional de Pesquisas Espaciais, Av. dos Astronautas 1.758—Jardim da Granja, CEP-12227-010, São José dos Campos, SP (Brazil); Racca, Germán A. [Universidade do Estado do Rio Grande do Norte, Faculdade de Ciências Exatas e Naturais/Depto. de Física, Campus Universitário Central, Setor II BR 110, KM 48, Rua Prof. António Campos, Costa e Silva 59610-090—Mossoró-RN (Brazil); Myers, Philip C. [Harvard-Smithsonian Center for Astrophysics, 60 Garden Street, Cambridge, MA 02138 (United States); Hickel, Gabriel R., E-mail: dinelsa.machaieie@inpe.br [Universidade Federal de Itajubá, Departamento de Física, Av. BPS, 1303, Pinheirinho, Itajubá, MG (Brazil)

    2017-02-10

    Using near-infrared data from the Two Micron All Sky Survey catalog and the Near Infrared Color Excess method, we studied the extinction distribution in five dense cores of Musca, which show visual extinction greater than 10 mag and are potential sites of star formation. We analyzed the stability in four of them, fitting their radial extinction profiles with Bonnor–Ebert isothermal spheres, and explored their properties using the J = 1–0 transition of {sup 13}CO and C{sup 18}O and the J = K = 1 transition of NH{sub 3}. One core is not well described by the model. The stability parameter of the fitted cores ranges from 4.5 to 5.7 and suggests that all cores are stable, including Mu13, which harbors one young stellar object (YSO), the IRAS 12322-7023 source. However, the analysis of the physical parameters shows that Mu13 tends to have larger A {sub V}, n {sub c}, and P {sub ext} than the remaining starless cores. The other physical parameters do not show any trend. It is possible that those are the main parameters to explore in active star-forming cores. Mu13 also shows the most intense emission of NH{sub 3}. Its {sup 13}CO and C{sup 18}O lines have double peaks, whose integrated intensity maps suggest that they are due to the superposition of clouds with different radial velocities seen in the line of sight. It is not possible to state whether these clouds are colliding and inducing star formation or are related to a physical process associated with the formation of the YSO.

  4. Distribution of intravenously administered acetylcholinesterase inhibitor and acetylcholinesterase activity in the adrenal gland: 11C-donepezil PET study in the normal rat.

    Science.gov (United States)

    Watabe, Tadashi; Naka, Sadahiro; Ikeda, Hayato; Horitsugi, Genki; Kanai, Yasukazu; Isohashi, Kayako; Ishibashi, Mana; Kato, Hiroki; Shimosegawa, Eku; Watabe, Hiroshi; Hatazawa, Jun

    2014-01-01

    Acetylcholinesterase (AChE) inhibitors have been used for patients with Alzheimer's disease. However, its pharmacokinetics in non-target organs other than the brain has not been clarified yet. The purpose of this study was to evaluate the relationship between the whole-body distribution of intravenously administered (11)C-Donepezil (DNP) and the AChE activity in the normal rat, with special focus on the adrenal glands. The distribution of (11)C-DNP was investigated by PET/CT in 6 normal male Wistar rats (8 weeks old, body weight  = 220 ± 8.9 g). A 30-min dynamic scan was started simultaneously with an intravenous bolus injection of (11)C-DNP (45.0 ± 10.7 MBq). The whole-body distribution of the (11)C-DNP PET was evaluated based on the Vt (total distribution volume) by Logan-plot analysis. A fluorometric assay was performed to quantify the AChE activity in homogenized tissue solutions of the major organs. The PET analysis using Vt showed that the adrenal glands had the 2nd highest level of (11)C-DNP in the body (following the liver) (13.33 ± 1.08 and 19.43 ± 1.29 ml/cm(3), respectively), indicating that the distribution of (11)C-DNP was the highest in the adrenal glands, except for that in the excretory organs. The AChE activity was the third highest in the adrenal glands (following the small intestine and the stomach) (24.9 ± 1.6, 83.1 ± 3.0, and 38.5 ± 8.1 mU/mg, respectively), indicating high activity of AChE in the adrenal glands. We demonstrated the whole-body distribution of (11)C-DNP by PET and the AChE activity in the major organs by fluorometric assay in the normal rat. High accumulation of (11)C-DNP was observed in the adrenal glands, which suggested the risk of enhanced cholinergic synaptic transmission by the use of AChE inhibitors.

  5. Distribution of intravenously administered acetylcholinesterase inhibitor and acetylcholinesterase activity in the adrenal gland: 11C-donepezil PET study in the normal rat.

    Directory of Open Access Journals (Sweden)

    Tadashi Watabe

    Full Text Available PURPOSE: Acetylcholinesterase (AChE inhibitors have been used for patients with Alzheimer's disease. However, its pharmacokinetics in non-target organs other than the brain has not been clarified yet. The purpose of this study was to evaluate the relationship between the whole-body distribution of intravenously administered (11C-Donepezil (DNP and the AChE activity in the normal rat, with special focus on the adrenal glands. METHODS: The distribution of (11C-DNP was investigated by PET/CT in 6 normal male Wistar rats (8 weeks old, body weight  = 220 ± 8.9 g. A 30-min dynamic scan was started simultaneously with an intravenous bolus injection of (11C-DNP (45.0 ± 10.7 MBq. The whole-body distribution of the (11C-DNP PET was evaluated based on the Vt (total distribution volume by Logan-plot analysis. A fluorometric assay was performed to quantify the AChE activity in homogenized tissue solutions of the major organs. RESULTS: The PET analysis using Vt showed that the adrenal glands had the 2nd highest level of (11C-DNP in the body (following the liver (13.33 ± 1.08 and 19.43 ± 1.29 ml/cm(3, respectively, indicating that the distribution of (11C-DNP was the highest in the adrenal glands, except for that in the excretory organs. The AChE activity was the third highest in the adrenal glands (following the small intestine and the stomach (24.9 ± 1.6, 83.1 ± 3.0, and 38.5 ± 8.1 mU/mg, respectively, indicating high activity of AChE in the adrenal glands. CONCLUSIONS: We demonstrated the whole-body distribution of (11C-DNP by PET and the AChE activity in the major organs by fluorometric assay in the normal rat. High accumulation of (11C-DNP was observed in the adrenal glands, which suggested the risk of enhanced cholinergic synaptic transmission by the use of AChE inhibitors.

  6. Antimicrobial and acetylcholinesterase inhibitory activities of Buddleja salviifolia (L.) Lam. leaf extracts and isolated compounds.

    Science.gov (United States)

    Pendota, S C; Aderogba, M A; Ndhlala, A R; Van Staden, J

    2013-07-09

    Buddleja salviifolia leaves are used for the treatment of eye infections and neurodegenerative conditions by various tribes in South Africa. This study was designed to isolate the phenolic constituents from the leaf extracts of Buddleja salviifolia and evaluate their antimicrobial and acetylcholinesterase (AChE) activities. Three phenolic compounds were isolated from the ethyl acetate fraction of a 20% aqueous methanol leaf extract of Buddleja salviifolia using Sephadex LH-20 and silica gel columns. Structure elucidation of the isolated compounds was carried out using spectroscopic techniques: mass spectrometry (ESI-TOF-MS) and NMR (1D and 2D). The extracts and isolated compounds were evaluated for antimicrobial and acetylcholinesterase activities using the microdilution technique. The bacteria used for the antimicrobial assays were Gram-positive Bacillus subtilis and Staphylococcus aureus and Gram-negative Escherichia coli and Klebsiella pneumoniae. The isolated compounds were characterized as: 4'-hydroxyphenyl ethyl vanillate (1) a new natural product, acteoside (2) and quercetin (3). The crude extract, fractions and the isolated compounds from the leaves of the plant exhibited a broad spectrum of antibacterial activity. The EtOAc fraction exhibited good activity against Bacillus subtilis and Staphylococcus aureus with MIC values ranging from 780.0 to 390.0 µg/mL. Isolated compound 2 exhibited good activity against Staphylococcus aureus with an MIC value of 62.5 µg/mL. The hexane and DCM fractions of leaves showed the best activity against Candida albicans with MIC and MFC values of 390.0 µg/mL. In the AChE inhibitory test, among the tested extracts, the hexane fraction was the most potent with an IC50 value of 107.4 µg/mL, whereas for the isolated compounds, it was compound (3) (quercetin) with an IC50 value of 66.8 µg/mL. Activities demonstrated by the extracts and isolated compounds support the ethnopharmacological use of Buddleja salviifolia against eye

  7. Sodium-hydrogen exchanger inhibitory potential of Malus domestica, Musa × paradisiaca, Daucus carota, and Symphytum officinale.

    Science.gov (United States)

    Verma, Vivek; Singh, Nirmal; Jaggi, Amteshwar Singh

    2014-02-01

    The involvement of sodium-hydrogen exchangers (NHE) has been described in the pathophysiology of diseases including ischemic heart and brain diseases, cardiomyopathy, congestive heart failure, epilepsy, dementia, and neuropathic pain. Synthetic NHE inhibitors have not achieved much clinical success; therefore, plant-derived phytoconstituents may be explored as NHE inhibitors. In the present study, the NHE inhibitory potential of hydroalcoholic and alkaloidal fractions of Malus domestica, Musa × paradisiaca, Daucus carota, and Symphytum officinale was evaluated. The different concentrations of hydroalcoholic and alkaloidal extracts of the selected plants were evaluated for their NHE inhibitory activity in the platelets using the optical swelling assay. Among the hydroalcoholic extracts, the highest NHE inhibitory activity was shown by M. domestica (IC50=2.350 ± 0.132 μg/mL) followed by Musa × paradisiaca (IC50=7.967 ± 0.451 μg/mL), D. carota (IC50=37.667 ± 2.517 μg/mL), and S. officinale (IC50=249.330 ± 1.155 μg/mL). Among the alkaloidal fractions, the highest NHE inhibitory activity was shown by the alkaloidal fraction of Musa × paradisiacal (IC50=0.010 ± 0.001 μg/mL) followed by D. carota (IC50=0.024 ± 0.002 μg/mL), M. domestica (IC50=0.031 ± 0.005 μg/mL), and S. officinale (IC50=4.233 ± 0.379 μg/mL). The IC50 of alkaloidal fractions was comparable to the IC50 of synthetic NHE inhibitor, EIPA [5-(N-ethyl-N-isopropyl)amiloride] (IC50=0.033 ± 0.004 μg/mL). It may be concluded that the alkaloidal fractions of these plants possess potent NHE inhibitory activity and may be exploited for their therapeutic potential in NHE activation-related pathological complications.

  8. Improving the acetylcholinesterase inhibitory effect of Illigera henryi by solid-state fermentation with Clonostachys rogersoniana.

    Science.gov (United States)

    Li, Xue-Jiao; Dong, Jian-Wei; Cai, Le; Mei, Rui-Feng; Ding, Zhong-Tao

    2017-11-01

    Illigera henryi, an endemic traditional Chinese medicine, contains abundant aporphine alkaloids that possess various bioactivities. In the present study, tubers of I. henryi were fermented by several fungi, and the acetylcholinesterase (AChE) inhibitory activities of non-fermented and fermented I. henryi were measured. The results showed that the fermentation of I. henryi with Clonostachys rogersoniana 828H2 is effective for improving the AChE inhibitory activity. A key biotransformation was found during the C. rogersoniana fermentation for clarifying the improvement of the AChE inhibitory activity of I. henryi: (S)-actinodaphnine (1) was converted to a new 4-hydroxyaporphine alkaloid (4R,6aS)-4-hydroxyactinodaphnine (2) that possessed a stronger AChE inhibitory activity, with an IC 50 value of 17.66±0.06 μM. This paper is the first to report that the pure strain fermentation processing of I. henryi and indicated C. rogersoniana fermentation might be a potential processing method for I. henryi. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  9. Neuromuscular Junction Impairment in Amyotrophic Lateral Sclerosis: Reassessing the Role of Acetylcholinesterase.

    Science.gov (United States)

    Campanari, Maria-Letizia; García-Ayllón, María-Salud; Ciura, Sorana; Sáez-Valero, Javier; Kabashi, Edor

    2016-01-01

    Amyotrophic Lateral Sclerosis (ALS) is a highly debilitating disease caused by progressive degeneration of motorneurons (MNs). Due to the wide variety of genes and mutations identified in ALS, a highly varied etiology could ultimately converge to produce similar clinical symptoms. A major hypothesis in ALS research is the "distal axonopathy" with pathological changes occurring at the neuromuscular junction (NMJ), at very early stages of the disease, prior to MNs degeneration and onset of clinical symptoms. The NMJ is a highly specialized cholinergic synapse, allowing signaling between muscle and nerve necessary for skeletal muscle function. This nerve-muscle contact is characterized by the clustering of the collagen-tailed form of acetylcholinesterase (ColQ-AChE), together with other components of the extracellular matrix (ECM) and specific key molecules in the NMJ formation. Interestingly, in addition to their cholinergic role AChE is thought to play several "non-classical" roles that do not require catalytic function, most prominent among these is the facilitation of neurite growth, NMJ formation and survival. In all this context, abnormalities of AChE content have been found in plasma of ALS patients, in which AChE changes may reflect the neuromuscular disruption. We review these findings and particularly the evidences of changes of AChE at neuromuscular synapse in the pre-symptomatic stages of ALS.

  10. Binding free energy calculations to rationalize the interactions of huprines with acetylcholinesterase.

    Science.gov (United States)

    Nascimento, Érica C M; Oliva, Mónica; Andrés, Juan

    2018-05-01

    In the present study, the binding free energy of a family of huprines with acetylcholinesterase (AChE) is calculated by means of the free energy perturbation method, based on hybrid quantum mechanics and molecular mechanics potentials. Binding free energy calculations and the analysis of the geometrical parameters highlight the importance of the stereochemistry of huprines in AChE inhibition. Binding isotope effects are calculated to unravel the interactions between ligands and the gorge of AChE. New chemical insights are provided to explain and rationalize the experimental results. A good correlation with the experimental data is found for a family of inhibitors with moderate differences in the enzyme affinity. The analysis of the geometrical parameters and interaction energy per residue reveals that Asp72, Glu199, and His440 contribute significantly to the network of interactions between active site residues, which stabilize the inhibitors in the gorge. It seems that a cooperative effect of the residues of the gorge determines the affinity of the enzyme for these inhibitors, where Asp72, Glu199, and His440 make a prominent contribution.

  11. Calcium-activated butyrylcholinesterase in human skin protects acetylcholinesterase against suicide inhibition by neurotoxic organophosphates

    International Nuclear Information System (INIS)

    Schallreuter, Karin U.; University of Bradford; Elwary, Souna M.; Parkin, Susan M.; Wood, John M.

    2007-01-01

    The human epidermis holds an autocrine acetylcholine production and degradation including functioning membrane integrated and cytosolic butyrylcholinesterase (BuchE). Here we show that BuchE activities increase 9-fold in the presence of calcium (0.5 x 10 -3 M) via a specific EF-hand calcium binding site, whereas acetylcholinesterase (AchE) is not affected. 45 Calcium labelling and computer simulation confirmed the presence of one EF-hand binding site per subunit which is disrupted by H 2 O 2 -mediated oxidation. Moreover, we confirmed the faster hydrolysis by calcium-activated BuchE using the neurotoxic organophosphate O-ethyl-O-(4-nitrophenyl)-phenylphosphonothioate (EPN). Considering the large size of the human skin with 1.8 m 2 surface area with its calcium gradient in the 10 -3 M range, our results implicate calcium-activated BuchE as a major protective mechanism against suicide inhibition of AchE by organophosphates in this non-neuronal tissue

  12. Morphometry and acetylcholinesterase activity of the myenteric plexus of the wild mouse Calomys callosus

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    L.B.M. Maifrino

    1997-05-01

    Full Text Available The myenteric plexus of the digestive tract of the wild mouse Calomys callosus was examined using a histochemical method that selectively stains nerve cells, and the acetylcholinesterase (AChE histochemical technique in whole-mount preparations. Neuronal density was 1,500 ± 116 neurons/cm2 (mean ± SEM in the esophagus, 8,900 ± 1,518 in the stomach, 9,000 ± 711 in the jejunum and 13,100 ± 2,089 in the colon. The difference in neuronal density between the esophagus and other regions was statistically significant. The neuron profile area ranged from 45 to 1,100 µm2. The difference in nerve cell size between the jejunum and other regions was statistically significant. AChE-positive nerve fibers were distributed within the myenteric plexus which is formed by a primary meshwork of large nerve bundles and a secondary meshwork of finer nerve bundles. Most of the nerve cells displayed AChE activity in the cytoplasm of different reaction intensities. These results are important in order to understand the changes occurring in the myenteric plexus in experimental Chagas' disease

  13. Chemical Composition and Acetylcholinesterase Inhibitory Activity of Essential Oils from Piper Species.

    Science.gov (United States)

    Xiang, Cai-Peng; Han, Jia-Xin; Li, Xing-Cong; Li, Yun-Hui; Zhang, Yi; Chen, Lin; Qu, Yan; Hao, Chao-Yun; Li, Hai-Zhou; Yang, Chong-Ren; Zhao, San-Jun; Xu, Min

    2017-05-10

    The essential oils (EOs) derived from aromatic plants such as Piper species are considered to play a role in alleviating neuronal ailments that are associated with inhibition of acetylcholinesterase (AChE). The chemical compositions of 23 EOs prepared from 16 Piper spp. were analyzed by both gas chromatography with a flame ionization detector (GC-FID) and gas chromatography-mass spectrometry (GC-MS). A total of 76 compounds were identified in the EOs from the leaves and stems of 19 samples, while 30 compounds were detected in the EOs from the fruits of four samples. Sesquiterpenes and phenylpropanoids were found to be rich in these EOs, of which asaricin, caryophyllene, caryophyllene oxide, isospathulenol, (+)-spathulenol, and β-bisabolene are the major constituents. The EOs from the leaves and stems of Piper austrosinense, P. puberulum, P. flaviflorum, P. betle, and P. hispidimervium showed strong AChE inhibitory activity with IC 50 values in the range of 1.51 to 13.9 mg/mL. A thin-layer chromatography (TLC) bioautography assay was employed to identify active compound(s) in the most active EO from P. hispidimervium. The active compound was isolated and identified as asaricin, which gave an IC 50 value of 0.44 ± 0.02 mg/mL against AChE, comparable to galantamine with an IC 50 0.15 ± 0.01 mg/mL.

  14. Acetylcholinesterase and butyrylcholinesterase inhibitory activity of Pinus species essential oils and their constituents.

    Science.gov (United States)

    Bonesi, Marco; Menichini, Federica; Tundis, Rosa; Loizzo, Monica R; Conforti, Filomena; Passalacqua, Nicodemo G; Statti, Giancarlo A; Menichini, Francesco

    2010-10-01

    This study aimed to investigate the acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activity of the essential oils from Pinus nigra subsp. nigra, P. nigra var. calabrica, and P. heldreichii subsp. leucodermis. This activity is relevant to the treatment of Alzheimer's disease (AD), since cholinesterase drugs are currently the only drugs available to treat AD. P. heldreichii subsp. leucodermis exhibited the most promising activity, with IC(50) values of 51.1 and 80.6 microg/mL against AChE and BChE, respectively. An interesting activity against AChE was also observed with P. nigra subsp. nigra essential oil, with an IC(50) value of 94.4 microg/mL. Essential oils were analyzed by GC and GC-MS with the purpose of investigating their relationships with the observed activities. Among the identified constituents, terpinolene, beta-phellandrene, linalyl acetate, trans-caryophyllene, and terpinen-4-ol were tested. trans-Caryophyllene and terpinen-4-ol inhibited BChE with IC(50) values of 78.6 and 107.6 microg/mL, respectively. beta-Phellandrene was selective against AChE (IC(50) value of 120.2 microg/mL).

  15. Effects of Chlorophenoxy Herbicides and Their Main Transformation Products on DNA Damage and Acetylcholinesterase Activity

    Directory of Open Access Journals (Sweden)

    Sofia Benfeito

    2014-01-01

    Full Text Available Persistent pesticide transformation products (TPs are increasingly being detected among different environmental compartments, including groundwater and surface water. However, there is no sufficient experimental data on their toxicological potential to assess the risk associated with TPs, even if their occurrence is known. In this study, the interaction of chlorophenoxy herbicides (MCPA, mecoprop, 2,4-D and dichlorprop and their main transformation products with calf thymus DNA by UV-visible absorption spectroscopy has been assessed. Additionally, the toxicity of the chlorophenoxy herbicides and TPs was also assessed evaluating the inhibition of acetylcholinesterase activity. On the basis of the results found, it seems that AChE is not the main target of chlorophenoxy herbicides and their TPs. However, the results found showed that the transformation products displayed a higher inhibitory activity when compared with the parent herbicides. The results obtained in the DNA interaction studies showed, in general, a slight effect on the stability of the double helix. However, the data found for 4-chloro-2-methyl-6-nitrophenol suggest that this transformation product can interact with DNA through a noncovalent mode.

  16. Acetylcholinesterase concentrations in heparinized blood of Hispaniolan Amazon parrots (Amazona ventralis).

    Science.gov (United States)

    Tully, Thomas N; Osofsky, Anna; Jowett, Peter L H; Hosgood, Giselle

    2003-12-01

    Organophosphate and carbamate pesticides inhibit acetylcholinesterase (AChE) at nerve synapses. Blood samples from 22 Hispaniolan Amazon parrots (Amazona ventralis) were assayed for cholinesterase levels by two different techniques. Using the modified Michel method, the whole-blood cholinesterase activity levels ranged from 0.082 to 0.616 deltapH/hr with a mean value of 0.35 deltapH/hr. A reference range (0.08-0.62 deltapH/hr) for cholinesterase was established in birds. The modified Ellman spectrophotometric method was used to measure AChE activity by adding acetylthiocholine or pseudocholinesterase (plasma cholinesterase) activity by adding butyrylthiocholine. The reference range of the AChE activity using the modified Ellman spectrophotometric method was 0-1.12 micromol/ml/min with a mean of 0.48 micromol/ml/min, and for pseudocholinesterase the range was 0.09-0.98 micromol/ml/min with a mean of 0.53 micromol/ml/min.

  17. Nanoparticle-based immunosensor with apoferritin templated metallic phosphate label for quantification of phosphorylated acetylcholinesterase

    International Nuclear Information System (INIS)

    Du, Dan; Chen, Aiqiong; Xie, Yunying; Zhang, Aidong; Lin, Yuehe

    2011-01-01

    A new sandwich-like electrochemical immunosensor has been developed for quantification of organophosphorylated acetylcholinesterase (OP-AChE), an exposure biomarker of organophosphate pesticides and nerve agents. Zirconia nanoparticles (ZrO2 NPs) were anchored on a screen printed electrode (SPE) to preferably capture OP-AChE adducts by metal chelation with phospho-moieties, which was selectively recognized by lead phosphate-apoferritin labeled anti-AChE antibody (LPA-anti-AChE). The sandwich-like immunoreactions were performed among ZrO2 NPs, OP-AChE and LPA-anti-AChE to form ZrO2/OP-AChE/LPA-anti-AChE complex and the released lead ions were detected on a disposable SPE. The binding affinity was investigated by both square wave voltammetry (SWV) and quartz crystal microbalance (QCM) measurements. The proposed immunosensor yielded a linear response current over a broad OP-AChE concentrations range from 0.05 nM to 10 nM, with detection limit of 0.02 nM, which has enough sensitivity for monitoring of low-dose exposure to OPs. This method avoids the drawback of unavailability of commercial OP-specific antibody as well as amplifies detection signal by using apoferritin encoded metallic phosphate nanoparticle tags. This nanoparticle-based immunosensor offers a new method for rapid, sensitive, selective and inexpensive quantification of phosphorylated adducts for monitoring of OP pesticides and nerve agents exposures.

  18. Acetylcholinesterase inhibitory activity of Thai traditional nootropic remedy and its herbal ingredients.

    Science.gov (United States)

    Tappayuthpijarn, Pimolvan; Itharat, Arunporn; Makchuchit, Sunita

    2011-12-01

    The incidence of Alzheimer disease (AD) is increasing every year in accordance with the increasing of elderly population and could pose significant health problems in the future. The use of medicinal plants as an alternative prevention or even for a possible treatment of the AD is, therefore, becoming an interesting research issue. Acetylcholinesterase (AChE) inhibitors are well-known drugs commonly used in the treatment of AD. The aim of the present study was to screen for AChE inhibitory activity of the Thai traditional nootropic recipe and its herbal ingredients. The results showed that ethanolic extracts of four out of twenty-five herbs i.e. Stephania pierrei Diels. Kaempfera parviflora Wall. ex Baker, Stephania venosa (Blume) Spreng, Piper nigrum L at 0.1 mg/mL showed % AChE inhibition of 89, 64, 59, 50; the IC50 were 6, 21, 29, 30 microg/mL respectively. The other herbs as well as combination of the whole recipe had no synergistic inhibitory effect on AChE activity. However some plants revealed antioxidant activity. More research should have be performed on this local wisdom remedy to verify the uses in scientific term.

  19. Efficacy of acetylcholinesterase inhibitors versus nootropics in Alzheimer's disease: a retrospective, longitudinal study.

    Science.gov (United States)

    Tsolaki, M; Pantazi, T; Kazis, A

    2001-01-01

    The aim of this study was to investigate the efficacy of nootropics (piracetam, aniracetam, nimodopine and dihydroergicristine) versus acetylcholinesterase inhibitors (AChE-Is) (tacrine and donepezil) in the treatment of Alzheimer's disease. This is a retrospective study of 510 patients with Alzheimer's disease. To determine clinical efficacy of treatment, we used the mean change over time in scores for the following tests: the Mini-Mental State Examination (MMSE); the Cambridge Cognitive Examination for the Elderly; and the Functional Rating Scale for Symptoms of Dementia. In all patients and in patients with severe Alzheimer's disease (baseline MMSE nootropics (-4.38 for AChE-Is group versus 1.48 for nootropics group). For patients with mild dementia (baseline MMSE score between 21 and 26), there was a significantly greater deterioration on the MMSE scale for each time-point in the nootropics group compared with the AChE-Is group. In conclusion, we did not find any strong evidence that a difference in efficacy exists between AChE-Is and nootropics in the treatment of Alzheimer's disease.

  20. New Cinchona Oximes Evaluated as Reactivators of Acetylcholinesterase and Butyrylcholinesterase Inhibited by Organophosphorus Compounds

    Directory of Open Access Journals (Sweden)

    Maja Katalinić

    2017-07-01

    Full Text Available For the last six decades, researchers have been focused on finding efficient reactivators of organophosphorus compound (OP-inhibited acetylcholinesterase (AChE and butyrylcholinesterase (BChE. In this study, we have focused our research on a new oxime scaffold based on the Cinchona structure since it was proven to fit the cholinesterases active site and reversibly inhibit their activity. Three Cinchona oximes (C1, C2, and C3, derivatives of the 9-oxocinchonidine, were synthesized and investigated in reactivation of various OP-inhibited AChE and BChE. As the results showed, the tested oximes were more efficient in the reactivation of BChE and they reactivated enzyme activity to up to 70% with reactivation rates similar to known pyridinium oximes used as antidotes in medical practice today. Furthermore, the oximes showed selectivity towards binding to the BChE active site and the determined enzyme-oxime dissociation constants supported work on the future development of inhibitors in other targeted studies (e.g., in treatment of neurodegenerative disease. Also, we monitored the cytotoxic effect of Cinchona oximes on two cell lines Hep G2 and SH-SY5Y to determine the possible limits for in vivo application. The cytotoxicity results support future studies of these compounds as long as their biological activity is targeted in the lower micromolar range.

  1. Multiple ace genes encoding acetylcholinesterases of Caenorhabditis elegans have distinct tissue expression.

    Science.gov (United States)

    Combes, Didier; Fedon, Yann; Toutant, Jean-Pierre; Arpagaus, Martine

    2003-08-01

    ace-1 and ace-2 genes encoding acetylcholinesterase in the nematode Caenorhabditis elegans present 35% identity in coding sequences but no homology in noncoding regions (introns, 5'- and 3'-untranslated regions). A 5'-region of ace-2 was defined by rescue of ace-1;ace-2 mutants. When green fluorescent protein (GFP) expression was driven by this regulatory region, the resulting pattern was distinct from that of ace-1. This latter gene is expressed in all body-wall and vulval muscle cells (Culetto et al., 1999), whereas ace-2 is expressed almost exclusively in neurons. ace-3 and ace-4 genes are located in close proximity on chromosome II (Combes et al., 2000). These two genes were first transcribed in vivo as a bicistronic messenger and thus constitute an ace-3;ace-4 operon. However, there was a very low level of monocistronic mRNA of ace-4 (the upstream gene) in vivo, and no ACE-4 enzymatic activity was ever detected. GFP expression driven by a 5' upstream region of the ace-3;ace-4 operon was detected in several muscle cells of the pharynx (pm3, pm4, pm5 and pm7) and in the two canal associated neurons (CAN cells). A dorsal row of body-wall muscle cells was intensively labelled in larval stages but no longer detected in adults. The distinct tissue-specific expression of ace-1, ace-2 and ace-3 (coexpressed only in pm5 cells) indicates that ace genes are not redundant.

  2. Inhibition of acetylcholinesterase and cytochrome oxidase activity in Fasciola gigantica cercaria by phytoconstituents.

    Science.gov (United States)

    Sunita, Kumari; Habib, Maria; Kumar, P; Singh, Vinay Kumar; Husain, Syed Akhtar; Singh, D K

    2016-02-01

    Fasciolosis is an important cattle and human disease caused by Fasciola hepatica and Fasciola gigantica. One of the possible methods to control this problem is to interrupt the life cycle of Fasciola by killing its larva (redia and cercaria) in host snail. Molecular identification of cercaria larva of F. gigantica was done by comparing the nucleotide sequencing with adult F. gigantica. It was noted that nucleotide sequencing of cercaria larva and adult F. gigantica were 99% same. Every month during the year 2011-2012, in vivo treatment with 60% of 4 h LC50 of phyto cercaricides citral, ferulic acid, umbelliferone, azadirachtin and allicin caused significant inhibition of acetylcholinesterase (AChE) and cytochrome oxidase activity in the treated cercaria larva of F. gigantica. Whereas, activity of both enzymes were not significantly altered in the nervous tissues of vector snail Lymnaea acuminata exposed to same treatments. Maximum reduction in AChE (1.35% of control in month of June) and cytochrome oxidase (3.71% of control in the month of July) activity were noted in the cercaria exposed to 60% of 4 h LC50 of azadirachtin and allicin, respectively. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Amperometric Acetylcholinesterase Biosensor Based on Multilayer Multiwall Carbon Nanotubes-chitosan Composite

    Directory of Open Access Journals (Sweden)

    Xia SUN

    2011-11-01

    Full Text Available A simple method for immobilization of acetylcholinesterase (AChE onto the glassy carbon electrode (GCE modified with five layers of multiwall carbon nanotubes (MWNTs-chitosan (CHIT composite was proposed, and thus a fast, sensitive and stable amperometric sensor for quantitative determination of pesticides was developed. Five layers of MWNTs-CHIT promoted electron transfer reactions at a lower potential and catalyzed the electro-oxidation of thiocholine, thus, it improved the detection sensitivity of biosensor. Based on the inhibition of pesticides to the enzymatic activity of AChE, using carbofuran as a model compound, under optimal conditions, the inhibition of carbofuran was proportional to its concentration in two ranges, from 5×10-4 to 7.5 μg/mL and 7.5 to 20 μg/mL with a detection limit of 1×10-4 μg/mL. The constructed biosensor showed prominent characteristics and performances such as good precision, acceptable stability, fast response and low detection limit, which provided a new promising tool for pesticide analysis.

  4. Nanoparticle-based immunosensor with apoferritin templated metallic phosphate label for quantification of phosphorylated acetylcholinesterase

    Energy Technology Data Exchange (ETDEWEB)

    Du, Dan; Chen, Aiqiong; Xie, Yunying; Zhang, Aidong; Lin, Yuehe

    2011-05-15

    A new sandwich-like electrochemical immunosensor has been developed for quantification of organophosphorylated acetylcholinesterase (OP-AChE), an exposure biomarker of organophosphate pesticides and nerve agents. Zirconia nanoparticles (ZrO2 NPs) were anchored on a screen printed electrode (SPE) to preferably capture OP-AChE adducts by metal chelation with phospho-moieties, which was selectively recognized by lead phosphate-apoferritin labeled anti-AChE antibody (LPA-anti-AChE). The sandwich-like immunoreactions were performed among ZrO2 NPs, OP-AChE and LPA-anti-AChE to form ZrO2/OP-AChE/LPA-anti-AChE complex and the released lead ions were detected on a disposable SPE. The binding affinity was investigated by both square wave voltammetry (SWV) and quartz crystal microbalance (QCM) measurements. The proposed immunosensor yielded a linear response current over a broad OP-AChE concentrations range from 0.05 nM to 10 nM, with detection limit of 0.02 nM, which has enough sensitivity for monitoring of low-dose exposure to OPs. This method avoids the drawback of unavailability of commercial OP-specific antibody as well as amplifies detection signal by using apoferritin encoded metallic phosphate nanoparticle tags. This nanoparticle-based immunosensor offers a new method for rapid, sensitive, selective and inexpensive quantification of phosphorylated adducts for monitoring of OP pesticides and nerve agents exposures.

  5. Simplified methods for in vivo measurement of acetylcholinesterase activity in rodent brain

    International Nuclear Information System (INIS)

    Kilbourn, Michael R.; Sherman, Phillip S.; Snyder, Scott E.

    1999-01-01

    Simplified methods for in vivo studies of acetylcholinesterase (AChE) activity in rodent brain were evaluated using N-[ 11 C]methylpiperidinyl propionate ([ 11 C]PMP) as an enzyme substrate. Regional mouse brain distributions were determined at 1 min (representing initial brain uptake) and 30 min (representing trapped product) after intravenous [ 11 C]PMP administration. Single time point tissue concentrations (percent injected dose/gram at 30 min), tissue concentration ratios (striatum/cerebellum and striatum/cortex ratios at 30 min), and regional tissue retention fractions (defined as percent injected dose 30 min/percent injected dose 1 min) were evaluated as measures of AChE enzymatic activity in mouse brain. Studies were carried out in control animals and after dosing with phenserine, a selective centrally active AChE inhibitor; neostigmine, a peripheral cholinesterase inhibitor; and a combination of the two drugs. In control and phenserine-treated animals, absolute tissue concentrations and regional retention fractions provide good measures of dose-dependent inhibition of brain AChE; tissue concentration ratios, however, provide erroneous conclusions. Peripheral inhibition of cholinesterases, which changes the blood pharmacokinetics of the radiotracer, diminishes the sensitivity of all measures to detect changes in central inhibition of the enzyme. We conclude that certain simple measures of AChE hydrolysis rates for [ 11 C]PMP are suitable for studies where alterations of the peripheral blood metabolism of the tracer are kept to a minimum

  6. Simplified methods for in vivo measurement of acetylcholinesterase activity in rodent brain

    Energy Technology Data Exchange (ETDEWEB)

    Kilbourn, Michael R. E-mail: mkilbour@umich.edu; Sherman, Phillip S.; Snyder, Scott E

    1999-07-01

    Simplified methods for in vivo studies of acetylcholinesterase (AChE) activity in rodent brain were evaluated using N-[{sup 11}C]methylpiperidinyl propionate ([{sup 11}C]PMP) as an enzyme substrate. Regional mouse brain distributions were determined at 1 min (representing initial brain uptake) and 30 min (representing trapped product) after intravenous [{sup 11}C]PMP administration. Single time point tissue concentrations (percent injected dose/gram at 30 min), tissue concentration ratios (striatum/cerebellum and striatum/cortex ratios at 30 min), and regional tissue retention fractions (defined as percent injected dose 30 min/percent injected dose 1 min) were evaluated as measures of AChE enzymatic activity in mouse brain. Studies were carried out in control animals and after dosing with phenserine, a selective centrally active AChE inhibitor; neostigmine, a peripheral cholinesterase inhibitor; and a combination of the two drugs. In control and phenserine-treated animals, absolute tissue concentrations and regional retention fractions provide good measures of dose-dependent inhibition of brain AChE; tissue concentration ratios, however, provide erroneous conclusions. Peripheral inhibition of cholinesterases, which changes the blood pharmacokinetics of the radiotracer, diminishes the sensitivity of all measures to detect changes in central inhibition of the enzyme. We conclude that certain simple measures of AChE hydrolysis rates for [{sup 11}C]PMP are suitable for studies where alterations of the peripheral blood metabolism of the tracer are kept to a minimum.

  7. Endosulfan induces changes in spontaneous swimming activity and acetylcholinesterase activity of Jenynsia multidentata (Anablepidae, Cyprinodontiformes)

    Energy Technology Data Exchange (ETDEWEB)

    Ballesteros, M.L. [Facultad de Ciencias Exactas, Fisicas y Naturales, Catedra Diversidad Animal II, Universidad Nacional de Cordoba, Av. Velez Sarsfield 299, 5000 Cordoba (Argentina); Durando, P.E. [Facultad de Ciencias Exactas, Fisicas y Naturales, Departamento de Biologia, Catedra de Fisiologia Animal, Universidad Nacional de San Juan, Complejo ' Islas Malvinas' , Av. Jose I. de la Roza y Meglioli, Rivadavia, San Juan (Argentina); Nores, M.L. [Facultad de Ciencias Medicas, Universidad Nacional de Cordoba-CONICET, Ciudad Universitaria, Cordoba (Argentina); Diaz, M.P. [Facultad de Ciencias Medicas, Catedra de Estadistica y Bioestadistica, Escuela de Nutricion, Universidad Nacional de Cordoba, Pabellon Chile, Ciudad Universitaria, 5000 Cordoba (Argentina); Bistoni, M.A., E-mail: mbistoni@com.uncor.ed [Facultad de Ciencias Exactas, Fisicas y Naturales, Catedra Diversidad Animal II, Universidad Nacional de Cordoba, Av. Velez Sarsfield 299, 5000 Cordoba (Argentina); Wunderlin, D.A. [Facultad de Ciencias Quimicas, Dto. Bioquimica Clinica-CIBICI, Universidad Nacional de Cordoba-CONICET, Haya de la Torre esq. Medina Allende, Ciudad Universitaria, 5000 Cordoba (Argentina)

    2009-05-15

    We assessed changes in spontaneous swimming activity and acetylcholinesterase (AchE) activity of Jenynsia multidentata exposed to Endosulfan (EDS). Females of J. multidentata were exposed to 0.072 and 1.4 mug L{sup -1} EDS. Average speed and movement percentage were recorded during 48 h. We also exposed females to EDS at five concentrations between 0.072 and 1.4 mug L{sup -1} during 24 h, and measured the AchE activity in brain and muscle. At 0.072 mug L{sup -1} EDS swimming motility decreased relative to the control group after 45 h, while at 1.4 mug L{sup -1} EDS swimming motility decreased after 24 h. AchE activity significantly decreased in muscle when J. multidentata were exposed to EDS above 0.072 mug L{sup -1}, while no significant changes were observed in brain. Thus, changes in swimming activity and AchE activity in muscle are good biomarkers of exposure to EDS in J. multidentata. - This work reports changes observed in spontaneous swimming activity and AchE activity of Jenynsia multidentata exposed to sublethal concentrations of Endosulfan.

  8. Diallyl tetrasulfide improves cadmium induced alterations of acetylcholinesterase, ATPases and oxidative stress in brain of rats

    International Nuclear Information System (INIS)

    Pari, Leelavinothan; Murugavel, Ponnusamy

    2007-01-01

    Cadmium (Cd) is a neurotoxic metal, which induces oxidative stress and membrane disturbances in nerve system. The garlic compound diallyl tetrasulfide (DTS) has the cytoprotective and antioxidant activity against Cd induced toxicity. The present study was carried out to investigate the efficacy of DTS in protecting the Cd induced changes in the activity of acetylcholinesterase (AChE), membrane bound enzymes, lipid peroxidation (LPO) and antioxidant status in the brain of rats. In rats exposed to Cd (3 mg/kg/day subcutaneously) for 3 weeks, a significant (P + K + -ATPase, Mg 2+ -ATPase and Ca 2+ -ATPase) were observed in brain tissue. Oral administration of DTS (40 mg/kg/day) with Cd significantly (P < 0.05) diminished the levels of LPO and protein carbonyls and significantly (P < 0.05) increased the activities of ATPases, antioxidant enzymes, GSH and TSH in brain. These results indicate that DTS attenuate the LPO and alteration of antioxidant and membrane bound enzymes in Cd exposed rats, which suggest that DTS protects the brain function from toxic effects of Cd

  9. Characterization of Lignanamides from Hemp (Cannabis sativa L.) Seed and Their Antioxidant and Acetylcholinesterase Inhibitory Activities.

    Science.gov (United States)

    Yan, Xiaoli; Tang, Jiajing; dos Santos Passos, Carolina; Nurisso, Alessandra; Simões-Pires, Claudia Avello; Ji, Mei; Lou, Hongxiang; Fan, Peihong

    2015-12-16

    Hemp seed is known for its content of fatty acids, proteins, and fiber, which contribute to its nutritional value. Here we studied the secondary metabolites of hemp seed aiming at identifying bioactive compounds that could contribute to its health benefits. This investigation led to the isolation of 4 new lignanamides, cannabisin M (2), cannabisin N (5), cannabisin O (8), and 3,3'-demethyl-heliotropamide (10), together with 10 known lignanamides, among which 4 was identified for the first time from hemp seed. Structures were established on the basis of NMR, HR-MS, UV, and IR as well as by comparison with the literature data. Lignanamides 2, 7, and 9-14 showed good antioxidant activity, among which 7, 10, and 13 also inhibited acetylcholinesterase in vitro. The newly identified compounds in this study add to the diversity of hemp seed composition, and the bioassays implied that hemp seed, with lignanamides as nutrients, may be a good source of bioactive and protective compounds.

  10. Synthesis and biological evaluation of lycorine derivatives as dual inhibitors of human acetylcholinesterase and butyrylcholinesterase

    Directory of Open Access Journals (Sweden)

    Wang Yue-Hu

    2012-09-01

    Full Text Available Abstract Background Alzheimer’s disease (AD is a neurologically degenerative disorder that affects more than 20 million people worldwide. The selective butyrylcholinesterase (BChE inhibitors and bivalent cholinesterase (ChE inhibitors represent new treatments for AD. Findings A series of lycorine derivatives (1–10 were synthesized and evaluated for anti-cholinesterase activity. Result showed that the novel compound 2-O-tert-butyldimethylsilyl-1-O-(methylthiomethyllycorine (7 was a dual inhibitor of human acetylcholinesterase (hAChE and butyrylcholinesterase (hBChE with IC50 values of 11.40 ± 0.66 μM and 4.17 ± 0.29 μM, respectively. The structure-activity relationships indicated that (i the 1-O-(methylthiomethyl substituent in lycorine was better than the 1-O-acetyl group for the inhibition of cholinesterase; (ii the acylated or etherified derivatives of lycorine and lycorin-2-one were more potent against hBChE than hAChE; and (iii the oxidation of lycorine at C-2 decreases the activity. Conclusion Acylated or etherified derivatives of lycorine are potential dual inhibitors of hBChE and hAChE. Hence, further study on the modification of lycorine for ChE inhibition is necessary.

  11. Carbon-11 labelling of an inhibitor of acetylcholinesterase: [11C]physostigmine

    International Nuclear Information System (INIS)

    Bonnot-Lours, S.; Crouzel, C.; Prenant, C.; Hinnen, F.

    1993-01-01

    Physostigmine, an alkaloid from calabar bean is a strong inhibitor of acetylcholinesterase and has been used clinically in the treatment of glaucoma, atropine intoxication, myasthenia gravis and more recently, in experimental trials in Alzheimer's disease. In order to study the AChE activity in the brain by positron emission tomography, we have undertaken the labelling of physostigmine with carbon-11. The synthesis involves the reaction of [ 11 C]methylisocyanate with eseroline. [ 11 C]Methylisocyanate was obtained by heating [ 11 C]acetylchloride with tetrabutylammonium azide in toluene. The synthesis of [ 11 C]CH 3 COC1 involves the carbonation of methylmagnesium bromide in THF with cyclotron produced [ 11 C]carbon dioxide and the addition of phthaloyl dichloride. The [ 11 C]methylisocyanate is distilled into a solution of eseroline in ether with a small piece of sodium. After 10 minutes at 25 o C, the solution is purified by HPLC and the appropriate fraction collected. Starting with 55.5 GBq (1.5 Ci) of [ 11 C]carbon dioxide, 0.92-1.48 GBq (25-40 mCi) of [ 11 C]Physostigmine are obtained 57 minutes after EOB. (author)

  12. Acetylcholinesterase activity in seabirds affected by the Prestige oil spill on the Galician coast (Spain)

    Energy Technology Data Exchange (ETDEWEB)

    Oropesa, Ana-Lourdes; Perez-Lopez, Marcos; Hernandez, David; Soler, Francisco [Toxicology Area, Faculty of Veterinary Science (UEX), Avda. de la Universidad s/n. 10071 Caceres (Spain); Garcia, Jesus-Pablo [Toxicology Area, National Centre of Environmental Health, Instituto de Salud Carlos III, Majadahonda, Madrid (Spain); Fidalgo, Luis-Eusebio; Lopez-Beceiro, Ana [Rof Codina Clinical Hospital, Faculty of Veterinary Science (USC), Estrada de Granxa s/n. 27003 Lugo (Spain)

    2007-01-01

    In November 2002, the tanker Prestige broke in two and sank at the bottom of the ocean spilling about 70,000 t of fuel oil, which reached the coast of Galicia. It was considered the largest spill in maritime history, greatly affecting marine and related avian species. The spilled fuel oil contained high concentrations of polycyclic aromatic hydrocarbons (PAHs). Many species were affected and were found dead, although ongoing research is still being carried out on the sublethal effects. In this sense, little is known about the action of PAHs on Cholinesterase activity in seabirds. Consequently, the purpose of this study was to provide more information on the neurotoxicity of fuel oil on the seabirds most affected by the Prestige accident: common guillemot, Atlantic puffin and razorbill. On the other hand, data on normal values of acetylcholinesterase (AChE) activity were obtained to supply non-exposed values in seabirds. The oil spill produced a clear inhibitory effect on brain AChE activity in common guillemot (16%, p {<=} 0.01) and razorbill (22%, p {<=} 0.01), but not in Atlantic puffin (4%). Physiological levels of brain AChE, expressed in nmol acetylcholine hydrolysed min{sup -} {sup 1} mg{sup -} {sup 1} protein were similar in non-exposed common guillemot (388.6 {+-} 95.0) and Atlantic puffin (474.0 {+-} 60.7), however, razorbill values were higher (644.6 {+-} 66.9). (author)

  13. Blocked Enzymatic Etching of Gold Nanorods: Application to Colorimetric Detection of Acetylcholinesterase Activity and Its Inhibitors.

    Science.gov (United States)

    Saa, Laura; Grinyte, Ruta; Sánchez-Iglesias, Ana; Liz-Marzán, Luis M; Pavlov, Valeri

    2016-05-04

    The anisotropic morphology of gold nanorods (AuNRs) has been shown to lead to nonuniform ligand distribution and preferential etching through their tips. We have recently demonstrated that this effect can be achieved by biocatalytic oxidation with hydrogen peroxide, catalyzed by the enzyme horseradish peroxidase (HRP). We report here that modification of AuNRs with thiol-containing organic molecules such as glutathione and thiocholine hinders enzymatic AuNR etching. Higher concentrations of thiol-containing molecules in the reaction mixture gradually decrease the rate of enzymatic etching, which can be monitored by UV-vis spectroscopy through changes in the AuNR longitudinal plasmon band. This effect can be applied to develop novel optical assays for acetylcholinesterase (AChE) activity. The biocatalytic hydrolysis of acetylthiocholine by AChE yields thiocholine, which prevents enzymatic AuNR etching in the presence of HRP. Additionally, the same bioassay can be used for the detection of nanomolar concentrations of AChE inhibitors such as paraoxon and galanthamine.

  14. Acetylcholinesterase in Biofouling Species: Characterization and Mode of Action of Cyanobacteria-Derived Antifouling Agents.

    Science.gov (United States)

    Almeida, Joana R; Freitas, Micaela; Cruz, Susana; Leão, Pedro N; Vasconcelos, Vitor; Cunha, Isabel

    2015-07-24

    Effective and ecofriendly antifouling (AF) compounds have been arising from naturally produced chemicals. The objective of this study is to use cyanobacteria-derived agents to investigate the role of acetylcholinesterase (AChE) activity as an effect and/or mode of action of promising AF compounds, since AChE inhibitors were found to inhibit invertebrate larval settlement. To pursue this objective, in vitro quantification of AChE activity under the effect of several cyanobacterial strain extracts as potential AF agents was performed along with in vivo AF (anti-settlement) screening tests. Pre-characterization of different cholinesterases (ChEs) forms present in selected tissues of important biofouling species was performed to confirm the predominance of AChE, and an in vitro AF test using pure AChE activity was developed. Eighteen cyanobacteria strains were tested as source of potential AF and AChE inhibitor agents. Results showed effectiveness in selecting promising eco-friendly AF agents, allowing the understanding of the AF biochemical mode of action induced by different compounds. This study also highlights the potential of cyanobacteria as source of AF agents towards invertebrate macrofouling species.

  15. Acetylcholinesterase in Biofouling Species: Characterization and Mode of Action of Cyanobacteria-Derived Antifouling Agents

    Directory of Open Access Journals (Sweden)

    Joana R. Almeida

    2015-07-01

    Full Text Available Effective and ecofriendly antifouling (AF compounds have been arising from naturally produced chemicals. The objective of this study is to use cyanobacteria-derived agents to investigate the role of acetylcholinesterase (AChE activity as an effect and/or mode of action of promising AF compounds, since AChE inhibitors were found to inhibit invertebrate larval settlement. To pursue this objective, in vitro quantification of AChE activity under the effect of several cyanobacterial strain extracts as potential AF agents was performed along with in vivo AF (anti-settlement screening tests. Pre-characterization of different cholinesterases (ChEs forms present in selected tissues of important biofouling species was performed to confirm the predominance of AChE, and an in vitro AF test using pure AChE activity was developed. Eighteen cyanobacteria strains were tested as source of potential AF and AChE inhibitor agents. Results showed effectiveness in selecting promising eco-friendly AF agents, allowing the understanding of the AF biochemical mode of action induced by different compounds. This study also highlights the potential of cyanobacteria as source of AF agents towards invertebrate macrofouling species.

  16. An Unusual Dimeric Inhibitor of Acetylcholinesterase: Cooperative Binding of Crystal Violet

    Directory of Open Access Journals (Sweden)

    Anders Allgardsson

    2017-08-01

    Full Text Available Acetylcholinesterase (AChE is an essential enzyme that terminates cholinergic transmission by a rapid hydrolysis of the neurotransmitter acetylcholine. AChE is an important target for treatment of various cholinergic deficiencies, including Alzheimer’s disease and myasthenia gravis. In a previous high throughput screening campaign, we identified the dye crystal violet (CV as an inhibitor of AChE. Herein, we show that CV displays a significant cooperativity for binding to AChE, and the molecular basis for this observation has been investigated by X-ray crystallography. Two monomers of CV bind to residues at the entrance of the active site gorge of the enzyme. Notably, the two CV molecules have extensive intermolecular contacts with each other and with AChE. Computational analyses show that the observed CV dimer is not stable in solution, suggesting the sequential binding of two monomers. Guided by the structural analysis, we designed a set of single site substitutions, and investigated their effect on the binding of CV. Only moderate effects on the binding and the cooperativity were observed, suggesting a robustness in the interaction between CV and AChE. Taken together, we propose that the dimeric cooperative binding is due to a rare combination of chemical and structural properties of both CV and the AChE molecule itself.

  17. Endosulfan induces changes in spontaneous swimming activity and acetylcholinesterase activity of Jenynsia multidentata (Anablepidae, Cyprinodontiformes)

    International Nuclear Information System (INIS)

    Ballesteros, M.L.; Durando, P.E.; Nores, M.L.; Diaz, M.P.; Bistoni, M.A.; Wunderlin, D.A.

    2009-01-01

    We assessed changes in spontaneous swimming activity and acetylcholinesterase (AchE) activity of Jenynsia multidentata exposed to Endosulfan (EDS). Females of J. multidentata were exposed to 0.072 and 1.4 μg L -1 EDS. Average speed and movement percentage were recorded during 48 h. We also exposed females to EDS at five concentrations between 0.072 and 1.4 μg L -1 during 24 h, and measured the AchE activity in brain and muscle. At 0.072 μg L -1 EDS swimming motility decreased relative to the control group after 45 h, while at 1.4 μg L -1 EDS swimming motility decreased after 24 h. AchE activity significantly decreased in muscle when J. multidentata were exposed to EDS above 0.072 μg L -1 , while no significant changes were observed in brain. Thus, changes in swimming activity and AchE activity in muscle are good biomarkers of exposure to EDS in J. multidentata. - This work reports changes observed in spontaneous swimming activity and AchE activity of Jenynsia multidentata exposed to sublethal concentrations of Endosulfan.

  18. Carbon-11 labelling of an inhibitor of acetylcholinesterase: [[sup 11]C]physostigmine

    Energy Technology Data Exchange (ETDEWEB)

    Bonnot-Lours, S.; Crouzel, C.; Prenant, C.; Hinnen, F. (CEA, 91 - Orsay (France). Service Hospitalier Frederic Joliot)

    1993-01-01

    Physostigmine, an alkaloid from calabar bean is a strong inhibitor of acetylcholinesterase and has been used clinically in the treatment of glaucoma, atropine intoxication, myasthenia gravis and more recently, in experimental trials in Alzheimer's disease. In order to study the AChE activity in the brain by positron emission tomography, we have undertaken the labelling of physostigmine with carbon-11. The synthesis involves the reaction of [[sup 11]C]methylisocyanate with eseroline. [[sup 11]C]Methylisocyanate was obtained by heating [[sup 11]C]acetylchloride with tetrabutylammonium azide in toluene. The synthesis of [[sup 11]C]CH[sub 3]COC1 involves the carbonation of methylmagnesium bromide in THF with cyclotron produced [[sup 11]C]carbon dioxide and the addition of phthaloyl dichloride. The [[sup 11]C]methylisocyanate is distilled into a solution of eseroline in ether with a small piece of sodium. After 10 minutes at 25[sup o]C, the solution is purified by HPLC and the appropriate fraction collected. Starting with 55.5 GBq (1.5 Ci) of [[sup 11]C]carbon dioxide, 0.92-1.48 GBq (25-40 mCi) of [[sup 11]C]Physostigmine are obtained 57 minutes after EOB. (author).

  19. A method for acetylcholinesterase staining of brain sections previously processed for receptor autoradiography.

    Science.gov (United States)

    Lim, M M; Hammock, E A D; Young, L J

    2004-02-01

    Receptor autoradiography using selective radiolabeled ligands allows visualization of brain receptor distribution and density on film. The resolution of specific brain regions on the film often can be difficult to discern owing to the general spread of the radioactive label and the lack of neuroanatomical landmarks on film. Receptor binding is a chemically harsh protocol that can render the tissue virtually unstainable by Nissl and other conventional stains used to delineate neuroanatomical boundaries of brain regions. We describe a method for acetylcholinesterase (AChE) staining of slides previously processed for receptor binding. AChE staining is a useful tool for delineating major brain nuclei and tracts. AChE staining on sections that have been processed for receptor autoradiography provides a direct comparison of brain regions for more precise neuroanatomical description. We report a detailed thiocholine protocol that is a modification of the Koelle-Friedenwald method to amplify the AChE signal in brain sections previously processed for autoradiography. We also describe several temporal and experimental factors that can affect the density and clarity of the AChE signal when using this protocol.

  20. Formulation and characterization of novel functional beverages with antioxidant and anti-acetylcholinesterase activities

    Directory of Open Access Journals (Sweden)

    Suree Nanasombat

    2015-01-01

    beverages B1, B2, B3, B4 and B5 in the ratio of 60:40 to prepare alcoholic beverages W1, W2, W3, W4 and W5, respectively. Two different fermentation conditions (fermentation with or without pieces of sliced medicinal plant residue, PMPR were compared. After fermenting, racking and aging, all alcoholic beverages, as well as all non-alcoholic beverages,were analyzed for some phytochemical properties. Results: Grape fermented with PMPR had higher anti-acetylcholinesterase and antioxidant activities, and total phenolics, flavonoids and tannins, compared to the others. Among all nonalcoholic beverages, the beverage B3 contained the highest anti-acetylcholinesterase (22.78% inhibition at 1:10,000 dilution and antioxidant activities (reducing capacity, 4.22 mmol Fe(II/100 mL, total phenolics, flavonoids, and tannins (494.44 mg gallic acid equivalents (GAE, 383.22 mg catechin equivalents (CE and 338.29 mg tannic acid equivalents ((TAE/100 mL, respectively. Among all alcoholic beverages, the beverage W3 (fermented with PMPR exhibited the highest antioxidant activity (DPPH radical inhibition, 95.99 mg trolox equivalents and reducing capacity, 3.57 mmol Fe(II /100 mL, total phenolics, flavonoids and tannins (239.71 mg GAE, 372.67 mg CE and 157.67 mg TAE/100 mL, respectively. The beverage W2 (fermented with PMPR had the highest anti-acetylcholinesterase activity (21.35% inhibition at 1:10,000 dilution. Conclusion: The beverages B3, W2 and W3 contained valuable sources of natural antioxidants and acetylcholinesterase inhibitors, and may provide health benefits when consumed.

  1. Surface display of recombinant Drosophila melanogaster acetylcholinesterase for detection of organic phosphorus and carbamate pesticides.

    Directory of Open Access Journals (Sweden)

    Jingquan Li

    Full Text Available Acetylcholinesterase (AChE is commonly used for the detection of organophosphate (OP and carbamate (CB insecticides. However, the cost of this commercially available enzyme is high, making high-throughput insecticide detection improbable. In this study we constructed a new AChE yeast expression system in Saccharomyces cerevisiae for the expression of a highly reactive recombinant AChE originating from Drosophila melanogaster (DmAChE. Specifically, the coding sequence of DmAChE was fused with the 3'-terminal half of an α-agglutinin anchor region, along with an antigen tag for the detection of the recombinant protein. The target sequence was cloned into the yeast expression vector pYes-DEST52, and the signal peptide sequence was replaced with a glucoamylase secretion region for induced expression. The resultant engineered vector was transformed into S. cerevisiae. DmAChE was expressed and displayed on the cell surface after galactose induction. Our results showed that the recombinant protein displayed activity comparable to the commercial enzyme. We also detected different types of OP and CB insecticides through enzyme inhibition assays, with the expressed DmAChE showing high sensitivity. These results show the construction of a new yeast expression system for DmAChE, which can subsequently be used for detecting OP and CB insecticides with reduced economic costs.

  2. Molecular evaluation of herbal compounds as potent inhibitors of acetylcholinesterase for the treatment of Alzheimer's disease.

    Science.gov (United States)

    Chen, Yan-Xiu; Li, Guan-Zeng; Zhang, Bin; Xia, Zhang-Yong; Zhang, Mei

    2016-07-01

    Alzheimer's disease (AD) is a progressive disease and the predominant cause of dementia. Common symptoms include short-term memory loss, and confusion with time and place. Individuals with AD depend on their caregivers for assistance, and may pose a burden to them. The acetylcholinesterase (AChE) enzyme is a key target in AD and inhibition of this enzyme may be a promising strategy in the drug discovery process. In the present study, an inhibitory assay was carried out against AChE using total alkaloidal plants and herbal extracts commonly available in vegetable markets. Subsequently, molecular docking simulation analyses of the bioactive compounds present in the plants were conducted, as well as a protein‑ligand interaction analysis. The stability of the docked protein‑ligand complex was assessed by 20 ns molecular dynamics simulation. The inhibitory assay demonstrated that Uncaria rhynchophylla and Portulaca oleracea were able to inhibit AChE. In addition, molecular docking simulation analyses indicated that catechin present in Uncaria rhynchophylla, and dopamine and norepinephrine present in Portulaca oleracea, had the best docking scores and interaction energy. In conclusion, catechin in Uncaria rhynchophylla, and dopamine and norepinephrine in Portulaca oleracea may be used to treat AD.

  3. Plant-derived acetylcholinesterase inhibitory alkaloids for the treatment of Alzheimer's disease

    Directory of Open Access Journals (Sweden)

    Dall'Acqua S

    2013-01-01

    Full Text Available Stefano Dall'AcquaDepartment of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, ItalyAbstract: The inhibition of acetylcholinesterase (AChE has been one of the most used strategies for the treatment of Alzheimer's disease (AD. The AChE inhibitors (AChE-I produce not only short-term symptomatic effects, but can also play a role in other pathological mechanisms of the disease (eg, formation of amyloid-β plaques, which has renewed interest in the discovery of such inhibitors. Four of the five currently prescribed treatments for AD are AChE-I. Natural alkaloids such as galantamine or alkaloid-related synthetic compounds (such as rivastigmine are considered beneficial for patients with mild-to-moderate AD. However, there is a need for the discovery of more effective compounds and for this reason, plants can still be a potential source of new AChE-I. Findings and advances in knowledge about natural alkaloids as potential new drugs acting as AChE-I will be summarized in this paper.Keywords: quinolizidine, steroidal, indole, isoquinoline

  4. A sensitive acetylcholinesterase biosensor based on gold nanorods modified electrode for detection of organophosphate pesticide.

    Science.gov (United States)

    Lang, Qiaolin; Han, Lei; Hou, Chuantao; Wang, Fei; Liu, Aihua

    2016-08-15

    A sensitive amperometric acetylcholinesterase (AChE) biosensor, based on gold nanorods (AuNRs), was developed for the detection of organophosphate pesticide. Compared with Au@Ag heterogeneous NRs, AuNRs exhibited excellent electrocatalytic properties, which can electrocatalytically oxidize thiocholine, the hydrolysate of acetylthiocholine chloride (ATCl) by AChE at +0.55V (vs. SCE). The AChE/AuNRs/GCE biosensor was fabricated on basis of the inhibition of AChE activity by organophosphate pesticide. The biosensor could detect paraoxon in the linear range from 1nM to 5μM and dimethoate in the linear range from 5nM to 1μM, respectively. The detection limits of paraoxon and dimethoate were 0.7nM and 3.9nM, which were lower than the reported AChE biosensor. The proposed biosensor could restore to over 95% of its original current, which demonstrated the good reactivation. Moreover, the biosensor can be applicable to real water sample measurement. Thus, the biosensor exhibited low applied potential, high sensitivity and good stability, providing a promising tool for analysis of pesticides. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Acetylcholinesterase triggers the aggregation of PrP 106-126

    International Nuclear Information System (INIS)

    Pera, M.; Roman, S.; Ratia, M.; Camps, P.; Munoz-Torrero, D.; Colombo, L.; Manzoni, C.; Salmona, M.; Badia, A.; Clos, M.V.

    2006-01-01

    Acetylcholinesterase (AChE), a senile plaque component, promotes amyloid-β-protein (Aβ) fibril formation in vitro. The presence of prion protein (PrP) in Alzheimer's disease (AD) senile plaques prompted us to assess if AChE could trigger the PrP peptides aggregation as well. Consequently, the efficacy of AChE on the PrP peptide spanning-residues 106-126 aggregation containing a coumarin fluorescence probe (coumarin-PrP 106-126) was studied. Kinetics of coumarin-PrP 106-126 aggregation showed a significant increase of maximum size of aggregates (MSA), which was dependent on AChE concentration. AChE-PrP 106-126 aggregates showed the tinctorial and optical amyloid properties as determined by polarized light and electronic microscopy analysis. A remarkable inhibition of MSA was obtained with propidium iodide, suggesting that AChE triggers PrP 106-126 and Aβ aggregation through a similar mechanism. Huprines (AChE inhibitors) also significantly decreased MSA induced by AChE as well, unveiling the potential interest for some AChE inhibitors as a novel class of potential anti-prion drugs

  6. An Acetylcholinesterase-Based Chronoamperometric Biosensor for Fast and Reliable Assay of Nerve Agents

    Directory of Open Access Journals (Sweden)

    Rene Kizek

    2013-08-01

    Full Text Available The enzyme acetylcholinesterase (AChE is an important part of cholinergic nervous system, where it stops neurotransmission by hydrolysis of the neurotransmitter acetylcholine. It is sensitive to inhibition by organophosphate and carbamate insecticides, some Alzheimer disease drugs, secondary metabolites such as aflatoxins and nerve agents used in chemical warfare. When immobilized on a sensor (physico-chemical transducer, it can be used for assay of these inhibitors. In the experiments described herein, an AChE- based electrochemical biosensor using screen printed electrode systems was prepared. The biosensor was used for assay of nerve agents such as sarin, soman, tabun and VX. The limits of detection achieved in a measuring protocol lasting ten minutes were 7.41 × 10−12 mol/L for sarin, 6.31 × 10−12 mol /L for soman, 6.17 × 10−11 mol/L for tabun, and 2.19 × 10−11 mol/L for VX, respectively. The assay was reliable, with minor interferences caused by the organic solvents ethanol, methanol, isopropanol and acetonitrile. Isopropanol was chosen as suitable medium for processing lipophilic samples.

  7. Binding free energy calculations to rationalize the interactions of huprines with acetylcholinesterase

    Science.gov (United States)

    Nascimento, Érica C. M.; Oliva, Mónica; Andrés, Juan

    2018-05-01

    In the present study, the binding free energy of a family of huprines with acetylcholinesterase (AChE) is calculated by means of the free energy perturbation method, based on hybrid quantum mechanics and molecular mechanics potentials. Binding free energy calculations and the analysis of the geometrical parameters highlight the importance of the stereochemistry of huprines in AChE inhibition. Binding isotope effects are calculated to unravel the interactions between ligands and the gorge of AChE. New chemical insights are provided to explain and rationalize the experimental results. A good correlation with the experimental data is found for a family of inhibitors with moderate differences in the enzyme affinity. The analysis of the geometrical parameters and interaction energy per residue reveals that Asp72, Glu199, and His440 contribute significantly to the network of interactions between active site residues, which stabilize the inhibitors in the gorge. It seems that a cooperative effect of the residues of the gorge determines the affinity of the enzyme for these inhibitors, where Asp72, Glu199, and His440 make a prominent contribution.

  8. Anti-Acetylcholinesterase and Antioxidant Appraisal of the Bulb Extracts of Five Sternbergia Species

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    Ilkay Erdogan Orhan

    2011-01-01

    Full Text Available In the current study, we examined anti-acetylcholinesterase (AChE and antioxidant activities of the ethyl acetate, methanol, and water extracts from the bulbs of Turkish Sternbergia Waldst. & Kit. (Amaryllidaceae species; S. candida, S. clusiana, S. fisheriana, S. lutea subsp. lutea, and S. lutea subsp. sicula. Anti-AChE activity was tested by spectrophotometric method of Ellman using ELISA microplate reader at 50, 100, and 200 μg mL -1 concentrations. Antioxidant activity of the extracts was evaluated by DPPH radical scavenging activity, ferrous ion-chelating capacity, ferric-reducing antioxidant power, and beta-carotene bleaching assays at 500, 1000, and 2000 μg mL -1. Total phenol and flavonoid contents of the extracts were determined via Folin-Ciocalteau’s and AlCl3 reagents, respectively. The ethyl acetate extract of S. fischeriana was the most active in anti-AChE assay (90.94% and 98.02% of inhibitions at 100 and 200 μg mL -1 concentrations, respectively. Antioxidant activity of the extracts was found to be not significant.

  9. Acetylcholinesterase inhibitory, antioxidant, and antimicrobial activities of Salvia tomentosa Mill. essential oil

    Directory of Open Access Journals (Sweden)

    ANDREY MARCHEV

    2015-08-01

    Full Text Available Chemical composition and bioactivity of essential oil from Salvia tomentosa Mill. natively grown in Bulgaria were investigated. GC-MS analysis identified 60 compounds which represented 98% of the oil constituents. The prevalent constituents were monoterpenes with eight dominant compounds being identified: borneol (10.3%, β-pinene (9%, camphor (7.9%, α-pinene (6%, camphene (4%, 1.8-cineole (3.8%, α-limonene (3.5% and β-caryophyllene (3%. The essential oil showed considerable acetylcholinesterase inhibitory activity (IC50=0.28±0.06 µg/mL, comparable with that of galanthamine. Study of antioxidant activity strongly suggested that the hydrogen atom transfer reaction was preferable over the electron transfer (ORAC=175.0±0.40 µM Trolox equivalents/g oil and FRAP=1.45±0.21 mM Trolox equivalents/g oil. The essential oil showed moderate antifungal and antibacterial activities against Candida albicans and Gram-positive bacteria, whereas it was almost inactive against the investigated Gram-negative strains. The results suggested that the essential oil of Bulgarian S. tomentosa could be considered as a prospective active ingredient for prevention of oxidative stress-related and neurodegenerative disorders in aromatherapy. Because of the high antioxidant capacity, the oil could be considered as natural supplement or antioxidant in cosmetics and food products.

  10. Effect of carbaryl (carbamate insecticide) on acetylcholinesterase activity of two strains of Daphnia magna (Crustacea, Cladocera).

    Science.gov (United States)

    Toumi, Hela; Bejaoui, Mustapha; Touaylia, Samir; Burga Perez, Karen F; Ferard, Jean François

    2016-11-01

    The present study was designed to investigate the effect of carbaryl (carbamate insecticide) on the acetylcholinesterase activity in two strains (same clone A) of the crustacean cladoceran Daphnia magna. Four carbaryl concentrations (0.4, 0.9, 1.8 and 3.7 µg L(-1)) were compared against control AChE activity. Our results showed that after 48 h of carbaryl exposure, all treatments induced a significant decrease of AChE activities whatever the two considered strains. However, different responses were registered in terms of lowest observed effect concentrations (LOEC: 0.4 µg L(-1) for strain 1 and 0.9 µg L(-1) for strains 2) revealing differences in sensitivity among the two tested strains of D. magna. These results suggest that after carbaryl exposure, the AChE activity responses can be also used as a biomarker of susceptibility. Moreover, our results show that strain1 is less sensitive than strain 2 in terms of IC50-48 h of AChE activity. Comparing the EC50-48 h of standard ecotoxicity test and IC50-48 h of AChE inhibition, there is the same order of sensitivity with both strains.

  11. Toxicity of azodrin on the morphology and acetylcholinesterase activity of the earthworm Eisenia foetida

    International Nuclear Information System (INIS)

    Rao, J.V.; Kavitha, P.

    2004-01-01

    The acute toxicity of azodrin (monocrotophos, an organophosphorus insecticide) was determined on a soil organism, Eisenia foetida. The median lethal concentrations (LC 50 ) were derived from a 48-h paper contact test and from artificial soil tests. The LC 50 of azodrin in the paper contact test was 0.46±0.1 μg cm -2 (23±6 mg L -1 ) and those in the 7- and 14-day artificial soil tests were 171±21 and 132±20 mg kg -1 , respectively. The neurotoxic potentiality of azodrin was assessed by using a marker enzyme, acetylcholinesterase (AChE; EC 3.1.1.7) in both in vitro and in vivo experiments. The progressive signs of morphological destruction are correlated with percentage inhibition of AChE in the in vivo experiments. The kinetics of AChE activity in the presence and absence of azodrin indicated that the toxicant is competitive in nature. This study demonstrated that azodrin causes concentration-dependent changes in the morphology and AChE activity of the earthworm E. foetida

  12. Sublethal Effects of Insecticide Exposure on Megacopta cribraria (Fabricius) Nymphs: Key Biological Traits and Acetylcholinesterase Activity.

    Science.gov (United States)

    Miao, Jin; Reisig, Dominic D; Li, Guoping; Wu, Yuqing

    2016-01-01

    Megacopta cribraria F. (Hemiptera: Plataspidae), the kudzu bug, is an invasive insect pest of U.S. soybean. At present, insecticide application is the primary and most effective control option for M. cribraria In this study, the potential effects of sublethal and low-lethal concentrations (LC10 and LC40) of three common insecticides on key biological traits and acetylcholinesterase (AChE) activity of the treated nymphal stage of insect were assessed. The results show that the sublethal concentration of imidacloprid significantly reduced adult emergence rate of M. cribraria A low-lethal concentration of imidacloprid significantly increased nymphal development time, but significantly decreased adult emergence rate and adult longevity. Both sublethal and low-lethal concentrations of acephate caused an increase in nymphal development time and a reduction in adult emergence rate and adult longevity. Fecundity of females was significantly reduced only by exposure to low-lethal concentrations of acephate. Sublethal and low-lethal concentrations of bifenthrin increased nymphal development time, but significantly decreased adult emergence rate. In addition, we found that the AChE activity of M. cribraria was significantly increased only by LC40 imidacloprid, but strongly inhibited by acephate. © The Author 2016. Published by Oxford University Press on behalf of the Entomological Society of America.

  13. Novel acetylcholinesterase inhibitors from Zijuan tea and biosynthetic pathway of caffeoylated catechin in tea plant.

    Science.gov (United States)

    Wang, Wei; Fu, Xi-Wen; Dai, Xin-Long; Hua, Fang; Chu, Gang-Xiu; Chu, Ming-Jie; Hu, Feng-Lin; Ling, Tie-Jun; Gao, Li-Ping; Xie, Zhong-Wen; Wan, Xiao-Chun; Bao, Guan-Hu

    2017-12-15

    Zijuan tea is a special cultivar of Yunnan broad-leaf tea (Camellia sinensis var. assamica) with purple buds, leaves, and stems. Phytochemical study on this tea led to the discovery of three hydroxycinnamoylated catechins (HCCs) (1-3), seven other catechins (4-10), three proanthocyanidins (11-13), five flavones and flavone glycosides (14-18), two alkaloids (19, 20), one steroid (21), and one phenylpropanoid glycoside (22). The isolation and structural elucidation of the caffeoylated catechin (1) by means of spectroscopic techniques were described. We also provide the first evidence that 1 is synthesized via a two-step pathway in tea plant. The three HCCs (1-3) were investigated on their bioactivity through molecular modeling simulation and biochemical experiments. Our results show that they bind acetylcholinesterase (AChE) tightly and have strong AChE inhibitory activity with IC 50 value at 2.49, 11.41, 62.26μM, respectively. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Inhibitory effect of ebselen on cerebral acetylcholinesterase activity in vitro: kinetics and reversibility of inhibition.

    Science.gov (United States)

    Martini, Franciele; Bruning, César Augusto; Soares, Suelen Mendonca; Nogueira, Cristina Wayne; Zeni, Gilson

    2015-01-01

    Ebselen is a synthetic organoselenium compound that has been considered a potential pharmacological agent with low toxicity, showing antioxidant, anti-inflammatory and neuroprotective effects. It is bioavailable, blood-brain barrier permeant and safe based on cellular toxicity and Phase I-III clinical trials. There is evidence that ebselen inhibits acetylcholinesterase (AChE) activity, an enzyme that plays a key role in the cholinergic system by hydrolyzing acetylcholine (ACh), in vitro and ex vivo. This system has a well-known relationship with cognitive process, and AChE inhibitors, such as donepezil and galantamine, have been used to treat cognitive deficits, mainly in the Alzheimer's Disease (AD). However, these drugs have poor bioavailability and a number of side effects, including gastrointestinal upsets and hepatotoxicity. In this way, this study aimed to evaluate the effect of ebselen on cerebral AChE activity in vitro and to determine the kinetic profile and the reversibility of inhibition by dialysis. Ebselen inhibited the cerebral AChE activity with an IC50 of 29 µM, similar to IC50 found with pure AChE from electric eel, demonstrating a mixed and reversible inhibition of AChE, since it increased Km and decreased Vmax. The AChE activity was recovered within 60 min of dialysis. Therefore, the use of ebselen as a therapeutic agent for treatment of AD should be considered, although memory behavior tasks are needed to support such hypothesis.

  15. EFFICIENCY OF REAL-TIME PCR FOR 18S rRNA AMPLIFICATION OF SORBUS DOMESTICA, L.

    Directory of Open Access Journals (Sweden)

    Petronela Poláčeková

    2012-10-01

    Full Text Available Normal 0 21 false false false SK X-NONE X-NONE Nowadays, the awareness is given more and more to underutilized and  unusual fruits. One of them is Sorbus domestica, L. not only as an endangered species, but as well as a promising and economically usable crop. The work was aimed for finding a total genomic DNA isolating methods from fresh plant material and confirmation of the optimized method by the detection of 18S rRNA gene using real-time PCR. Two commercial isolation kits were tested -  Invisorb® Spin Plant Mini Kit and Wizard ® Genomic DNA. Higher purity and yield of DNA isolation kit showed Invisorb kit. The effective and pure PCR amplification was confirmed for Invisorb, too when 20 ng undiluted DNA at annealing temperature of 64.5 °C.doi:10.5219/203

  16. Old Apple (Malus domestica L. Borkh) Varieties with Hypoallergenic Properties: An Integrated Approach for Studying Apple Allergenicity.

    Science.gov (United States)

    Vegro, Mara; Eccher, Giulia; Populin, Francesca; Sorgato, Chiara; Savazzini, Federica; Pagliarani, Giulia; Tartarini, Stefano; Pasini, Gabriella; Curioni, Andrea; Antico, Andrea; Botton, Alessandro

    2016-12-07

    Freshly consumed apples (Malus domestica L. Borkh) can cause allergic reactions because of the presence of four classes of allergens. Knowledge of the genetic factors affecting the allergenic potential of apples would provide important information for the selection of hypoallergenic genotypes, which can be combined with the adoption of new agronomical practices to produce fruits with a reduced amount of allergens. In the present research, a multiple analytical approach was adopted to characterize the allergenic potential of 24 apple varieties released at different ages (pre- and post-green revolution). A specific workflow was set up including protein quantification by means of polyclonal antibodies, immunological analyses with sera of allergic subjects, enzymatic assays, clinical assessments on allergic patients, and gene expression assays on fruit samples. Taken as a whole, the results indicate that most of the less allergenic genotypes were found among those deriving from selection processes carried out prior to the so-called "green revolution".

  17. The effects of gamma irradiation on medicinal plants and spices (3), Curcuma xanthoriza, curcuma aeruginosa, curcuma domestica and kaemferia galanga

    International Nuclear Information System (INIS)

    Chosdu, Rahayu; Hilmy, Nazly; Bagiawati, Sri

    1985-01-01

    This research was conducted to investigate the effects of gamma irradiation on medicinal plants by studying the characteristics of the essential oils of these samples. The samples studied were ''temu lawak'' (Curcuma xanthoriza), ''temu hitam'' (Curcuma aeruginosa), turmeric (Curcuma domestica) and galanga (Kaemferia galanga). The sliced tubers, which contained 8 to 14% of moisture were packed in low density polyethylene bags of 0.13 mm thickness, then irradiated with doses of 0, 5 and 10 kGy. Some samples were directly analyzed and the rest were stored for six months at temperatures of 29±2 0 C and under 70 to 95% humidity. The results showed that the water activity (Aw), the content and characteristics of essential oils, analyzed on a gas liquid chromatography, were not changed by irradiation and storage, but the water content was changed by storage. It was observed also that after irradiation and storage mould and bacteria contents decreased. (author)

  18. Ultraviolet radiation-induced histopathologic changes in the skin of the marsupial Monodelphis domestica. II. Quantitative studies of the photoreactivation of induced hyperplasia and sunburn cell formation

    International Nuclear Information System (INIS)

    Ley, R.D.; Applegate, L.A.

    1985-01-01

    Induction of sunburn cells (SBCs) and hyperplasia of the epidermis of the marsupial Monodelphis domestica by ultraviolet radiation (UVR) has been studied. A dose of 500 J/m2 (approximately 1 minimal erythemal dose) from an FS-40 sunlamp induced measurable numbers of SBCs with a peak number at 32-48 h post-UVR exposure of skin to photoreactivating light suppressed the induction of SBCs by approximately 75%. Pre-UVR exposure to photoreactivating light had no effect on the induction of SBCs. Induction of hyperplasia also was suppressed to a similar extent by post-UVR photoreactivation treatment. These studies identify pyrimidine dimers in DNA as the major photoproduct involved in the induction of SBCs and hyperplasia in M. domestica by UVR

  19. Molecular characterization of monoclonal antibodies that inhibit acetylcholinesterase by targeting the peripheral site and backdoor region.

    Directory of Open Access Journals (Sweden)

    Yves Bourne

    Full Text Available The inhibition properties and target sites of monoclonal antibodies (mAbs Elec403, Elec408 and Elec410, generated against Electrophorus electricus acetylcholinesterase (AChE, have been defined previously using biochemical and mutagenesis approaches. Elec403 and Elec410, which bind competitively with each other and with the peptidic toxin inhibitor fasciculin, are directed toward distinctive albeit overlapping epitopes located at the AChE peripheral anionic site, which surrounds the entrance of the active site gorge. Elec408, which is not competitive with the other two mAbs nor fasciculin, targets a second epitope located in the backdoor region, distant from the gorge entrance. To characterize the molecular determinants dictating their binding site specificity, we cloned and sequenced the mAbs; generated antigen-binding fragments (Fab retaining the parental inhibition properties; and explored their structure-function relationships using complementary x-ray crystallography, homology modeling and flexible docking approaches. Hypermutation of one Elec403 complementarity-determining region suggests occurrence of antigen-driven selection towards recognition of the AChE peripheral site. Comparative analysis of the 1.9Å-resolution structure of Fab408 and of theoretical models of its Fab403 and Fab410 congeners evidences distinctive surface topographies and anisotropic repartitions of charges, consistent with their respective target sites and inhibition properties. Finally, a validated, data-driven docking model of the Fab403-AChE complex suggests a mode of binding at the PAS that fully correlates with the functional data. This comprehensive study documents the molecular peculiarities of Fab403 and Fab410, as the largest peptidic inhibitors directed towards the peripheral site, and those of Fab408, as the first inhibitor directed toward the backdoor region of an AChE and a unique template for the design of new, specific modulators of AChE catalysis.

  20. Acetylcholinesterase activity, cohabitation with floricultural workers, and blood pressure in Ecuadorian children.

    Science.gov (United States)

    Suarez-Lopez, Jose R; Jacobs, David R; Himes, John H; Alexander, Bruce H

    2013-05-01

    Acetylcholinesterase (AChE) inhibitors are commonly used pesticides that can effect hemodynamic changes through increased cholinergic stimulation. Children of agricultural workers are likely to have paraoccupational exposures to pesticides, but the potential physiological impact of such exposures is unclear. We investigated whether secondary pesticide exposures were associated with blood pressure and heart rate among children living in agricultural Ecuadorian communities. This cross-sectional study included 271 children 4-9 years of age [51% cohabited with one or more flower plantation workers (mean duration, 5.2 years)]. Erythrocyte AChE activity was measured using the EQM Test-mate system. Linear regression models were used to estimate associations of systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate with AChE activity, living with flower workers, duration of cohabitation with a flower worker, number of flower workers in the child's home, and number of practices that might increase children's exposure to pesticides. Mean (± SD) AChE activity was 3.14 ± 0.49 U/mL. A 1-U/mL decrease in AChE activity was associated with a 2.86-mmHg decrease in SBP (95% CI: -5.20, -0.53) and a 2.89-mmHg decrease in DBP (95% CI: -5.00, -0.78), after adjustment for potential confounders. Children living with flower workers had lower SBP (-1.72 mmHg; 95% CI: -3.53, 0.08) than other children, and practices that might increase exposure also were associated with lower SBP. No significant associations were found between exposures and heart rate. Our findings suggest that subclinical secondary exposures to pesticides may affect vascular reactivity in children. Additional research is needed to confirm these findings.

  1. Antioxidant and acetylcholinesterase inhibitory activities of ginger root (Zingiber officinale Roscoe) extract.

    Science.gov (United States)

    Tung, Bui Thanh; Thu, Dang Kim; Thu, Nguyen Thi Kim; Hai, Nguyen Thanh

    2017-05-04

    Background Zingiber officinale Roscoe has been used in traditional medicine for the treatment of neurological disorder. This study aimed to investigate the phenolic contents, antioxidant, acetylcholinesterase enzyme (AChE) inhibitory activities of different fraction of Z. officinale root grown in Vietnam. Methods The roots of Z. officinale are extracted with ethanol 96 % and fractionated with n-hexane, ethyl acetate (EtOAc) and butanol (BuOH) solvents. These fractions evaluated the antioxidant activity by 1,1-Diphenyl -2-picrylhydrazyl (DPPH) assay and AChE inhibitory activity by Ellman's colorimetric method. Results Our data showed that the total phenolic content of EtOAc fraction was highest equivalents to 35.2±1.4 mg quercetin/g of fraction. Our data also demonstrated that EtOAc fraction had the strongest antioxidant activity with IC50 was 8.89±1.37 µg/mL and AChE inhibitory activity with an IC50 value of 22.85±2.37 μg/mL in a dose-dependent manner, followed by BuOH fraction and the n-hexane fraction is the weakest. Detailed kinetic analysis indicated that EtOAc fraction was mixed inhibition type with Ki (representing the affinity of the enzyme and inhibitor) was 30.61±1.43 µg/mL. Conclusions Our results suggest that the EtOAc fraction of Z. officinale may be a promising source of AChE inhibitors for Alzheimer's disease.

  2. Protection from the toxicity of diisopropylfluorophosphate by adeno-associated virus expressing acetylcholinesterase

    International Nuclear Information System (INIS)

    Li Bin; Duysen, Ellen G.; Poluektova, Larisa Y.; Murrin, L. Charles; Lockridge, Oksana

    2006-01-01

    Organophosphorus esters (OP) are highly toxic chemicals used as pesticides and nerve agents. Their acute toxicity is attributed to inhibition of acetylcholinesterase (AChE, EC 3.1.1.7) in nerve synapses. Our goal was to find a new therapeutic for protection against OP toxicity. We used a gene therapy vector, adeno-associated virus serotype 2 (AAV-2), to deliver murine AChE to AChE-/- mice that have no endogenous AChE activity. The vector encoded the most abundant form of AChE: exons 2, 3, 4, and 6. Two-day old animals, with an immature immune system, were injected. AChE delivered intravenously was expressed up to 5 months in plasma, liver, heart, and lung, at 5-15% of the level in untreated wild-type mice. A few mice formed antibodies, but antibodies did not block AChE activity. The plasma AChE was a mixture of dimers and tetramers. AChE delivered intramuscularly had 40-fold higher activity levels than in wild-type muscle. None of the AChE was collagen-tailed. No retrograde transport through the motor neurons to the central nervous system was detected. AChE delivered intrastriatally assembled into tetramers. In brain, the AAV-2 vector transduced neurons, but not astrocytes and microglia. Vector-treated AChE-/- mice lived longer than saline-treated controls. AChE-/- mice were protected from diisopropylfluorophosphate-induced respiratory failure when the vector was delivered intravenously, but not intrastriatally. Since vector-treated animals had no AChE activity in diaphragm muscle, protection from respiratory failure came from AChE in other tissues. We conclude that AChE scavenged OP and in this way protected the activity of butyrylcholinesterase (BChE, EC 3.1.1.8) in motor endplates

  3. Drosophila acetylcholinesterase: demonstration of a glycoinositol phospholipid anchor and an endogenous proteolytic cleavage

    International Nuclear Information System (INIS)

    Haas, R.; Marshall, T.L.; Rosenberry, T.L.

    1988-01-01

    The presence of a glycoinositol phospholipid anchor Drosophila acetylcholinesterase (AChE) was shown by several criteria. Chemical analysis of highly purified Drosophila AChE demonstrated approximately one residue of inositol per enzyme subunit. Selective cleavage by Staphylococcus aureus phosphatidylinositol-specific phospholipase C (PI-PLC) was tested with Drosophila AChE radiolabeled by the photoactivatable affinity probe 3-(trifluoromethyl)-3-(m-[ 125 I]iodophenyl)diazirine ([ 125 I]TID), a reagent that specifically labels the lipid moiety of glycoinositol phospholipid-anchored proteins. Digestion with PI-PLC released 75% of this radiolabel from the protein. Gel electrophoresis of Drosophila AChE in sodium dodecyl sulfate indicated prominent 55- and 16-kDa bands and a faint 70-kDa band. The [ 125 ]I]TID label was localized on the 55-kDa fragment, suggesting that this fragment is the C-terminal portion of the protein. In support of this conclusion, a sensitive microsequencing procedure that involved manual Edman degradation combined with radiomethylation was used to determine residues 2-5 of the 16-kDa fragment. Comparison with the Drosophila AChE cDNA sequence confirmed that the 16-kDa fragment includes the N-terminus of AChE. Furthermore, the position of the N-terminal amino acid of the mature Drosophila AChE is closely homologous to that of Torpedo AChE. The presence of radiomethylatable ethanolamine in both 16- and 55-kDa fragments was also confirmed. Thus, Drosophila AChE may include a second posttranslational modification involving ethanolamine

  4. Chlorpyrifos and chlorpyrifos-oxon inhibit axonal growth by interfering with the morphogenic activity of acetylcholinesterase

    International Nuclear Information System (INIS)

    Yang Dongren; Howard, Angela; Bruun, Donald; Ajua-Alemanj, Mispa; Pickart, Cecile; Lein, Pamela J.

    2008-01-01

    A primary role of acetylcholinesterase (AChE) is regulation of cholinergic neurotransmission by hydrolysis of synaptic acetylcholine. In the developing nervous system, however, AChE also functions as a morphogenic factor to promote axonal growth. This raises the question of whether organophosphorus pesticides (OPs) that are known to selectively bind to and inactivate the enzymatic function of AChE also interfere with its morphogenic function to perturb axonogenesis. To test this hypothesis, we exposed primary cultures of sensory neurons derived from embryonic rat dorsal root ganglia (DRG) to chlorpyrifos (CPF) or its oxon metabolite (CPFO). Both OPs significantly decreased axonal length at concentrations that had no effect on cell viability, protein synthesis or the enzymatic activity of AChE. Comparative analyses of the effects of CPF and CPFO on axonal growth in DRG neurons cultured from AChE nullizygous (AChE -/- ) versus wild type (AChE +/+ ) mice indicated that while these OPs inhibited axonal growth in AChE +/+ DRG neurons, they had no effect on axonal growth in AChE -/- DRG neurons. However, transfection of AChE -/- DRG neurons with cDNA encoding full-length AChE restored the wild type response to the axon inhibitory effects of OPs. These data indicate that inhibition of axonal growth by OPs requires AChE, but the mechanism involves inhibition of the morphogenic rather than enzymatic activity of AChE. These findings suggest a novel mechanism for explaining not only the functional deficits observed in children and animals following developmental exposure to OPs, but also the increased vulnerability of the developing nervous system to OPs

  5. Hyperglycemia induces memory impairment linked to increased acetylcholinesterase activity in zebrafish (Danio rerio).

    Science.gov (United States)

    Capiotti, Katiucia Marques; De Moraes, Daiani Almeida; Menezes, Fabiano Peres; Kist, Luiza Wilges; Bogo, Maurício Reis; Da Silva, Rosane Souza

    2014-11-01

    Diabetes mellitus, which causes hyperglycemia, affects the central nervous system and can impairs cognitive functions, such as memory. The aim of this study was to investigate the effects of hyperglycemia on memory as well as on the activity of acethylcholinesterase. Hyperglycemia was induced in adult zebrafish by immersion in glucose 111mM by 14 days. The animals were divided in 4 groups: control, glucose-treated, glucose-washout 7-days and glucose-washout 14-days. We evaluated the performance in inhibitory avoidance task and locomotor activity. We also determined acethylcholinesterase activity and gene expression from whole brain. In order to counteract the effect of hyperglycemia underlined by effects on acethylcholinesterase activity, we treated the animals with galantamine (0.05ng/g), an inhibitor of this enzyme. Also we evaluated the gene expression of insulin receptor and glucose transporter from zebrafish brain. The hyperglycemia promoted memory deficit in adult zebrafish, which can be explained by increased AChE activity. The ache mRNA levels from zebrafish brain were decrease in 111mM glucose group and returned to normal levels after 7 days of glucose withdrawal. Insulin receptors (insra-1, insra-2, insrb-1 and insrb-2) and glut-3 mRNA levels were not significantly changed. Our results also demonstrated that galantamine was able to reverse the memory deficit caused by hyperglycemia, demonstrating that these effects involve modulation of AChE activity. These data suggest that the memory impairment induced by hyperglycemia is underlined by the cholinergic dysfunction caused by the mechanisms involving the control of acetylcholinesterase function and gene expression. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Anti-acetylcholinesterase activity and antioxidant properties of extracts and fractions of Carpolobia lutea.

    Science.gov (United States)

    Nwidu, Lucky Legbosi; Elmorsy, Ekramy; Thornton, Jack; Wijamunige, Buddhika; Wijesekara, Anusha; Tarbox, Rebecca; Warren, Averil; Carter, Wayne Grant

    2017-12-01

    There is an unmet need to discover new treatments for Alzheimer's disease. This study determined the anti-acetylcholinesterase (AChE) activity, DPPH free radical scavenging and antioxidant properties of Carpolobia lutea G. Don (Polygalaceae). The objective of this study is to quantify C. lutea anti-AChE, DPPH free radical scavenging, and antioxidant activities and cell cytotoxicity. Plant stem, leaves and roots were subjected to sequential solvent extractions, and screened for anti-AChE activity across a concentration range of 0.02-200 μg/mL. Plant DPPH radical scavenging activity, reducing power, and total phenolic and flavonoid contents were determined, and cytotoxicity evaluated using human hepatocytes. Carpolobia lutea exhibited concentration-dependent anti-AChE activity. The most potent inhibitory activity for the stem was the crude ethanol extract and hexane stem fraction oil (IC 50  = 140 μg/mL); for the leaves, the chloroform leaf fraction (IC 50  = 60 μg/mL); and for roots, the methanol, ethyl acetate and aqueous root fractions (IC 50  = 0.3-3 μg/mL). Dose-dependent free radical scavenging activity and reducing power were observed with increasing stem, leaf or root concentration. Total phenolic contents were the highest in the stem: ∼632 mg gallic acid equivalents/g for a hexane stem fraction oil. Total flavonoid content was the highest in the leaves: ∼297 mg quercetin equivalents/g for a chloroform leaf fraction. At 1 μg/mL, only the crude ethanol extract oil was significantly cytotoxic to hepatocytes. Carpolobia lutea possesses anti-AChE activity and beneficial antioxidant capacity indicative of its potential development as a treatment of Alzheimer's and other diseases characterized by a cholinergic deficit.

  7. Effects of carbofuran, diuron, and nicosulfuron on acetylcholinesterase activity in goldfish (Carassius auratus).

    Science.gov (United States)

    Bretaud, S; Toutant, J P; Saglio, P

    2000-10-01

    Juvenile goldfish (Carassius auratus) were exposed to three widely used pesticides; carbofuran, diuron, and nicosulfuron. Acetylcholinesterase (AChE) activity and molecular forms of AChE were first characterized in brain and skeletal muscle of unexposed fish. Skeletal muscle had higher AChE activity than brain (306 and 215 nmol/min/mg protein, respectively). In brain, four molecular forms of AChE were found: A12, G4, G2, and G1. In the muscle, three molecular forms were found A12, A8, and G2. AChE activity was then evaluated in both tissues of fish exposed to different concentration of pesticides (5, 50, and 500 microg/L) for 6, 12, 24, and 48 h. In brain, AChE activity was significantly inhibited during all the periods of exposure in response to 50 microg/L (19-28%) and 500 microg/L (85-87%) carbofuran. Such effect was observed in the muscle only at 500 microg/L (86-92%). Carbofuran had no effect on the distribution of molecular forms. Significant inhibitions (9-12%) of brain AChE activity were also observed in response to diuron and nicosulfuron at 500 microg/L during all periods of exposure and for 50 microg/L nicosulfuron after 24 and 48 h. This study pointed out short-term effects of exposure to sublethal concentrations of the three pesticides, ranging among different chemical families, on brain and muscle AChE in goldfish. Copyright 2000 Academic Press.

  8. Acetylcholinesterase inhibition and locomotor function after motor-sensory cortex impact injury.

    Science.gov (United States)

    Holschneider, Daniel P; Guo, Yumei; Roch, Margareth; Norman, Keith M; Scremin, Oscar U

    2011-09-01

    Traumatic brain injury (TBI) induces transient or persistent dysfunction of gait and balance. Enhancement of cholinergic transmission has been reported to accelerate recovery of cognitive function after TBI, but the effects of this intervention on locomotor activity remain largely unexplored. The hypothesis that enhancement of cholinergic function by inhibition of acetylcholinesterase (AChE) improves locomotion following TBI was tested in Sprague-Dawley male rats after a unilateral controlled cortical impact (CCI) injury of the motor-sensory cortex. Locomotion was tested by time to fall on the constant speed and accelerating Rotarod, placement errors and time to cross while walking through a horizontal ladder, activity monitoring in the home cages, and rearing behavior. Assessments were performed the 1st and 2nd day and the 1st, 2nd, and 3rd week after TBI. The AChE inhibitor physostigmine hemisulfate (PHY) was administered continuously via osmotic minipumps implanted subcutaneously at the rates of 1.6-12.8 μmol/kg/day. All measures of locomotion were impaired by TBI and recovered to initial levels between 1 and 3 weeks post-TBI, with the exception of the maximum speed achievable on the accelerating Rotarod, as well as rearing in the open field. PHY improved performance in the accelerating Rotarod at 1.6 and 3.2 μmol/kg/day (AChE activity 95 and 78% of control, respectively), however, higher doses induced progressive deterioration. No effect or worsening of outcomes was observed at all PHY doses for home cage activity, rearing, and horizontal ladder walking. Potential benefits of cholinesterase inhibition on locomotor function have to be weighed against the evidence of the narrow range of useful doses.

  9. Targeting acetylcholinesterase: identification of chemical leads by high throughput screening, structure determination and molecular modeling.

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    Lotta Berg

    Full Text Available Acetylcholinesterase (AChE is an essential enzyme that terminates cholinergic transmission by rapid hydrolysis of the neurotransmitter acetylcholine. Compounds inhibiting this enzyme can be used (inter alia to treat cholinergic deficiencies (e.g. in Alzheimer's disease, but may also act as dangerous toxins (e.g. nerve agents such as sarin. Treatment of nerve agent poisoning involves use of antidotes, small molecules capable of reactivating AChE. We have screened a collection of organic molecules to assess their ability to inhibit the enzymatic activity of AChE, aiming to find lead compounds for further optimization leading to drugs with increased efficacy and/or decreased side effects. 124 inhibitors were discovered, with considerable chemical diversity regarding size, polarity, flexibility and charge distribution. An extensive structure determination campaign resulted in a set of crystal structures of protein-ligand complexes. Overall, the ligands have substantial interactions with the peripheral anionic site of AChE, and the majority form additional interactions with the catalytic site (CAS. Reproduction of the bioactive conformation of six of the ligands using molecular docking simulations required modification of the default parameter settings of the docking software. The results show that docking-assisted structure-based design of AChE inhibitors is challenging and requires crystallographic support to obtain reliable results, at least with currently available software. The complex formed between C5685 and Mus musculus AChE (C5685•mAChE is a representative structure for the general binding mode of the determined structures. The CAS binding part of C5685 could not be structurally determined due to a disordered electron density map and the developed docking protocol was used to predict the binding modes of this part of the molecule. We believe that chemical modifications of our discovered inhibitors, biochemical and biophysical

  10. Effect of ions on the activity of brain acetylcholinesterase from tropical fish

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    Caio Rodrigo Dias Assis

    2015-07-01

    Full Text Available Objective: To investigate the effect of ions on brain acetylcholinesterase (AChE; EC 3.1.1.7 activities from economic important fish [pirarucu, Arapaima gigas; tambaqui, Colossoma macropomum; cobia, Rachycentron canadum (R. canadum and Nile tilapia, Oreochromis niloticus (O. niloticus] comparing with a commercial enzyme from electric eel [Electrophorus electricus (E. electricus]. Methods: The in vitro exposure was performed at concentrations ranging from 0.001 to 10 mmol/L (except for ethylene diamine tetraacetic acid; up to 150 mmol/L. Inhibition kinetics on R. canadum and O. niloticus were also observed through four methods (Michaelis-Menten, Lineweaver-Burk, Dixon and Cornish-Bowden plots in order to investigate the type of inhibition produced by some ions. Results: Hg 2+ , As 3+ , Cu 2+ , Zn 2+ , Cd 2+ caused inhibition in all the species under study. Ca 2+ , Mg 2+ and Mn 2+ induced slight activation in R. canadum enzyme while Pb 2+ , Ba 2+ , Fe 2+ , Li + inhibited the AChE from some of the analyzed species. The lowest IC 50 and Ki values were estimated for E. electricus AChE in presence of Hg 2+ , Pb 2+ , Zn 2+ . Under our experimental conditions, the results for R. canadum and O. niloticus, As 3+ , Cu 2+ , Cd 2+ , Pb 2+ and Zn 2+ showed a non- competitive/mixed-type inhibition, while Hg 2+ inhibited the enzyme in a mixed/competitive- like manner. Conclusions: E. electricus AChE activity was affected by ten of fifteen ions under study showing that this enzyme could undergo interference by these ions when used as pesticide biosensor in environmental analysis. This hindrance would be less relevant for the crude extracts.

  11. Acetylcholinesterase and butyrylcholinesterase inhibitory activity of some selected Nigerian medicinal plants

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    Taiwo O. Elufioye

    Full Text Available Plants have been found to be useful as memory enhansers as well as antiaging. Twenty two of such plants from sixteen families were investigated for their acetylcholinesterase (AChE and butyrylcholinesterase (BuChE inhibitory activities using the in vitro Ellman's spectrophotometric and in situ bioautographic methods with physostigmine as standard. At least three morphological parts were examined for each of the plants investigated and the test concentration was 42.5 µg/ mL. Some plants were active on both enzymes though with some morphological parts being more active than others. The root bark of Spondias mombin showed the highest activity to the two enzymes; 64.77% and 83.94% on AChE and BuChE respectively. Other plant parts of the selected plants exhibited some remarkable selectivity in their actions. Those selectively active against AChE were Alchornia laxiflora stem bark (41.12% and root bark, Callophyllum inophyllurn root bark (56.52%. The leaves of C. jagus (74.25%, Morinda lucida leaves (40.15%, Peltophorum pterocarpum leaves and stem bark (49.5% and 68.85%, respectively, physiostigmine gave 90.31% inhibition. Generally higher activities were found against BuChE. Bombax bromoposenze leaves, root bark and stem bark were particularly active. The inhibition was over 80%. Other selective plant parts are the leaves Antiaris africana, Cissampelos owarensis aerial parts (78.96%, Combretum molle leaves and stem bark (90.42% and 88.13%, respectively, Dioscorea dumentorum root bark and tuber (over 87%, G. kola leaves, Markhamia tomentosa root bark, Pycnanthus angolensis stem bark and Tetrapleura tetraptera leaves. Most of these plants are taken as food or are food ingredients in Nigeria and may account for the low incidence of Alzheimer's disease in the country and may play certain roles in the mediation of the disease.

  12. Immobilization of acetylcholinesterase via biocompatible interface of silk fibroin for detection of organophosphate and carbamate pesticides

    Energy Technology Data Exchange (ETDEWEB)

    Xue Rui [College of Environmental and Energy Engineering, Beijing University of Technology, Beijing 100124 (China); Kang Tianfang, E-mail: kangtf@yahoo.cn [College of Environmental and Energy Engineering, Beijing University of Technology, Beijing 100124 (China); Lu Liping; Cheng Shuiyuan [College of Environmental and Energy Engineering, Beijing University of Technology, Beijing 100124 (China)

    2012-06-01

    An amperometric biosensor for the detection of organophosphate and carbamate pesticides was developed based on the immobilization of acetylcholinesterase (AChE) on regenerated silk fibroin (SF) matrix by non-covalent adsorption. SF and AChE were coated sequentially on the surface of the glassy carbon electrode (GCE) which was modified with multiwall carbon nanotube (MWNTs). The obtained biosensor was denoted as AChE-SF/MWNTs/GCE. The atomic force microscopy images showed that the SF matrix provided a more homogeneous interface for the AChE immobilization. The aggregation of immobilizing AChE was therefore avoided. The cyclic voltammogram of thiocholine at this biosensor exhibited a well defined oxidation peak at 0.667 V (vs. SCE). The inhibition rate of methyl parathion to the immobilized AChE was proportional to the logarithm of the concentration of methyl parathion over the range of the concentration of methyl parathion from 3.5 Multiplication-Sign 10{sup -6} to 2.0 Multiplication-Sign 10{sup -3} M with a detection limit of 5.0 Multiplication-Sign 10{sup -7} M. Similarly, the linearly response range of carbaryl was from 1.0 Multiplication-Sign 10{sup -7} to 3.0 Multiplication-Sign 10{sup -5} M with a detection limit of 6.0 Multiplication-Sign 10{sup -8} M. The experimental results indicate that AChE not only can be immobilized steadily on the SF matrix, but also the bioactivity of immobilizing AChE can be preserved effectively.

  13. Calretinin immunohistochemistry versus improvised rapid Acetylcholinesterase histochemistry in the evaluation of colorectal biopsies for Hirschsprung disease

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    Lokendra Yadav

    2014-01-01

    Full Text Available Background: Acetylcholinesterase (AChE histochemistry on rectal mucosal biopsies accurately diagnoses Hirschsprung disease (HD, but is not widely employed as it requires special tissue handling and pathologist expertise. Calretinin immunohistochemistry (IHC has been reported to be comparable to AChE staining with the loss of expression correlating with aganglionosis. Aim: The aim was to evaluate calretinin IHC as a primary diagnostic tool in comparison to the improvised rapid AChE technique in the diagnosis of HD. Materials and Methods: A total of 74 rectal biopsies (18 fresh frozen - 18 cases, 56 formalin fixed - 33 cases from 51 cases of suspect HD were evaluated with hematoxylin and eosin/AChE/Calretinin. Ten biopsies each from ganglionated and aganglionated segments served as positive and negative controls. Ileal (3, appendiceal (3 and ring bowel (2 biopsies were also included. Two pathologists blinded to the clinical details evaluated the histomorphology with AChE and calretinin. Observations were statistically analyzed and Cohen′s k coefficient employed to assess agreement between two pathologists and calretinin and the AChE. Results: The study confirmed HD in 26 and non-HD in 25 cases. There were 7 neonates, 5 low level biopsies and 14 "inadequate" biopsies. The results of calretinin were comparable with AChE with a statistically significant measure of agreement of k = 0.973 between the two. One false-positive case of HD was noted with calretinin. The advantages and disadvantages of calretinin versus AChE are discussed. Conclusion: Calretinin is a reliable single immune marker for ruling out HD by its specific positive mucosal staining of formalin fixed rectal biopsy. The improvised AChE staining remains indispensable to confirm HD on fresh biopsies and thus, along with calretinin IHC maximizes the diagnostic accuracy of HD in difficult cases.

  14. Acetylcholinesterase Regulates Skeletal In Ovo Development of Chicken Limbs by ACh-Dependent and -Independent Mechanisms.

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    Janine Spieker

    Full Text Available Formation of the vertebrate limb presents an excellent model to analyze a non-neuronal cholinergic system (NNCS. Here, we first analyzed the expression of acetylcholinesterase (AChE by IHC and of choline acetyltransferase (ChAT by ISH in developing embryonic chicken limbs (stages HH17-37. AChE outlined formation of bones, being strongest at their distal tips, and later also marked areas of cell death. At onset, AChE and ChAT were elevated in two organizing centers of the limb anlage, the apical ectodermal ridge (AER and zone of polarizing activity (ZPA, respectively. Thereby ChAT was expressed shortly after AChE, thus strongly supporting a leading role of AChE in limb formation. Then, we conducted loss-of-function studies via unilateral implantation of beads into chicken limb anlagen, which were soaked in cholinergic components. After varying periods, the formation of cartilage matrix and of mineralizing bones was followed by Alcian blue (AB and Alizarin red (AR stainings, respectively. Both acetylcholine (ACh- and ChAT-soaked beads accelerated bone formation in ovo. Notably, inhibition of AChE by BW284c51, or by the monoclonal antibody MAB304 delayed cartilage formation. Since bead inhibition of BChE was mostly ineffective, an ACh-independent action during BW284c51 and MAB304 inhibition was indicated, which possibly could be due to an enzymatic side activity of AChE. In conclusion, skeletogenesis in chick is regulated by an ACh-dependent cholinergic system, but to some extent also by an ACh-independent aspect of the AChE protein.

  15. Molecular cloning and characterization of an acetylcholinesterase cDNA in the brown planthopper, Nilaparvata lugens.

    Science.gov (United States)

    Yang, Zhifan; Chen, Jun; Chen, Yongqin; Jiang, Sijing

    2010-01-01

    A full cDNA encoding an acetylcholinesterase (AChE, EC 3.1.1.7) was cloned and characterized from the brown planthopper, Nilaparvata lugens Stål (Hemiptera: Delphacidae). The complete cDNA (2467 bp) contains a 1938-bp open reading frame encoding 646 amino acid residues. The amino acid sequence of the AChE deduced from the cDNA consists of 30 residues for a putative signal peptide and 616 residues for the mature protein with a predicted molecular weight of 69,418. The three residues (Ser242, Glu371, and His485) that putatively form the catalytic triad and the six Cys that form intra-subunit disulfide bonds are completely conserved, and 10 out of the 14 aromatic residues lining the active site gorge of the AChE are also conserved. Northern blot analysis of poly(A)+ RNA showed an approximately 2.6-kb transcript, and Southern blot analysis revealed there likely was just a single copy of this gene in N. lugens. The deduced protein sequence is most similar to AChE of Nephotettix cincticeps with 83% amino acid identity. Phylogenetic analysis constructed with 45 AChEs from 30 species showed that the deduced N. lugens AChE formed a cluster with the other 8 insect AChE2s. Additionally, the hypervariable region and amino acids specific to insect AChE2 also existed in the AChE of N. lugens. The results revealed that the AChE cDNA cloned in this work belongs to insect AChE2 subgroup, which is orthologous to Drosophila AChE. Comparison of the AChEs between the susceptible and resistant strains revealed a point mutation, Gly185Ser, is likely responsible for the insensitivity of the AChE to methamidopho in the resistant strain.

  16. Acotiamide hydrochloride (Z-338) enhances gastric motility and emptying by inhibiting acetylcholinesterase activity in rats.

    Science.gov (United States)

    Kawachi, Masanao; Matsunaga, Yugo; Tanaka, Takao; Hori, Yuko; Ito, Katsunori; Nagahama, Kenji; Ozaki, Tomoko; Inoue, Naonori; Toda, Ryoko; Yoshii, Kazuyoshi; Hirayama, Masamichi; Kawabata, Yoshihiro; Takei, Mineo

    2011-09-01

    In clinical trials, acotiamide hydrochloride (acotiamide: Z-338) has been reported to be useful in the treatment of functional dyspepsia. Here, we investigated the effects of acotiamide on gastric contraction and emptying activities in rats in comparison with itopride hydrochloride (itopride) and mosapride citrate (mosapride). We also examined in vitro the compound's inhibitory effect on acetylcholinesterase (AChE) activity derived from rat stomach. In in vivo studies, acotiamide (30 and 100mg/kg s.c.) and itopride (100mg/kg s.c.) markedly enhanced normal gastric antral motility in rats. In gastric motility dysfunction models, acotiamide (100mg/kg s.c.) and itopride (100mg/kg s.c.) improved both gastric antral hypomotility and the delayed gastric emptying induced by clonidine, an α(2)-adrenoceptor agonist. In contrast, mosapride (10mg/kg s.c.) had no effect on these models. Like the AChE inhibitors itopride (30 mg/kg s.c.) and neostigmine (10 μg/kg s.c.), acotiamide (10mg/kg s.c.) also clearly enhanced gastric body contractions induced by electrical stimulation of the vagus, which were abolished by atropine and hexamethonium, whereas mosapride (3 and 10mg/kg s.c.) did not. In in vitro studies, acotiamide concentration-dependently inhibited rat stomach-derived AChE activity (IC(50)=2.3 μmol/l). In addition, stomach tissue concentrations of acotiamide after administration at 10mg/kg s.c. were sufficient to produce inhibition of AChE activity in rat stomach. These results suggest that acotiamide stimulates gastric motility and improves gastric motility dysfunction in rats by inhibiting AChE activity, and may suggest a role for acotiamide in improving gastric motility dysfunction in patients with functional dyspepsia. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Reactivation of organophosphate-inhibited human acetylcholinesterase by isonitrosoacetone (MINA): a kinetic analysis.

    Science.gov (United States)

    Worek, Franz; Thiermann, Horst

    2011-11-15

    Treatment of poisoning by highly toxic organophosphorus compounds (OP) with atropine and an acetylcholinesterase (AChE) reactivator (oxime) is of limited effectiveness in case of different nerve agents and pesticides. One challenge is the reactivation of OP-inhibited brain AChE which shows inadequate success with charged pyridinium oximes. Recent studies with high doses of the tertiary oxime isonitrosoacetone (MINA) indicated a beneficial effect on central and peripheral AChE and on survival in nerve agent poisoned guinea pigs. Now, an in vitro study was performed to determine the reactivation kinetics of MINA with tabun-, sarin-, cyclosarin-, VX- and paraoxon-inhibited human AChE. MINA showed an exceptionally low affinity to inhibited AChE but, with the exception of tabun-inhibited AChE, a moderate to high reactivity. In comparison to the pyridinium oximes obidoxime, 2-PAM and HI-6 the affinity and reactivity of MINA was in most cases lower and in relation to the most effective reactivators, the second order reactivation constant of MINA was 500 to 3400-fold lower. Hence, high in vivo MINA concentrations would be necessary to achieve at least partial reactivation. This assumption corresponds to in vivo data showing a dose-dependent effect on reactivation and survival in animals. In view, of the toxic potential of MINA in animals human studies would be necessary to determine the tolerability and pharmacokinetics of MINA in order to enable a proper assessment of the value of this oxime as an antidote in OP poisoning. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  18. Natural monomeric form of fetal bovine serum acetylcholinesterase lacks the C-terminal tetramerization domain.

    Science.gov (United States)

    Saxena, Ashima; Hur, Regina S; Luo, Chunyuan; Doctor, Bhupendra P

    2003-12-30

    Acetylcholinesterase isolated from fetal bovine serum (FBS AChE) was previously characterized as a globular tetrameric form. Analysis of purified preparations of FBS AChE by gel permeation chromatography revealed the presence of a stable, catalytically active, monomeric form of this enzyme. The two forms could be distinguished from each other based on their molecular weight, hydrodynamic properties, kinetic properties, thermal stability, and the type of glycans they carry. No differences between the two forms were observed for the binding of classical inhibitors such as edrophonium and propidium or inhibitors that are current or potential drugs for the treatment of Alzheimer's disease such as (-) huperzine A and E2020; tacrine inhibited the monomeric form 2-3-fold more potently than the tetrameric form. Sequencing of peptides obtained from an in-gel tryptic digest of the monomer and tetramer by tandem mass spectrometry indicated that the tetramer consists of 583 amino acid residues corresponding to the mature form of the enzyme, whereas the monomer consists of 543-547 amino acid residues. The subunit molecular weight of the protein component of the monomer (major species) was determined to be 59 414 Da and that of the tetramer as 64 239 Da. The N-terminal of the monomer and the tetramer was Glu, suggesting that the monomer is not a result of truncation at the N-terminal. The only differences detected were at the C-terminus. The tetramer yielded the expected C-terminus, CSDL, whereas the C-terminus of the monomer yielded a mixture of peptides, of which LLSATDTLD was the most abundant. These results suggest that monomeric FBS AChE is trimmed at the C-terminus, and the results are consistent with the involvement of C-terminal amino acids in the assembly of monomers into tetramers.

  19. Acetylcholinesterase Inhibition and Antioxidant Activity of N-trans-Caffeoyldopamine and N-trans-Feruloyldopamine

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    Muamer Dizdar

    2018-04-01

    Full Text Available Phenolic acids and their derivatives found in nature are well-known for their potential biological activity. In this study, two amides derived from trans-caffeic/ferulic acid and dopamine were synthesized and characterized by Fourier-transform infrared spectroscopy (FTIR, mass spectrometry, proton and carbon-13 nuclear magnetic resonance spectroscopy. The compounds were tested for the inhibition of acetylcholinesterase (AChE from Electrophorus electricus and for antioxidant activity by scavenging 2,2-diphenyl-1-pycrylhydrazyl free radical (DPPH• and 2,2′-azinobis(3-ethylbenzothiazoline-6-sulphonic acid radical cation (ABTS•+, reducing ferric ions, and ferrous ions chelation. N-trans-Feruloyldopamine displayed the highest inhibitory effect on AChE with half-maximal inhibitory concentration (IC50 values of 8.52 μM. In addition, an in silico study was done to determine the most favorable AChE cluster with the synthesized compounds. Further, these clusters were investigated for binding positions at the lowest free binding energy. Both synthesized hydroxycinnamates were found to be better antioxidants than the parent acids in in vitro tests applied. N-trans-Caffeoyldopamine showed the best antioxidant activity in the three tested methods—against non-biological stable free radicals IC50 5.95 μM for DPPH•, 0.24 μM for the ABTS•+ method, and for reducing power (ascorbic acid equivalent (AAE 822.45 μmol/mmol—while for chelation activity against Fe2+ ions N-trans-feruloyldopamine had slightly better antioxidant activity (IC50 3.17 mM.

  20. Kinetics of Huperzine A Dissociation from Acetylcholinesterase via Multiple Unbinding Pathways.

    Science.gov (United States)

    Rydzewski, Jakub; Jakubowski, Rafal; Nowak, Wieslaw; Grubmüller, Helmut

    2018-05-01

    The dissociation of huperzine A (hupA) from Torpedo californica acetylcholinesterase (TcAChE) was investigated by 4-microsecond unbiased and biased all-atom molecular dynamics (MD) simulations in explicit solvent. We performed our study using memetic sampling (MS) for the determination of reaction pathways (RPs), metadynamics to calculate free energy, and maximum-likelihood estimation (MLE) to recover kinetic rates from unbiased MD simulations. Our simulations suggest that the dissociation of hupA occurs mainly via two RPs: a front-door along the axis of the active-site gorge (pwf) and through a new transient side-door (pws), i.e., formed by the Omega-loop (residues 67--94 of TcAChE). Analysis of the inhibitor unbinding along the RPs suggests that pws is opened transiently after hupA and the Omega-loop reach a low free-energy transition state characterized by the orientation of the pyridone group of the inhibitor directed toward the Omega-loop plane. Unlike pws, pwf does not require large structural changes of TcAChE to be accessible. The estimated free energies and rates agree well with available experimental data. The dissociation rates along the unbinding pathways are similar, suggesting that the dissociation of hupA along pws is likely to be relevant. This indicates that perturbations to hupA-TcAChE interactions could potentially induce pathway hopping. In summary, out results characterize the slow-onset inhibition of TcAChE by hupA, which may provide the structural and energetic basis for the rational design of the next-generation slow-onset inhibitors with optimized pharmacokinetic properties for the treatment of Alzheimer's disease.

  1. Comparison of oxime reactivation and aging of nerve agent-inhibited monkey and human acetylcholinesterases.

    Science.gov (United States)

    Luo, Chunyuan; Tong, Min; Maxwell, Donald M; Saxena, Ashima

    2008-09-25

    Non-human primates are valuable animal models that are used for the evaluation of nerve agent toxicity as well as antidotes and results from animal experiments are extrapolated to humans. It has been demonstrated that the efficacy of an oxime primarily depends on its ability to reactivate nerve agent-inhibited acetylcholinesterase (AChE). If the in vitro oxime reactivation of nerve agent-inhibited animal AChE is similar to that of human AChE, it is likely that the results of an in vivo animal study will reliably extrapolate to humans. Therefore, the goal of this study was to compare the aging and reactivation of human and different monkey (Rhesus, Cynomolgus, and African Green) AChEs inhibited by GF, GD, and VR. The oximes examined include the traditional oxime 2-PAM, two H-oximes HI-6 and HLo-7, and the new candidate oxime MMB4. Results indicate that oxime reactivation of all three monkey AChEs was very similar to human AChE. The maximum difference in the second-order reactivation rate constant between human and three monkey AChEs or between AChEs from different monkey species was 5-fold. Aging rate constants of GF-, GD-, and VR-inhibited monkey AChEs were very similar to human AChE except for GF-inhibited monkey AChEs, which aged 2-3 times faster than the human enzyme. The results of this study suggest that all three monkey species are suitable animal models for nerve agent antidote evaluation since monkey AChEs possess similar biochemical/pharmacological properties to human AChE.

  2. Structural characterization of acetylcholinesterase 1 from the sand fly Lutzomyia longipalpis (Diptera: Psychodidae).

    Science.gov (United States)

    Coutinho-Abreu, I V; Balbino, V Q; Valenzuela, J G; Sonoda, I V; Ramalho-Ortigão, J M

    2007-07-01

    Acetylcholinesterase (AChE) plays a key role in cholinergic impulse transmission, and it is the target enzyme for organophosphorus and carbamate insecticides. Two genes, AceI and AceII, have been characterized from different insect species, and point mutations in either gene can lead to significant resistance to these classes of insecticides. In this report, we describe the partial characterization of the AceI gene from Lutzomyia longipalpis (Lutz & Neiva) (Diptera: Psychodidae), and we show that the possibility exists for the development of a resistant phenotype to organophosphates and carbamates in sand flies. Our results point to the presence of a single AceI gene in L. longipalpis (LlAce1) and that AChE activity is inhibited by organophosphorus at a concentration of 5 x 10(-5) M. Regarding insecticide resistance, analysis of the truncated LlAce1 cDNA suggests that a single missense mutation leading to a glycine-to-serine substitution at amino acid position 119 (G119S) may arise in L. longipalpis, similar to what has been detected in Anopheles gambiae s.s. Another missense mutation involved in resistant phenotypes, F331W, detected in Culex tritaeniorhynchus Giles, is less likely to occur in L. longipalpis, because it faces codon constraint in this sand fly species. Comparison of the three-dimensional structures of the deduced amino acid sequence of the truncated LLAChE1 with that of An. gambiae and Cx. tritaeniorhynchus also suggests that similar structural modifications due to the missense amino acid changes in the active site gorge are detected in all three insects.

  3. Acetylcholinesterase potentiates [3H]fluorowillardiine and [3H]AMPA binding to rat cortical membranes

    International Nuclear Information System (INIS)

    Olivera, S.; Rodriguez-Ithurralde, D.; Henley, J.M.

    1999-01-01

    In addition to its action at cholinergic synapses acetylcholinesterase (AChE) has been proposed to modulate neuronal activity by mechanisms unrelated to the hydrolysis of acetylcholine. We have investigated the effects of AChE on the binding of the specific AMPA receptor agonists (S)-[ 3 H]5-fluorowillardiine ([ 3 H]FW) and [ 3 H]AMPA to rat cortical membranes. Pretreatment of membranes with AChE causes a dose-dependent increase in the binding of both radiolabelled agonists with a maximal increase to ∼60% above control. This increase is completely blocked by the specific AChE inhibitors propidium, physostigmine, DFP and BW 284C51. AChE pretreatment had no effect on [ 3 H]kainate binding. [ 3 H]FW binding to membranes from young (15-day-old) rats is four orders of magnitude more sensitive to AChE modulation than membranes from adult rats (EC 50 values of 4x10 -5 and 0.1 unit/ml, respectively) although the total percentage increase in binding is similar. Furthermore, the AChE-induced potentiation of [ 3 H]FW binding is Ca 2+ - and temperature-dependent suggesting an enzymatic action for AChE in this system. Saturation binding experiments with [ 3 H]FW to adult membranes reveal high and low affinity binding sites and demonstrate that the main action of AChE is to increase the B max of both sites. These findings suggest that modulation of AMPA receptors could provide a molecular mechanism of action for the previously reported effects of AChE in synapse formation, synaptic plasticity and neurodegeneration. (Copyright (c) 1999 Elsevier Science B.V., Amsterdam. All rights reserved.)

  4. The distribution and impact of common copy-number variation in the genome of the domesticated apple, Malus x domestica Borkh.

    Science.gov (United States)

    Boocock, James; Chagné, David; Merriman, Tony R; Black, Michael A

    2015-10-23

    Copy number variation (CNV) is a common feature of eukaryotic genomes, and a growing body of evidence suggests that genes affected by CNV are enriched in processes that are associated with environmental responses. Here we use next generation sequence (NGS) data to detect copy-number variable regions (CNVRs) within the Malus x domestica genome, as well as to examine their distribution and impact. CNVRs were detected using NGS data derived from 30 accessions of M. x domestica analyzed using the read-depth method, as implemented in the CNVrd2 software. To improve the reliability of our results, we developed a quality control and analysis procedure that involved checking for organelle DNA, not repeat masking, and the determination of CNVR identity using a permutation testing procedure. Overall, we identified 876 CNVRs, which spanned 3.5 % of the apple genome. To verify that detected CNVRs were not artifacts, we analyzed the B- allele-frequencies (BAF) within a single nucleotide polymorphism (SNP) array dataset derived from a screening of 185 individual apple accessions and found the CNVRs were enriched for SNPs having aberrant BAFs (P apple scab. We present the first analysis and catalogue of CNVRs in the M. x domestica genome. The enrichment of the CNVRs with R gene models and their overlap with gene loci of agricultural significance draw attention to a form of unexplored genetic variation in apple. This research will underpin further investigation of the role that CNV plays within the apple genome.

  5. Acetylcholinesterase immobilized capillary reactors coupled to protein coated magnetic beads: A new tool for plant extract ligand screening

    OpenAIRE

    Vanzolini, Kenia Lourenço; Jiang, Zhengjin; Zhang, Xiaoqi; Vieira, Lucas Campos Curcino; Corrêa, Arlene Gonçalvez; Cardoso, Carmen Lucia; Cass, Quezia Bezerra; Moaddel, Ruin

    2013-01-01

    The use of immobilized capillary enzyme reactors (ICERs) and enzymes coated to magnetic beads ((NT or CT)-MB) for ligand screening has been adopted as a new technique of high throughput screening (HTS). In this work the selected target was the enzyme acetylcholinesterase (AChE), which acts on the central nervous system and is a validated target for the treatment of Alzheimer’s disease, as well as for new insecticides. A new approach for the screening of plant extracts was developed based on t...

  6. Acetylcholinesterase inhibition and antioxidant activity of essential oils from Schinus areira L. and Schinus longifolia (Lindl.) Speg.

    Science.gov (United States)

    Murray, Ana P; Gurovic, Maria S Vela; Rodriguez, Silvana A; Murray, María G; Ferrero, Adriana A

    2009-06-01

    The essential oils of Schinus areira L. and S. longifolia (Lindl.) Speg. (Anacardiaceae) have been studied for their in vitro anti-acetylcholinesterase and antioxidant activities. The chemical composition of the oils obtained by hydrodistillation was determined by GC-MS. Fruit and leaf oils of S. areira were analyzed separately. The essential oil from S. longifolia elicited marked enzymatic inhibition (IC50 = 20.0 +/- 1.0 microg/mL) and showed radical scavenger activity (IC50 = 25.2 +/- 2.4 microg/mL). The essential oil from S. areira leaves was more active than that of the fruits in both bioassays.

  7. Exploring the multifunctionality of thioflavin- and deferiprone-based molecules as acetylcholinesterase inhibitors for potential application in Alzheimer's disease.

    Science.gov (United States)

    Telpoukhovskaia, Maria A; Patrick, Brian O; Rodríguez-Rodríguez, Cristina; Orvig, Chris

    2013-04-05

    Alzheimer's disease (AD) is a devastating neurodegenerative disease that affects millions of people worldwide. With no prevention or cure available, this progressive disease has a significant impact on society - dementia patients and their caretakers, healthcare systems, and the economy. Previously, we have reported initial developments of multifunctional drug candidates for AD based on two scaffolds - thioflavin-T and deferiprone. Individually, these molecules have shown several favorable functionalities, including dissociation of toxic amyloid-β aggregates, antioxidant and/or metal chelating ability that can pacify reactive oxygen species, plaque targeting, and blood-brain barrier penetration. In this work, the two scaffolds are augmented with a new functionality - acetylcholinesterase inhibition. This functionality is incorporated by derivatization with a carbamate group, which is the active group in some AD drugs currently in the market. We present the rationale for designing three novel compounds, their synthesis and characterization, including X-ray crystallographic data, and encouraging results from in vitro and computational acetylcholinesterase inhibition studies. Also, we evaluate the compounds as potential drug candidates by Lipinski's rules and cytotoxicity studies in a neuronal cell line. Overall, we demonstrate the feasibility of improving on two well established scaffolds, as well as show in vitro efficacy plus initial mode of action and biological compatibility data.

  8. Anti-acetylcholinesterase and Antioxidant Activity of Essential Oils from Hedychium gardnerianum Sheppard ex Ker-Gawl

    Directory of Open Access Journals (Sweden)

    José Silvino Rosa

    2012-03-01

    Full Text Available Acetylcholinesterase inhibition, antioxidant and cytotoxic activities of Hedychium gardnerianum leaf essential oils from S. Miguel Island were determined. All the oils inhibited acetylcholinesterase, with IC50 values of approximately 1 mg/mL, showing no statistical differences between collection sites. Three oils presented mixed inhibition, whilst one was almost truly competitive. This activity can be attributed to the presence of sesquiterpenes, which constituted more than 60% of the composition of the oils. Regarding the antioxidant activity as measured by the DPPH method, all the oils presented activities similar to reference compounds, although with statistical differences between collection sites. Cytotoxicity measured using Artemia salina classified these oils as moderately toxic, with LC50 values ranging from 300 to 500 µg/mL. These results indicate a possible application of these oils in aromatherapy as coadjuvants in the treatment of cognitive diseases such as Alzheimer, since they may contribute to increase acetylcholine in cholinergic neurons and simultaneously fight deleterious oxidations responsible by neurological degeneration.

  9. Acetylcholinesterase inhibition and antibacterial activity of Mondia whitei adventitious roots and ex vitro-grown somatic embryogenic-biomass

    Directory of Open Access Journals (Sweden)

    Ponnusamy Baskaran

    2016-10-01

    Full Text Available Mondia whitei (Hook.f. Skeels is an important endangered medicinal and commercial plant in South Africa. In vitro propagation systems are required for biomass production and bioactivity analysis to supplement wild resources/stocks. Adventitious roots from somatic embryogenic explants using suspension culture and ex vitro-grown plants produced via somatic embryogenesis were established using different plant growth regulator treatments. The adventitious root biomass and different parts of ex vitro-grown and mother plants were used to investigate the potential for acetylcholinesterase (AChE and antibacterial activities. Adventitious roots derived from 2.5 µM indole-3-acetic acid (IAA treatments and ex vitro-grown plants derived from meta-topolin riboside (mTR and IAA treatments gave the best AChE and antibacterial activities. The in vitro-established M. whitei and ex vitro biomass have comparable ability to function as inhibitors of acetylcholinesterase and antibacterial agents, and can be used as potent bioresources in traditional medicine

  10. Highly Sensitive Detection of Organophosphate Insecticides Using Biosensors Based on Genetically Engineered Acetylcholinesterase and Poly(3,4-Ethylenedioxythiophene

    Directory of Open Access Journals (Sweden)

    Tomasz Sikora

    2011-01-01

    Full Text Available A poly(3,4-ethylenedioxythiophene (PEDOT conducting ink is presented as a new electroactive material to be incorporated in acetylcholinesterase-(AChE- based screen printed biosensors, acting not only as a conducting template but also as an electrochemical mediator for thiocholine oxidation. Two different strategies have been studied for the chemical synthesis of PEDOT: (a a classical oxidative polymerisation and (b a more innovative enzymatic polymerisation, giving a water-soluble PEDOT. The use of this water-soluble conducting polymer as mediator in screen-printed biosensors enables its deposition by printing like the rest of the layers. Highly sensitive acetylcholinesterase-(AChE- based screen-printed biosensors have been constructed using both classical and enzymatic PEDOT, in combination with genetically modified AChE. These electrodes allow the measurement of thiocholine oxidation at potentials of 100 mV versus Ag/AgCl reference electrode through the mediation of PEDOT. Inhibition of thiocholine production in presence of CPO allow for detection of this pesticide in concentrations as low as 1·10−10 M.

  11. Guarana (Paullinia cupana) ameliorates memory impairment and modulates acetylcholinesterase activity in Poloxamer-407-induced hyperlipidemia in rat brain.

    Science.gov (United States)

    Ruchel, Jader B; Braun, Josiane B S; Adefegha, Stephen A; Guedes Manzoni, Alessandra; Abdalla, Fátima H; de Oliveira, Juliana S; Trelles, Kelly; Signor, Cristiane; Lopes, Sônia T A; da Silva, Cássia B; Castilhos, Lívia G; Rubin, Maribel A; Leal, Daniela B R

    2017-01-01

    Hyperlipidemia is a risk factor for the development of cognitive dysfunction and atherosclerosis. Natural compounds have recently received special attention in relation to the treatment of disease due to their low cost and wide margin of safety. Thus, the aim of this study was to determine the possible preventive effect of guarana powder (Paullinia cupana) on memory impairment and acetylcholinesterase (AChE) activity in the brain structures of rats with Poloxamer-407-induced hyperlipidemia. Adult male Wistar rats were pretreated with guarana (12.5, 25 and 50mg/kg/day) and caffeine (0.2mg/kg/day) by gavage for a period of 30days. Simvastatin (0.04mg/kg) was administered as a comparative standard. Acute hyperlipidemia was induced with intraperitoneal injections of 500mg/kg of Poloxamer-407. Memory tests and evaluations of anxiety were performed. The cortex, cerebellum, hippocampus, hypothalamus and striatum were separated to assess acetylcholinesterase activity. Our results revealed that guarana powder was able to reduce the levels of TC and LDL-C in a manner similar to simvastatin. Guarana powder also partially reduced the liver damage caused by hyperlipidemia. Guarana was able to prevent changes in the activity of AChE and improve memory impairment due to hyperlipidemia. Guarana powder may therefore be a source of promising phytochemicals that can be used as adjuvant therapy in the management of hyperlipidemia and cognitive disorders. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Polypyridylruthenium(II complexes exert anti-schistosome activity and inhibit parasite acetylcholinesterases.

    Directory of Open Access Journals (Sweden)

    Madhu K Sundaraneedi

    2017-12-01

    Full Text Available Schistosomiasis affects over 200 million people and there are concerns whether the current chemotherapeutic control strategy (periodic mass drug administration with praziquantel (PZQ-the only licenced anti-schistosome compound is sustainable, necessitating the development of new drugs.We investigated the anti-schistosome efficacy of polypyridylruthenium(II complexes and showed they were active against all intra-mammalian stages of S. mansoni. Two compounds, Rubb12-tri and Rubb7-tnl, which were among the most potent in their ability to kill schistosomula and adult worms and inhibit egg hatching in vitro, were assessed for their efficacy in a mouse model of schistosomiasis using 5 consecutive daily i.v. doses of 2 mg/kg (Rubb12-tri and 10 mg/kg (Rubb7-tnl. Mice treated with Rubb12-tri showed an average 42% reduction (P = 0.009, over two independent trials, in adult worm burden. Liver egg burdens were not significantly decreased in either drug-treated group but ova from both of these groups showed significant decreases in hatching ability (Rubb12-tri-68%, Rubb7-tnl-56% and were significantly morphologically altered (Rubb12-tri-62% abnormal, Rubb7-tnl-35% abnormal. We hypothesize that the drugs exerted their activity, at least partially, through inhibition of both neuronal and tegumental acetylcholinesterases (AChEs, as worms treated in vitro showed significant decreases in activity of these enzymes. Further, treated parasites exhibited a significantly decreased ability to uptake glucose, significantly depleted glycogen stores and withered tubercules (a site of glycogen storage, implying drug-mediated interference in this nutrient acquisition pathway.Our data provide compelling evidence that ruthenium complexes are effective against all intra-mammalian stages of schistosomes, including schistosomula (refractory to PZQ and eggs (agents of disease transmissibility. Further, the results of this study suggest that schistosome AChE is a target of

  13. Heterologous amyloid seeding: revisiting the role of acetylcholinesterase in Alzheimer's disease.

    Directory of Open Access Journals (Sweden)

    Létitia Jean

    2007-07-01

    Full Text Available Neurodegenerative diseases associated with abnormal protein folding and ordered aggregation require an initial trigger which may be infectious, inherited, post-inflammatory or idiopathic. Proteolytic cleavage to generate vulnerable precursors, such as amyloid-beta peptide (Abeta production via beta and gamma secretases in Alzheimer's Disease (AD, is one such trigger, but the proteolytic removal of these fragments is also aetiologically important. The levels of Abeta in the central nervous system are regulated by several catabolic proteases, including insulysin (IDE and neprilysin (NEP. The known association of human acetylcholinesterase (hAChE with pathological aggregates in AD together with its ability to increase Abeta fibrilization prompted us to search for proteolytic triggers that could enhance this process. The hAChE C-terminal domain (T40, AChE(575-614 is an exposed amphiphilic alpha-helix involved in enzyme oligomerisation, but it also contains a conformational switch region (CSR with high propensity for conversion to non-native (hidden beta-strand, a property associated with amyloidogenicity. A synthetic peptide (AChE(586-599 encompassing the CSR region shares homology with Abeta and forms beta-sheet amyloid fibrils. We investigated the influence of IDE and NEP proteolysis on the formation and degradation of relevant hAChE beta-sheet species. By combining reverse-phase HPLC and mass spectrometry, we established that the enzyme digestion profiles on T40 versus AChE(586-599, or versus Abeta, differed. Moreover, IDE digestion of T40 triggered the conformational switch from alpha- to beta-structures, resulting in surfactant CSR species that self-assembled into amyloid fibril precursors (oligomers. Crucially, these CSR species significantly increased Abeta fibril formation both by seeding the energetically unfavorable formation of amyloid nuclei and by enhancing the rate of amyloid elongation. Hence, these results may offer an explanation

  14. Risk of combined exposure of birds to cyanobacterial biomass containing microcystins, acetylcholinesterase inhibitor and anticoagulant.

    Science.gov (United States)

    Ondracek, Karel; Bandouchova, Hana; Damkova, Veronika; Hilscherova, Klara; Kral, Jiri; Osickova, Jitka; Mlcakova, Veronika; Pohanka, Miroslav; Skochova, Hana; Vitula, Frantisek; Treml, Frantisek; Pikula, Jiri

    2012-01-01

    The objective of this study was to examine the hypothesis that a combination of cyanobacterial biomass containing microcystins, acetylcholinesterase inhibitor and anticoagulant can enhance avian toxic effects produced by single exposures only. A total of 48 two-month-old Japanese quails (Coturnix coturnix japonica) with average body weight of 160 g were randomly divided into 8 experimental groups of six birds and sex ratio of 1:1. Experimental groups of control Japanese quails (C) and birds exposed to single and combined sub-lethal doses of paraoxon (P), bromadiolone (B), and microcystins in cyanobacterial biomass (M) included: C, P, P+B, B, B+M, P+M, M, and P+B+M. During the 10-day exposure birds in the respective groups received biomass containing 61.62 µg microcystins daily (i.e. 26.54 µg MC-RR, 7.62 µg MC-YR and 27.39 µg MC-LR), two 250 μg/kg doses of paraoxon, and two 500 mg/kg doses of bromadiolone. Group responses were compared using standard plasma biochemistry and antioxidant/oxidative stress parameters in tissues. While single and double combinations of toxicants induced responses in individual biochemical parameters measured and evaluated using univariate statistical analysis, those in the triple exposure were most extensive. The principal component analysis of antioxidant/oxidative stress parameters (glutathione reductase, lipid peroxidation, and ferric reducing antioxidant power) in tissues (liver, kidney, heart, brain, lungs, gonads, and pectoralis major muscle) clearly separated the triple group (P+B+M) from all single and double exposure groups and the control and indicated thus marked joint effects in the overall pattern of antioxidant/oxidative stress responses of this group. The separation was driven by the modification of the ferric reducing antioxidant power levels in heart and brain and the cardiac lipid peroxidation level, in particular. This experiment contributes to the understanding of the pathogenic mechanisms of combined sub

  15. Rapid eye movement sleep deprivation induces an increase in acetylcholinesterase activity in discrete rat brain regions

    Directory of Open Access Journals (Sweden)

    Benedito M.A.C.

    2001-01-01

    Full Text Available Some upper brainstem cholinergic neurons (pedunculopontine and laterodorsal tegmental nuclei are involved in the generation of rapid eye movement (REM sleep and project rostrally to the thalamus and caudally to the medulla oblongata. A previous report showed that 96 h of REM sleep deprivation in rats induced an increase in the activity of brainstem acetylcholinesterase (Achase, the enzyme which inactivates acetylcholine (Ach in the synaptic cleft. There was no change in the enzyme's activity in the whole brain and cerebrum. The components of the cholinergic synaptic endings (for example, Achase are not uniformly distributed throughout the discrete regions of the brain. In order to detect possible regional changes we measured Achase activity in several discrete rat brain regions (medulla oblongata, pons, thalamus, striatum, hippocampus and cerebral cortex after 96 h of REM sleep deprivation. Naive adult male Wistar rats were deprived of REM sleep using the flower-pot technique, while control rats were left in their home cages. Total, membrane-bound and soluble Achase activities (nmol of thiocholine formed min-1 mg protein-1 were assayed photometrically. The results (mean ± SD obtained showed a statistically significant (Student t-test increase in total Achase activity in the pons (control: 147.8 ± 12.8, REM sleep-deprived: 169.3 ± 17.4, N = 6 for both groups, P<0.025 and thalamus (control: 167.4 ± 29.0, REM sleep-deprived: 191.9 ± 15.4, N = 6 for both groups, P<0.05. Increases in membrane-bound Achase activity in the pons (control: 171.0 ± 14.7, REM sleep-deprived: 189.5 ± 19.5, N = 6 for both groups, P<0.05 and soluble enzyme activity in the medulla oblongata (control: 147.6 ± 16.3, REM sleep-deprived: 163.8 ± 8.3, N = 6 for both groups, P<0.05 were also observed. There were no statistically significant differences in the enzyme's activity in the other brain regions assayed. The present findings show that the increase in Achase activity

  16. Identification of two acetylcholinesterases in Pardosa pseudoannulata and the sensitivity to insecticides.

    Science.gov (United States)

    Zhang, Yixi; Shao, Ying; Jiang, Feng; Li, Jian; Liu, Zewen

    2014-03-01

    Pardosa pseudoannulata is an important predatory enemy against insect pests, such as rice planthoppers and leafhoppers. In order to understand the insecticide selectivity between P. pseudoannulata and insect pests, two acetylcholinesterase genes, Pp-ace1 and Pp-ace2, were cloned from this natural enemy. The putative proteins encoded by Pp-ace1 and Pp-ace2 showed high similarities to insect AChE1 (63% to Liposcelis entomophila AChE1) and AChE2 (36% to Culex quinquefasciatus AChE2) with specific functional motifs, which indicated that two genes might encode AChE1 and AChE2 proteins respectively. The recombinant proteins by expressing Pp-ace1 and Pp-ace2 genes in insect sf9 cells showed high AChE activities. The kinetic parameters, Vmax and Km, of two recombinant AChE proteins were significantly different. The sensitivities to six insecticides were determined in two recombinant AChEs. Pp-AChE1 was more sensitive to all tested insecticides than Pp-AChE2, such as fenobucarb (54 times in Ki ratios), isoprocarb (31 times), carbaryl (13 times) and omethoate (6 times). These results indicated that Pp-AChE1 might be the major synaptic enzyme in the spider. By sequence comparison of P. pseudoannulata and insect AChEs, the key amino acid differences at or close to the functional sites were found. The locations of some key amino acid differences were consistent with the point mutation sites in insect AChEs that were associated with insecticide resistance, such as Phe331 in Pp-AChE2 corresponding to Ser331Phe mutation in Myzus persicae and Aphis gossypii AChE2, which might play important roles in insecticide selectivity between P. pseudoannulata and insect pests. Of course, the direct evidences are needed through further studies. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Identification and Biochemical Properties of Two New Acetylcholinesterases in the Pond Wolf Spider (Pardosa pseudoannulata.

    Directory of Open Access Journals (Sweden)

    Xiangkun Meng

    Full Text Available Acetylcholinesterase (AChE, an important neurotransmitter hydrolase in both invertebrates and vertebrates, is targeted by organophosphorus and carbamate insecticides. In this study, two new AChEs were identified in the pond wolf spider Pardosa pseudoannulata, an important predatory natural enemy of several insect pests. In total, four AChEs were found in P. pseudoannulata (including two AChEs previously identified in our laboratory. The new putative AChEs PpAChE3 and PpAChE4 contain most of the common features of the AChE family, including cysteine residues, choline binding sites, the conserved sequence 'FGESAG' and conserved aromatic residues but with a catalytic triad of 'SDH' rather than 'SEH'. Recombinant enzymes expressed in Sf9 cells showed significant differences in biochemical properties compared to other AChEs, such as the optimal pH, substrate specificity, and catalytic efficiency. Among three test substrates, PpAChE1, PpAChE3 and PpAChE4 showed the highest catalytic efficiency (Vmax/KM for ATC (acetylthiocholine iodide, with PpAChE3 exhibiting a clear preference for ATC based on the VmaxATC/VmaxBTC ratio. In addition, the four PpAChEs were more sensitive to the AChE-specific inhibitor BW284C51, which acts against ATC hydrolysis, than to the BChE-specific inhibitor ISO-OMPA, which acts against BTC hydrolysis, with at least a 8.5-fold difference in IC50 values for each PpAChE. PpAChE3, PpAChE4, and PpAChE1 were more sensitive than PpAChE2 to the tested Carb insecticides, and PpAChE3 was more sensitive than the other three AChEs to the tested OP insecticides. Based on all the results, two new functional AChEs were identified from P. pseudoannulata. The differences in AChE sequence between this spider and insects enrich our knowledge of invertebrate AChE diversity, and our findings will be helpful for understanding the selectivity of insecticides between insects and natural enemy spiders.

  18. Measurement of acetylcholinesterase (AChE) activity in living brain by positron emission tomography (PET)

    Energy Technology Data Exchange (ETDEWEB)

    Irie, Toshiaki [National Inst. of Radiological Sciences, Chiba (Japan)

    1999-07-01

    Central cholinergic neuronal system has been known to be related to learning and memory, and its deficit is found in the brain of Alzheimer's disease (AD) and other degenerative disorders. Postmortem studies have shown that acetylcholinesterase (AChE), one of biochemical markers of central cholinergic nerve system, is consistently reduced in the cerebral cortex of patients with Alzheimer's disease (AD). Non-invasive mapping and/or measuring AChE activity in the living brain by positron emission tomography (PET) would be a useful tool for assessment of cholinergic dysfunction in AD and other disorders, and provide a direct method for validation of therapeutic efficacy of drugs, AChE inhibitors. We have challenged to measure AChE activity using tracers of substrate type, radiolabelled acetylcholine analogs, which are lipophilic enough to go across blood brain barrier and are metabolically trapped by AChE in the brain. The analogs designed, N-methylpiperidyl esters, were evaluated in terms of their metabolic rate and specificity against AChE. Studies examining the response to AChE activity showed metabolic accumulation of some analogs responded well to changes in cortical AChE activity in an animal model of AD. The study was further applied to living human by PET using [{sup 11}C]N-methylpiperidyl-4-acetate (MP4A), which was chosen on the basis of its reactivity and specificity suitable for the human cortical AChE. Regional cerebral metabolic rate of MP4A reflecting AChE activity was quantitatively determined using three compartment model analysis of dynamic PET data and the arterial input function obtained by TLC-radioluminography or plasma samples. The kinetic analyses showed that AChE activities estimated were well agree with those of postmortem examination in cerebral cortices and thalamus in healthy subjects, and that there was significant reduction of cortical AChE activity in patients with AD. The results suggest feasibility of the present method for

  19. The interactions of azure B, a metabolite of methylene blue, with acetylcholinesterase and butyrylcholinesterase

    International Nuclear Information System (INIS)

    Petzer, Anél; Harvey, Brian H.; Petzer, Jacobus P.

    2014-01-01

    Methylene blue (MB) is reported to possess diverse pharmacological actions and is attracting increasing attention for the treatment of neurodegenerative disorders such as Alzheimer's disease. Among the pharmacological actions of MB, is the significant inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). These activities may, at least in part, underlie MB's beneficial effects in Alzheimer's disease. MB is metabolized to yield N-demethylated products of which azure B, the monodemethyl metabolite, is the predominant species. Azure B has been shown to be pharmacologically active and also possesses a variety of biological actions. Azure B therefore may contribute to the pharmacological profile of MB. Based on these considerations, the present study investigates the possibility that azure B may, similar to MB, act as an inhibitor of human AChE and BuChE. The results document that azure B inhibits AChE and BuChE with IC 50 values of 0.486 μM and 1.99 μM, respectively. The results further show that azure B inhibits AChE and BuChE reversibly, and that the modes of inhibition are most likely competitive. Although the AChE and BuChE inhibitory activities of azure B are twofold and fivefold, respectively, less potent than those recorded for MB [IC 50 (AChE) = 0.214 μM; IC 50 (BuChE) = 0.389 μM] under identical conditions, azure B may be a contributor to MB's in vivo activation of the cholinergic system and beneficial effects in Alzheimer's disease. - Highlights: • Methylene blue (MB) is a known inhibitor of AChE and BuChE. • Azure B, the major metabolite of MB, also is an inhibitor of AChE and BuChE. • Azure B may be a contributor to MB's in vivo activation of the cholinergic system. • Azure B may contribute to MB's potential in Alzheimer's disease therapy

  20. Hupresin Retains Binding Capacity for Butyrylcholinesterase and Acetylcholinesterase after Sanitation with Sodium Hydroxide

    Directory of Open Access Journals (Sweden)

    Seda Onder

    2017-10-01

    Full Text Available Hupresin is a new affinity resin that binds butyrylcholinesterase (BChE in human plasma and acetylcholinesterase (AChE solubilized from red blood cells (RBC. Hupresin is available from the CHEMFORASE company. BChE in human plasma binds to Hupresin and is released with 0.1 M trimethylammonium bromide (TMA with full activity and 10–15% purity. BChE immunopurified from plasma by binding to immobilized monoclonal beads has fewer contaminating proteins than the one-step Hupresin-purified BChE. However, when affinity chromatography on Hupresin follows ion exchange chromatography at pH 4.5, BChE is 99% pure. The membrane bound AChE, solubilized from human RBC with 0.6% Triton X-100, binds to Hupresin and remains bound during washing with sodium chloride. Human AChE is not released in significant quantities with non-denaturing solvents, but is recovered in 1% trifluoroacetic acid. The denatured, partially purified AChE is useful for detecting exposure to nerve agents by mass spectrometry. Our goal was to determine whether Hupresin retains binding capacity for BChE and AChE after Hupresin is washed with 0.1 M NaOH. A 2 mL column of Hupresin equilibrated in 20 mM TrisCl pH 7.5 was used in seven consecutive trials to measure binding and recovery of BChE from 100 mL human plasma. Between each trial the Hupresin was washed with 10 column volumes of 0.1 M sodium hydroxide. A similar trial was conducted with red blood cell AChE in 0.6% Triton X-100. It was found that the binding capacity for BChE and AChE was unaffected by washing Hupresin with 0.1 M sodium hydroxide. Hupresin could be washed with sodium hydroxide at least seven times without losing binding capacity.

  1. Unmasking tandem site interaction in human acetylcholinesterase. Substrate activation with a cationic acetanilide substrate.

    Science.gov (United States)

    Johnson, Joseph L; Cusack, Bernadette; Davies, Matthew P; Fauq, Abdul; Rosenberry, Terrone L

    2003-05-13

    Acetylcholinesterase (AChE) contains a narrow and deep active site gorge with two sites of ligand binding, an acylation site (or A-site) at the base of the gorge, and a peripheral site (or P-site) near the gorge entrance. The P-site contributes to catalytic efficiency by transiently binding substrates on their way to the acylation site, where a short-lived acyl enzyme intermediate is produced. A conformational interaction between the A- and P-sites has recently been found to modulate ligand affinities. We now demonstrate that this interaction is of functional importance by showing that the acetylation rate constant of a substrate bound to the A-site is increased by a factor a when a second molecule of substrate binds to the P-site. This demonstration became feasible through the introduction of a new acetanilide substrate analogue of acetylcholine, 3-(acetamido)-N,N,N-trimethylanilinium (ATMA), for which a = 4. This substrate has a low acetylation rate constant and equilibrates with the catalytic site, allowing a tractable algebraic solution to the rate equation for substrate hydrolysis. ATMA affinities for the A- and P-sites deduced from the kinetic analysis were confirmed by fluorescence titration with thioflavin T as a reporter ligand. Values of a >1 give rise to a hydrolysis profile called substrate activation, and the AChE site-specific mutant W86F, and to a lesser extent wild-type human AChE itself, showed substrate activation with acetylthiocholine as the substrate. Substrate activation was incorporated into a previous catalytic scheme for AChE in which a bound P-site ligand can also block product dissociation from the A-site, and two additional features of the AChE catalytic pathway were revealed. First, the ability of a bound P-site ligand to increase the substrate acetylation rate constant varied with the structure of the ligand: thioflavin T accelerated ATMA acetylation by a factor a(2) of 1.3, while propidium failed to accelerate. Second, catalytic rate

  2. How does huperzine A enter and leave the binding gorge of acetylcholinesterase? Steered molecular dynamics simulations.

    Science.gov (United States)

    Xu, Yechun; Shen, Jianhua; Luo, Xiaomin; Silman, Israel; Sussman, Joel L; Chen, Kaixian; Jiang, Hualiang

    2003-09-17

    The entering and leaving processes of Huperzine A (HupA) binding with the long active-site gorge of Torpedo californica acetylcholinesterase (TcAChE) have been investigated by using steered molecular dynamics simulations. The analysis of the force required along the pathway shows that it is easier for HupA to bind to the active site of AChE than to disassociate from it, which for the first time interprets at the atomic level the previous experimental result that unbinding process of HupA is much slower than its binding process to AChE. The direct hydrogen bonds, water bridges, and hydrophobic interactions were analyzed during two steered molecular dynamics (SMD) simulations. Break of the direct hydrogen bond needs a great pulling force. The steric hindrance of bottleneck might be the most important factor to produce the maximal rupture force for HupA to leave the binding site but it has a little effect on the binding process of HupA with AChE. Residue Asp72 forms a lot of water bridges with HupA leaving and entering the AChE binding gorge, acting as a clamp to take out HupA from or put HupA into the active site. The flip of the peptide bond between Gly117 and Gly118 has been detected during both the conventional MD and SMD simulations. The simulation results indicate that this flip phenomenon could be an intrinsic property of AChE and the Gly117-Gly118 peptide bond in both HupA bound and unbound AChE structures tends to adopt the native enzyme structure. At last, in a vacuum the rupture force is increased up to 1500 pN while in water solution the greatest rupture force is about 800 pN, which means water molecules in the binding gorge act as lubricant to facilitate HupA entering or leaving the binding gorge.

  3. The application of HPLC with on-line coupled UV/MS-biochemical detection for isolation of an acetylcholinesterase inhibitor from Narcissus 'Sir Winston Churchill'

    NARCIS (Netherlands)

    Ingkaninan, K.; Hazekamp, A.; de Best, C.M.; Irth, H.; Tjaden, U.R.; van der Heijden, R.; van der Greef, J.; Verpoorte, R.

    2000-01-01

    An HPLC with on-line coupled UV/MS-biochemical detection method for acetylcholinesterase (AChE) inhibitors in natural sources has been developed. The potential of this method is shown by the isolation of a new AChE inhibitor from the alcoholic extract of Narcissus 'Sir Winston Churchill'. Combining

  4. Synthesis of a Pilot Scale Library of 4-amino-2 (diethylaminomethyl) phenol (ADOC) Analogues for Testing of Organophosphate-Inhibited Acetylcholinesterase Reactivation Ability

    Science.gov (United States)

    2018-02-01

    Inhibited Acetylcholinesterase Reactivation Ability Zachary Canter Kevin Martin Michael Hepperle February 2018 Approved for public release; distribution...PROGRAM ELEMENT NUMBER 6. AUTHOR(S) Canter, Z., Martin , K., Hepperle, M. 5d. PROJECT NUMBER 5e. TASK NUMBER 5f. WORK UNIT NUMBER 7. PERFORMING

  5. Chlamydia psittaci and C. avium in feral pigeon (Columba livia domestica) droppings in two cities in the Netherlands.

    Science.gov (United States)

    Burt, Sara A; Röring, Romy E; Heijne, Marloes

    2018-06-05

    Feral pigeons (Columba livia domestica) live and breed in many city centres and contact with their droppings can be a hazard for human health if the birds carry Chlamydia psittaci. The aim of this study was to establish whether pigeon droppings in two Dutch cities (Utrecht and Haarlem) contain C. psittaci and/or C. avium, which could be a potential hazard for transmission to humans. In May 2017 seven feral pigeon 'hot spots' with between 5 and 40+ pigeons present were identified in two cities by visual observations over two days. During the following ten days fresh droppings were collected at these hot spots and the samples were pooled per three droppings to achieve 40-41 samples per city. Samples were analysed for Chlamydia DNA with a broad range 23S Chlamydiaceae Real-Time PCR and positive samples were tested with a specific C. psittaci and C. avium Real-Time PCR. Positive C. psittaci samples were genotyped. C. psittaci and C. avium were detected in both cities. For C. psittaci the prevalences in Utrecht and Haarlem were 2.4% and 7.5%, respectively; for C. avium 36.6% and 20.0%, respectively. One sample contained both species. All C. psittaci samples belonged to genotype B. C. psittaci and C. avium are present in feral pigeon droppings in Utrecht and Haarlem. Human contact with droppings from infected pigeons or inhalation of dust from dried droppings represent a potential hazard to public health.

  6. Prunus domestica pathogenesis-related protein-5 activates the defense response pathway and enhances the resistance to fungal infection.

    Directory of Open Access Journals (Sweden)

    Ashraf El-kereamy

    Full Text Available Pathogenesis-related protein-5 (PR-5 has been implicated in plant disease resistance and its antifungal activity has been demonstrated in some fruit species. However, their roles, especially their interactions with the other defense responses in plant cells, are still not fully understood. In this study, we have cloned and characterized a new PR-5 cDNA named PdPR5-1 from the European plum (Prunus domestica. Expression of PdPR5-1 was studied in different cultivars varying in resistance to the brown rot disease caused by the necrotrophic fungus Monilinia fructicola. In addition transgenic Arabidopsis, ectopically expressing PdPR5-1 was used to study its role in other plant defense responses after fungal infection. We show that the resistant cultivars exhibited much higher levels of transcripts than the susceptible cultivars during fruit ripening. However, significant rise in the transcript levels after infection with M. fructicola was observed in the susceptible cultivars too. Transgenic Arabidopsis plants exhibited more resistance to Alternaria brassicicola. Further, there was a significant increase in the transcripts of genes involved in the phenylpropanoid biosynthesis pathway such as phenylalanine ammonia-lyase (PAL and phytoalexin (camalexin pathway leading to an increase in camalexin content after fungal infection. Our results show that PdPR5-1 gene, in addition to its anti-fungal properties, has a possible role in activating other defense pathways, including phytoalexin production.

  7. Foliar application of amino acids modulates aroma components of 'FUJI' apple (malus domestica L.)

    International Nuclear Information System (INIS)

    Gou, W.; Zhang, L.; Chen, F.; Cui, Z.; Zhao, Y.; Zheng, P.; Tian, L.; Zhang, L.; Zhang, C.

    2015-01-01

    Volatile flavor compounds play a key role in determining the perception and acceptability as well as enhancing market competitiveness of apple (Malus domestica L.). In our study, we evaluated the effects of foliar-applied four different amino acids, i.e. leucine (Leu), isoleucine (Ile), valine (Val) and alanine (Ala), on aroma components and two key enzymes activities involved in aroma metabolism of Fuji apple. The total amount of aromatic components under Ala treatment was significantly higher than those under other treatments. There was a considerable increase in total aroma content, including hexanal, 2-methyl-butanol, nonanal, (E)-2-hexenal, methyleugenol, ethyl acetate, butanoic acid-pentyl ester, butanoic acid-hexyl ester, butyric acid ethyl ester, acetic acid-2-methyl-butyl ester, treated with spraying amino acids compared with the control. More specifically, hexanal, 2-methyl-butanol, methyleugenol and acetic acid-2-methyl-butyl ester exhibited a greater substantial increase of their contents than those of in other ingredients. However, butanoic acid-2-methyl-2-methyl butyl ester maintained a highest level among all aroma components regardless of different amino acids application. Furthermore, the activities of alcohol dehydrogenase (ADH) and alcohol acyltransferase (AAT) were much higher under Ala treatment than those under other treatments. We concluded that foliar-applied organic nitrogen (N), especially for Ala, can improve aroma metabolism and it could be used in production to enhance fruit quality on a commercial scale. (author)

  8. Involvement of plasma membrane peroxidases and oxylipin pathway in the recovery from phytoplasma disease in apple (Malus domestica).

    Science.gov (United States)

    Patui, Sonia; Bertolini, Alberto; Clincon, Luisa; Ermacora, Paolo; Braidot, Enrico; Vianello, Angelo; Zancani, Marco

    2013-06-01

    Apple trees (Malus domestica Borkh.) may be affected by apple proliferation (AP), caused by 'Candidatus Phytoplasma mali'. Some plants can spontaneously recover from the disease, which implies the disappearance of symptoms through a phenomenon known as recovery. In this article it is shown that NAD(P)H peroxidases of leaf plasma membrane-enriched fractions exhibited a higher activity in samples from both AP-diseased and recovered plants. In addition, an increase in endogenous SA was characteristic of the symptomatic plants, since its content increased in samples obtained from diseased apple trees. In agreement, phenylalanine ammonia lyase (PAL) activity, a key enzyme of the phenylpropanoid pathway, was increased too. Jasmonic acid (JA) increased only during recovery, in a phase subsequent to the pathological state, and in concomitance to a decline of salicylic acid (SA). Oxylipin pathway, responsible for JA synthesis, was not induced during the development of AP-disease, but it appeared to be stimulated when the recovery occurred. Accordingly, lipoxygenase (LOX) activity, detected in plasma membrane-enriched fractions, showed an increase in apple leaves obtained from recovered plants. This enhancement was paralleled by an increase of hydroperoxide lyase (HPL) activity, detected in leaf microsomes, albeit the latter enzyme was activated in either the disease or recovery conditions. Hence, a reciprocal antagonism between SA- and JA-pathways could be suggested as an effective mechanism by which apple plants react to phytoplasma invasions, thereby providing a suitable defense response leading to the establishment of the recovery phenomenon. Copyright © Physiologia Plantarum 2012.

  9. Down-regulation of POLYGALACTURONASE1 alters firmness, tensile strength and water loss in apple (Malus x domestica) fruit.

    Science.gov (United States)

    Atkinson, Ross G; Sutherland, Paul W; Johnston, Sarah L; Gunaseelan, Kularajathevan; Hallett, Ian C; Mitra, Deepali; Brummell, David A; Schröder, Roswitha; Johnston, Jason W; Schaffer, Robert J

    2012-08-02

    While there is now a significant body of research correlating apple (Malus x domestica) fruit softening with the cell wall hydrolase ENDO-POLYGALACTURONASE1 (PG1), there is currently little knowledge of its physiological effects in planta. This study examined the effect of down regulation of PG1 expression in 'Royal Gala' apples, a cultivar that typically has high levels of PG1, and softens during fruit ripening. PG1-suppressed 'Royal Gala' apples harvested from multiple seasons were firmer than controls after ripening, and intercellular adhesion was higher. Cell wall analyses indicated changes in yield and composition of pectin, and a higher molecular weight distribution of CDTA-soluble pectin. Structural analyses revealed more ruptured cells and free juice in pulled apart sections, suggesting improved integrity of intercellular connections and consequent cell rupture due to failure of the primary cell walls under stress. PG1-suppressed lines also had reduced expansion of cells in the hypodermis of ripe apples, resulting in more densely packed cells in this layer. This change in morphology appears to be linked with reduced transpirational water loss in the fruit. These findings confirm PG1's role in apple fruit softening and suggests that this is achieved in part by reducing cellular adhesion. This is consistent with previous studies carried out in strawberry but not with those performed in tomato. In apple PG1 also appears to influence other fruit texture characters such as juiciness and water loss.

  10. Evaluation of the hormonal state of columnar apple trees (Malus x domestica) based on high throughput gene expression studies.

    Science.gov (United States)

    Krost, Clemens; Petersen, Romina; Lokan, Stefanie; Brauksiepe, Bastienne; Braun, Peter; Schmidt, Erwin R

    2013-02-01

    The columnar phenotype of apple trees (Malus x domestica) is characterized by a compact growth habit with fruit spurs instead of lateral branches. These properties provide significant economic advantages by enabling high density plantings. The columnar growth results from the presence of a dominant allele of the gene Columnar (Co) located on chromosome 10 which can appear in a heterozygous (Co/co) or homozygous (Co/Co) state. Although two deep sequencing approaches could shed some light on the transcriptome of columnar shoot apical meristems (SAMs), the molecular mechanisms of columnar growth are not yet elaborated. Since the influence of phytohormones is believed to have a pivotal role in the establishment of the phenotype, we performed RNA-Seq experiments to study genes associated with hormone homeostasis and clearly affected by the presence of Co. Our results provide a molecular explanation for earlier findings on the hormonal state of columnar apple trees. Additionally, they allow hypotheses on how the columnar phenotype might develop. Furthermore, we show a statistically approved enrichment of differentially regulated genes on chromosome 10 in the course of validating RNA-Seq results using additional gene expression studies.

  11. Estrogenic Effect of 70% Ethanol Turmeric (Curcuma domestica Val. Extract on Ovariectomized Female Mice (Mus musculus L.

    Directory of Open Access Journals (Sweden)

    A.N. Dewi

    2007-11-01

    Full Text Available The influence of extract turmeric (Curcuma domestica Val. on endometrium thickness, vaginal epithelium, mammary gland, and protein of estrogen receptor of ovariectomized mice was examined. Twenty five ovariectomized mice which were divided into five groups, were treated by ethynilestradiol (8,4 x 10-3 g, aquades (10 ml, and turmeric extract at doses 230 mg/kg b.w.; 310 mg/kg b.w.; and 390 mg/kg b.w. for eight days. At the end of experiments the mice were killed, then the uterus, vagina, and mammae were removed and the wet weight of uterus was recorded. Uterus, vagina, and mammae were examined histologically. Estrogen receptor protein from uterus were analized by using SDS-PAGE. One way anava test showed that turmeric extract at doses 310 mg/kg b.w. and 390 mg/kg b.w give estrogenic effect on vaginal ephitelium, endometrium thickness, and diametre of mammary glands. SDS-PAGE analysis showed there were differences in protein concentration between control and treatment groups which were seen in the thickness of the bands. Estrogen receptor band could be detected in sampel of treatment groups at molecular weight 45 kDa.

  12. Suppressing Sorbitol Synthesis Substantially Alters the Global Expression Profile of Stress Response Genes in Apple (Malus domestica) Leaves.

    Science.gov (United States)

    Wu, Ting; Wang, Yi; Zheng, Yi; Fei, Zhangjun; Dandekar, Abhaya M; Xu, Kenong; Han, Zhenhai; Cheng, Lailiang

    2015-09-01

    Sorbitol is a major product of photosynthesis in apple (Malus domestica) that is involved in carbohydrate metabolism and stress tolerance. However, little is known about how the global transcript levels in apple leaves respond to decreased sorbitol synthesis. In this study we used RNA sequencing (RNA-seq) profiling to characterize the transcriptome of leaves from transgenic lines of the apple cultivar 'Greensleeves' exhibiting suppressed expression of aldose-6-phosphate reductase (A6PR) to gain insights into sorbitol function and the consequences of decreased sorbitol synthesis on gene expression. We observed that, although the leaves of the low sorbitol transgenic lines accumulate higher levels of various primary metabolites, only very limited changes were found in the levels of transcripts associated with primary metabolism. We suggest that this is indicative of post-transcriptional and/or post-translational regulation of primary metabolite accumulation and central carbon metabolism. However, we identified significantly enriched gene ontology terms belonging to the 'stress related process' category in the antisense lines (P-value sorbitol plays a role in the responses of apple trees to abiotic and biotic stresses. © The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  13. Preliminary studies on the reaction of growing geese (Anser anser f. domestica) to the proximity of wind turbines.

    Science.gov (United States)

    Mikolajczak, J; Borowski, S; Marć-Pieńkowska, J; Odrowaz-Sypniewska, G; Bernacki, Z; Siódmiak, J; Szterk, P

    2013-01-01

    Wind farms produce electricity without causing air pollution and environmental degradation. Unfortunately, wind turbines are a source of infrasound, which may cause a number of physiological effects, such as an increase in cortisol and catecholamine secretion. The impact of infrasound noise, emitted by wind turbines, on the health of geese and other farm animals has not previously been evaluated. Therefore, the aim of this study was to determine the effect of noise, generated by wind turbines, on the stress parameters (cortisol) and the weight gain of geese kept in surrounding areas. The study consisted of 40 individuals of 5-week-old domestic geese Anser anser f domestica, divided into 2 equal groups. The first experimental gaggle (I) remained within 50 m from turbine and the second one (II) within 500 m. During the 12 weeks of the study, noise measurements were also taken. Weight gain and the concentration of cortisol in blood were assessed and significant differences in both cases were found. Geese from gaggle I gained less weight and had a higher concentration of cortisol in blood, compared to individuals from gaggle II. Lower activity and some disturbing changes in behavior of animals from group I were noted. Results of the study suggest a negative effect of the immediate vicinity of a wind turbine on the stress parameters of geese and their productivity.

  14. The effect of apple (Malus Domestica juice on the damage of mice liver cells due to paracetamol treatment

    Directory of Open Access Journals (Sweden)

    Anthony Hartanto

    2009-07-01

    Full Text Available The liver is an important organ for body metabolism process. Liver disease is one of serious health problems in developing countries including Indonesia. Liver damage is caused by viral infection, toxic agent exposure (medications, alcohol, hormonal disturbance, neoplasm and autoimmune diseases. The use of high dose paracetamol to reduce pain also leads to liver damage. Apple (Malus domestica juice is a natural anti oxidant agent. This laboratory experimental study was performed to discover the effect of giving apple juice on damaged cell regeneration due to the use of paracetamol. The study was performed in 21 male mice from Swiss-Webster strain that were divided into group I, II, and III. Group, I served as control while group II received 1 mg/ml paracetamol dose for 5 days and Group III received 1 mg/ml paracetamol for 5 days and 1 ml of apple juice on the 5th to 10th day. The observation of the mice liver cells was conducted using a light microscope with 400x magnification to get the number of necrotic liver cells per view field. The results of this study showed a difference in the number of necrotic liver cells between Group II and III. ANOVA statistical test ( = 0.05 concluded that apple juice significantly helps regeneration process in damaged liver cells caused by paracetamol.

  15. The role of the oximes HI-6 and HS-6 inside human acetylcholinesterase inhibited with nerve agents: a computational study.

    Science.gov (United States)

    Cuya, Teobaldo; Gonçalves, Arlan da Silva; da Silva, Jorge Alberto Valle; Ramalho, Teodorico C; Kuca, Kamil; C C França, Tanos

    2017-10-27

    The oximes 4-carbamoyl-1-[({2-[(E)-(hydroxyimino) methyl] pyridinium-1-yl} methoxy) methyl] pyridinium (known as HI-6) and 3-carbamoyl-1-[({2-[(E)-(hydroxyimino) methyl] pyridinium-1-yl} methoxy) methyl] pyridinium (known as HS-6) are isomers differing from each other only by the position of the carbamoyl group on the pyridine ring. However, this slight difference was verified to be responsible for big differences in the percentual of reactivation of acetylcholinesterase (AChE) inhibited by the nerve agents tabun, sarin, cyclosarin, and VX. In order to try to find out the reason for this, a computational study involving molecular docking, molecular dynamics, and binding energies calculations, was performed on the binding modes of HI-6 and HS-6 on human AChE (HssAChE) inhibited by those nerve agents.

  16. Chitooligosaccharides suppress the level of protein expression and acetylcholinesterase activity induced by Abeta25-35 in PC12 cells.

    Science.gov (United States)

    Lee, Sang-Hoon; Park, Jin-Sook; Kim, Se-Kwon; Ahn, Chang-Bum; Je, Jae-Young

    2009-02-01

    Clinical applications of acetylcholinesterase (AChE) inhibitors are widespread in Alzheimer's sufferers in order to activate central cholinergic system and alleviate cognitive deficits by inhibiting the hydrolysis of acetylcholine. In this study, six kinds of chitooligosaccharides (COSs) with different molecular weight and degree of deacetylation were examined for their inhibitory effects against AChE. The 90-COSs exhibited potent AChE inhibitory activities compared to 50-COSs, while 90-MMWCOS (1000-5000 Da) in the 90-COSs showed the highest activity. Cell culture experiment revealed that 90-MMWCOS suppressed the level of AChE protein expression and AChE activity induced by Abeta(25-35) in PC12 cell lines.

  17. Synthesis and biological evaluation of phloroglucinol derivatives possessing α-glycosidase, acetylcholinesterase, butyrylcholinesterase, carbonic anhydrase inhibitory activity.

    Science.gov (United States)

    Burmaoglu, Serdar; Yilmaz, Ali O; Taslimi, Parham; Algul, Oztekin; Kilic, Deryanur; Gulcin, Ilhami

    2018-02-01

    A series of novel phloroglucinol derivatives were designed, synthesized, characterized spectroscopically and tested for their inhibitory activity against selected metabolic enzymes, including α-glycosidase, acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and human carbonic anhydrase I and II (hCA I and II). These compounds displayed nanomolar inhibition levels and showed K i values of 1.14-3.92 nM against AChE, 0.24-1.64 nM against BChE, 6.73-51.10 nM against α-glycosidase, 1.80-5.10 nM against hCA I, and 1.14-5.45 nM against hCA II. © 2018 Deutsche Pharmazeutische Gesellschaft.

  18. Effect of thermal stress and water deprivation on the acetylcholinesterase activity of the pig brain and hypophyses

    Science.gov (United States)

    Adejumo, D. O.; Egbunike, G. N.

    1988-06-01

    The effects of direct exposure of boars to thermal stress for 1 h daily for 5 days and to acute water deprivation for 24 or 48 h were studied on the acetylcholinesterase (AChE) activity of porcine brain and hypophysial regions. Mean ambient temperatures, respiratory rates and rectal temperatures in the open were significantly higher than inside the pen. Heat stress induced a rise in AChE activities in the pons, cerebellum, amygdala, hippocampus, hypothalamus, mid-brain and medulla oblongata. However, no significant changes were observed in the cerebral cortex, adenohypophysis and neurohypophysis. Water deprivation significantly ( Pmedulla oblongata were comparable to the amygdala in this respect. The adenohypophysis and neurohypophysis were relatively unaffected.

  19. Novel hybrids of oxoisoaporphine-tryptamine as acetylcholinesterase-induced β-amyloid aggregation inhibitors with improved antioxidant properties

    Directory of Open Access Journals (Sweden)

    Zhao Hai-Tao

    2015-01-01

    Full Text Available A series of dual binding site acetylcholinesterase (AChE inhibitors have been designed, synthesized, and tested for their antioxidant ability and inhibitory potency on AChE and AChE-induced b-amyloid (Ab aggregation. The new hybrids consist of a unit of 1-azabenzanthrone and a tryptamine or its derivative, connected through a a,w - alkyldiamide bridge. These hybrids exhibit moderate AChE inhibitory activity with IC50 values in the micromolar range and significant in vitro inhibitory activity toward the AChE-induced Ab aggregation. Moreover, six out of the nine hybrids of this series exhibit a higher oxygen radical absorbance capacity than trolox, which makes them promising anti-Alzheimer drug candidates.

  20. Is subnanomolar binding affinity required for the in vivo imaging of acetylcholinesterase? Studies on 18F-labeled G379

    International Nuclear Information System (INIS)

    Lee, Sang-Yoon; Choe, Yearn Seong; Ryu, Eun Kyoung; Iimura, Yoichi; Choi, Yong; Lee, Kyung-Han; Kim, Byung-Tae

    2006-01-01

    Acetylcholinesterase (AChE) is an important cholinergic marker of Alzheimer's disease (AD) and shows reduced activity in postmortem AD brain tissues. 1-(4-Fluorobenzyl)-4-[(5,6-dimethoxy-1-oxoindan-2-fluoro-2-yl)methyl] piperidine (G379, ), an AChE inhibitor with a subnanomolar IC 5 (0.56 nM), was prepared as a 18 F-labeled radioligand ([ 18 F]) and evaluated in mice. Metabolism studies of [ 18 F] showed no metabolites in the mouse brain. Tissue distribution studies demonstrated its uniform regional distribution in the mouse brain, suggesting that this radioligand is not suitable for the in vivo imaging of AChE. This result along with reports on radiolabeled N-benzylpiperidine lactam benzisoxazole (IC 5 5 >1 nM) suggested that a subnanomolar IC 5 may not be the only important factor in determining the suitability of a radioligand for in vivo studies of AChE

  1. Production and enzyme engineerinq of human acetylcholinesterase and its mutant derivatives. Midterm report, 15 January 1993-15 July 1994

    Energy Technology Data Exchange (ETDEWEB)

    Shafferman, A.

    1994-07-15

    Specificity determinants of human acetylcholinesterase (HuAChE) towards ligands (substrate and some reversible and irreversible inhibitors) were identified by combination of site-directed mutagenesis, molecular modeling and kinetic studies with enzymes mutated in active center residues Trp86, Glu202, Trp286, Phe295, Phe297, Tyr337, Phe338 and Glu450. Thus, the anionic and hydrophobic subsites as well as the acyl pocket were identified. Enzymes with resistance to OP aging were engineered.The role of N-glycosylation in the function, biosynthesis and stability of HuAChE was examined by site-directed mutagenesis (Asn to GIn substitution) of the three potential N glycosylation sites, Asn265, Asn350 and Asn464. Large scale preparation of recombinant HuAChE was performed utilizing the microcarrier technology. Over 500 milligrams of enzyme was prepared for x-ray crystallography.

  2. 7-Methoxytacrine-p-Anisidine Hybrids as Novel Dual Binding Site Acetylcholinesterase Inhibitors for Alzheimer’s Disease Treatment

    Directory of Open Access Journals (Sweden)

    Jan Korabecny

    2015-12-01

    Full Text Available Alzheimer’s disease (AD is a debilitating progressive neurodegenerative disorder that ultimately leads to the patient’s death. Despite the fact that novel pharmacological approaches endeavoring to block the neurodegenerative process are still emerging, none of them have reached use in clinical practice yet. Thus, palliative treatment represented by acetylcholinesterase inhibitors (AChEIs and memantine are still the only therapeutics used. Following the multi-target directed ligands (MTDLs strategy, herein we describe the synthesis, biological evaluation and docking studies for novel 7-methoxytacrine-p-anisidine hybrids designed to purposely target both cholinesterases and the amyloid cascade. Indeed, the novel derivatives proved to be effective non-specific cholinesterase inhibitors showing non-competitive AChE inhibition patterns. This compounds’ behavior was confirmed in the subsequent molecular modeling studies.

  3. [Distribution of acetylcholinesterase activity in the digestive system of the gastropod molluscs Littorina littorea and Achatina fulica].

    Science.gov (United States)

    Zaĭtseva, O V; Kuznetsova, T V

    2008-01-01

    With the use of the histochemical procedure for the demonstration of acetylcholinesterase (AchE) activity, the distribution cholinergic regulatory elements was studied in the esophagus, the pharynx, the stomach, the liver (the digestive gland) and the intestine in sea and terrestrial gastropod molluscs that differed in their general organization level, lifestyle, habitat and feeding type. In both molluscs, all the parts of the digestive tract contained the significant amount of intraepithelial AchE-positive cells of the open type, single subepithelial neurons and the nervous fibers localized among the muscle cells of the wall of the organs. The basal processes of the AchE-positive intraepithelial cells were shown to form the intraepithelial nerve plexus and to pass under the epithelium. The peculiarities and common principles in the distribution of the nervous elements detected, their possible function and the regulatory role in the digestion in gastropod molluscs and other animals are discussed.

  4. Acetylcholinesterase immobilized capillary reactors coupled to protein coated magnetic beads: A new tool for plant extract ligand screening

    Science.gov (United States)

    Vanzolini, Kenia Lourenço; Jiang, Zhengjin; Zhang, Xiaoqi; Vieira, Lucas Campos Curcino; Corrêa, Arlene Gonçalvez; Cardoso, Carmen Lucia; Cass, Quezia Bezerra; Moaddel, Ruin

    2013-01-01

    The use of immobilized capillary enzyme reactors (ICERs) and enzymes coated to magnetic beads ((NT or CT)-MB) for ligand screening has been adopted as a new technique of high throughput screening (HTS). In this work the selected target was the enzyme acetylcholinesterase (AChE), which acts on the central nervous system and is a validated target for the treatment of Alzheimer’s disease, as well as for new insecticides. A new approach for the screening of plant extracts was developed based on the ligand fishing experiments and zonal chromatography. For that, the magnetic beads were used for the ligand fishing experiments and capillary bioreactors for the activity assays. The latter was employed also under non-linear conditions to determine the affinity constants of known ligands, for the first time, as well as for the active fished ligand. PMID:24148457

  5. Acetylcholinesterase (AChE) gene modification in transgenic animals: functional consequences of selected exon and regulatory region deletion.

    Science.gov (United States)

    Camp, Shelley; Zhang, Limin; Marquez, Michael; de la Torre, Brian; Long, Jeffery M; Bucht, Goran; Taylor, Palmer

    2005-12-15

    AChE is an alternatively spliced gene. Exons 2, 3 and 4 are invariantly spliced, and this sequence is responsible for catalytic function. The 3' alternatively spliced exons, 5 and 6, are responsible for AChE disposition in tissue [J. Massoulie, The origin of the molecular diversity and functional anchoring of cholinesterases. Neurosignals 11 (3) (2002) 130-143; Y. Li, S. Camp, P. Taylor, Tissue-specific expression and alternative mRNA processing of the mammalian acetylcholinesterase gene. J. Biol. Chem. 268 (8) (1993) 5790-5797]. The splice to exon 5 produces the GPI anchored form of AChE found in the hematopoietic system, whereas the splice to exon 6 produces a sequence that binds to the structural subunits PRiMA and ColQ, producing AChE expression in brain and muscle. A third alternative RNA species is present that is not spliced at the 3' end; the intron 3' of exon 4 is used as coding sequence and produces the read-through, unanchored form of AChE. In order to further understand the role of alternative splicing in the expression of the AChE gene, we have used homologous recombination in stem cells to produce gene specific deletions in mice. Alternatively and together exon 5 and exon 6 were deleted. A cassette containing the neomycin gene flanked by loxP sites was used to replace the exon(s) of interest. Tissue analysis of mice with exon 5 deleted and the neomycin cassette retained showed very low levels of AChE expression, far less than would have been anticipated. Only the read-through species of the enzyme was produced; clearly the inclusion of the selection cassette disrupted splicing of exon 4 to exon 6. The selection cassette was then deleted in exon 5, exon 6 and exons 5 + 6 deleted mice by breeding to Ella-cre transgenic mice. AChE expression in serum, brain and muscle has been analyzed. Another AChE gene targeted mouse strain involving a region in the first intron, found to be critical for AChE expression in muscle cells [S. Camp, L. Zhang, M. Marquez, B

  6. Acetylcholinesterase inhibitors for the treatment of Wernicke-Korsakoff syndrome--three further cases show response to donepezil.

    Science.gov (United States)

    Cochrane, Murray; Cochrane, Ashley; Jauhar, Pramod; Ashton, Elizabeth

    2005-01-01

    Three patients diagnosed with Wernicke-Korsakoff syndrome were treated with the acetylcholinesterase inhibitor, donepezil, for periods of 6 to 8 months. Cognitive testing [Alzheimer's disease assessment scale-cognitive subscale (ADAS-Cog), Mini-mental state examination (MMSE), Clock drawing test and six item 2 min recall] and carer questionnaires [Informant Questionnaire (IQ Code), Neuropsychiatric inventory scale (NPI)] were performed at baseline, mid- and endpoint of the treatment period and post-discontinuation. Progressive partial improvement occurred in cognitive measurements through the treatment period, some of which was sustained after discontinuing donepezil. Carer questionnaires also indicated improvement. Confounding factors necessitate caution when attributing improvements to the medication, but these cases suggest that this option merits further investigation.

  7. Geissoschizine methyl ether, a corynanthean-type indole alkaloid from Uncaria rhynchophylla as a potential acetylcholinesterase inhibitor.

    Science.gov (United States)

    Yang, Zhong-Duo; Duan, Dong-Zhu; Du, Juan; Yang, Ming-Jun; Li, Shuo; Yao, Xiao-Jun

    2012-01-01

    Geissoschizine methyl ether (1), a newly discovered strong acetylcholinesterase (AChE) inhibitor, along with six weakly active alkaloids, vallesiachotamine (2), hisuteine (3), hirsutine (4), isorhynchophylline (5), cisocorynoxeine (6) and corynoxeine (7) have been isolated from Uncaria rhynchophylla. Geissoschizine methyl ether (1) inhibited 50% of AChE activity at concentrations of 3.7 ± 0.3 µg mL(-1) while the IC(50) value of physostigmine as a standard was 0.013 ± 0.002 µg mL(-1). The mode of AChE inhibition by 1 was reversible and non-competitive. In addition, molecular modelling was performed to explore the binding mode of inhibitor 1 at the active site of AChE.

  8. Energetics of Ortho-7 (oxime drug translocation through the active-site gorge of tabun conjugated acetylcholinesterase.

    Directory of Open Access Journals (Sweden)

    Vivek Sinha

    Full Text Available Oxime drugs translocate through the 20 Å active-site gorge of acetylcholinesterase in order to liberate the enzyme from organophosphorus compounds' (such as tabun conjugation. Here we report bidirectional steered molecular dynamics simulations of oxime drug (Ortho-7 translocation through the gorge of tabun intoxicated enzyme, in which time dependent external forces accelerate the translocation event. The simulations reveal the participation of drug-enzyme hydrogen bonding, hydrophobic interactions and water bridges between them. Employing nonequilibrium theorems that recovers the free energy from irreversible work done, we reconstruct potential of mean force along the translocation pathway such that the desired quantity represents an unperturbed system. The potential locates the binding sites and barriers for the drug to translocate inside the gorge. Configurational entropic contribution of the protein-drug binding entity and the role of solvent translational mobility in the binding energetics is further assessed.

  9. Bovine nucleus caudatus acetylcholinesterase: active site determination and investigation of a dimeric form obtained by selective proteolysis

    Energy Technology Data Exchange (ETDEWEB)

    Landauer, P.; Ruess, K.P.; Lieflaender, M.

    1984-09-01

    The number of catalytic subunits of purified bovine nucleus caudatus acetylcholinesterase (E.C. 3.1.1.7) has been determined by active site labelling with (3H)diisopropyl fluorophosphate ((3H)DFP). The 10.5 S, 16 S, and 20 S forms were estimated to contain two, four, and six active sites, respectively, per molecule. A 4.8 S form, which showed a weak amphiphile-dependent activity behavior, was obtained by selective proteolytic digestion with pronase. The inability of the purified 4.8 S form to aggregate after detergent removal, and the molecular mass in the range of 130-165 kD under nondenaturating conditions, indicate that this form is a dimeric form, lacking those hydrophobic regions responsible for aggregation.

  10. Inhibition and Larvicidal Activity of Phenylpropanoids from Piper sarmentosum on Acetylcholinesterase against Mosquito Vectors and Their Binding Mode of Interaction.

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    Arshia Hematpoor

    Full Text Available Aedes aegypti, Aedes albopictus and Culex quinquefasciatus are vectors of dengue fever and West Nile virus diseases. This study was conducted to determine the toxicity, mechanism of action and the binding interaction of three active phenylpropanoids from Piper sarmentosum (Piperaceae toward late 3rd or early 4th larvae of above vectors. A bioassay guided-fractionation on the hexane extract from the roots of Piper sarmentosum led to the isolation and identification of three active phenylpropanoids; asaricin 1, isoasarone 2 and trans-asarone 3. The current study involved evaluation of the toxicity and acetylcholinesterase (AChE inhibition of these compounds against Aedes aegypti, Aedes albopictus and Culex quinquefasciatus larvae. Asaricin 1 and isoasarone 2 were highly potent against Aedes aegypti, Aedes albopictus and Culex quinquefasciatus larvae causing up to 100% mortality at ≤ 15 μg/mL concentration. The ovicidal activity of asaricin 1, isoasarone 2 and trans-asarone 3 were evaluated through egg hatching. Asaricin 1 and isoasarone 2 showed potent ovicidal activity. Ovicidal activity for both compounds was up to 95% at 25μg/mL. Asaricin 1 and isoasarone 2 showed strong inhibition on acetylcholinesterase with relative IC50 values of 0.73 to 1.87 μg/mL respectively. These findings coupled with the high AChE inhibition may suggest that asaricin 1 and isoasarone 2 are neuron toxic compounds toward Aedes aegypti, Aedes albopictus and Culex quinquefasciatus. Further computational docking with Autodock Vina elaborates the possible interaction of asaricin 1 and isoasarone 2 with three possible binding sites of AChE which includes catalytic triads (CAS: S238, E367, H480, the peripheral sites (PAS: E72, W271 and anionic binding site (W83. The binding affinity of asaricin 1 and isoasarone 2 were relatively strong with asaricin 1 showed a higher binding affinity in the anionic pocket.

  11. Salivary gland proteome analysis reveals modulation of anopheline unique proteins in insensitive acetylcholinesterase resistant Anopheles gambiae mosquitoes.

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    Sylvie Cornelie

    Full Text Available Insensitive acetylcholinesterase resistance due to a mutation in the acetylcholinesterase (ace encoding ace-1 gene confers cross-resistance to organophosphate and carbamate insecticides in Anopheles gambiae populations from Central and West Africa. This mutation is associated with a strong genetic cost revealed through alterations of some life history traits but little is known about the physiological and behavioural changes in insects bearing the ace-1(R allele. Comparative analysis of the salivary gland contents between An. gambiae susceptible and ace-1(R resistant strains was carried out to charaterize factors that could be involved in modifications of blood meal process, trophic behaviour or pathogen interaction in the insecticide-resistant mosquitoes. Differential analysis of the salivary gland protein profiles revealed differences in abundance for several proteins, two of them showing major differences between the two strains. These two proteins identified as saglin and TRIO are salivary gland-1 related proteins, a family unique to anopheline mosquitoes, one of them playing a crucial role in salivary gland invasion by Plasmodium falciparum sporozoites. Differential expression of two other proteins previously identified in the Anopheles sialome was also observed. The differentially regulated proteins are involved in pathogen invasion, blood feeding process, and protection against oxidation, relevant steps in the outcome of malaria infection. Further functional studies and insect behaviour experiments would confirm the impact of the modification of the sialome composition on blood feeding and pathogen transmission abilities of the resistant mosquitoes. The data supports the hypothesis of alterations linked to insecticide resistance in the biology of the primary vector of human malaria in Africa.

  12. Hydrogen peroxide modifies both activity and isoforms of acetylcholinesterase in human neuroblastoma SH-SY5Y cells

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    Alba Garcimartín

    2017-08-01

    Full Text Available The involvement of cholinergic system and the reactive oxygen species (ROS in the pathogenesis of some degenerative diseases has been widely reported; however, the specific impact of hydrogen peroxide (H2O2 on the acetylcholinesterase (AChE activity as well as AChE isoform levels has not been clearly established. Hence, the purpose of present study is to clarify whether H2O2 alters these parameters.Human neuroblastoma SH-SY5Y cells were treated with H2O2 (1–1000 µM for 24 h and AChE activity and AChE and cytochrome c levels were evaluated. AChE activity was strongly increased from 1 µM to 1000 µM of H2O2. The results of the kinetic study showed that H2O2 affected Vmax but not Km; and also that H2O2 changed the sigmoid kinetic observed in control samples to hyperbolic kinetic. Thus, results suggest that H2O2 acts as an allosteric activators. In addition, H2O2, (100–1000 µM reduced the total AChE content and modified its isoform profile (mainly 50-, 70-, and 132-kDa·H2O2 from 100 µM to 1000 µM induced cytochrome c release confirming cell death by apoptosis. All these results together suggest: a the involvement of oxidative stress in the imbalance of AChE; and b treatment with antioxidant agents may be a suitable strategy to protect cholinergic system alterations promoted by oxidative stress. Keywords: Acetylcholinesterase, Hydrogen peroxide, Alternative splicing, Cell culture, Cell death

  13. Polypyridylruthenium(II complexes exert in vitro and in vivo nematocidal activity and show significant inhibition of parasite acetylcholinesterases

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    Madhu Sundaraneedi

    2018-04-01

    Full Text Available Over 4.5 billion people are at risk of infection with soil transmitted helminths and there are concerns about the development of resistance to the handful of frontline nematocides in endemic populations. We investigated the anti-nematode efficacy of a series of polypyridylruthenium(II complexes and showed they were active against L3 and adult stages of Trichuris muris, the rodent homologue of the causative agent of human trichuriasis, T. trichiura. One of the compounds, Rubb12-mono, which was among the most potent in its ability to kill L3 (IC50 = 3.1 ± 0.4 μM and adult (IC50 = 5.2 ± 0.3 μM stage worms was assessed for efficacy in a mouse model of trichuriasis by administering 3 consecutive daily oral doses of the drug 3 weeks post infection with the murine whipworm Trichuris muris. Mice treated with Rubb12-mono showed an average 66% reduction (P = 0.015 in faecal egg count over two independent trials. The drugs partially exerted their activity through inhibition of acetylcholinesterases, as worms treated in vitro and in vivo showed significant decreases in the activity of this class of enzymes. Our data show that ruthenium complexes are effective against T. muris, a model gastro-intestinal nematode and soil-transmitted helminth. Further, knowledge of the target of ruthenium drugs can facilitate modification of current compounds to identify analogues which are even more effective and selective against Trichuris and other helminths of human and veterinary importance. Keywords: Acetylcholinesterase, Trichuris muris, Ruthenium complex, Anthelmintic

  14. Inhibition and Larvicidal Activity of Phenylpropanoids from Piper sarmentosum on Acetylcholinesterase against Mosquito Vectors and Their Binding Mode of Interaction.

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    Hematpoor, Arshia; Liew, Sook Yee; Chong, Wei Lim; Azirun, Mohd Sofian; Lee, Vannajan Sanghiran; Awang, Khalijah

    2016-01-01

    Aedes aegypti, Aedes albopictus and Culex quinquefasciatus are vectors of dengue fever and West Nile virus diseases. This study was conducted to determine the toxicity, mechanism of action and the binding interaction of three active phenylpropanoids from Piper sarmentosum (Piperaceae) toward late 3rd or early 4th larvae of above vectors. A bioassay guided-fractionation on the hexane extract from the roots of Piper sarmentosum led to the isolation and identification of three active phenylpropanoids; asaricin 1, isoasarone 2 and trans-asarone 3. The current study involved evaluation of the toxicity and acetylcholinesterase (AChE) inhibition of these compounds against Aedes aegypti, Aedes albopictus and Culex quinquefasciatus larvae. Asaricin 1 and isoasarone 2 were highly potent against Aedes aegypti, Aedes albopictus and Culex quinquefasciatus larvae causing up to 100% mortality at ≤ 15 μg/mL concentration. The ovicidal activity of asaricin 1, isoasarone 2 and trans-asarone 3 were evaluated through egg hatching. Asaricin 1 and isoasarone 2 showed potent ovicidal activity. Ovicidal activity for both compounds was up to 95% at 25μg/mL. Asaricin 1 and isoasarone 2 showed strong inhibition on acetylcholinesterase with relative IC50 values of 0.73 to 1.87 μg/mL respectively. These findings coupled with the high AChE inhibition may suggest that asaricin 1 and isoasarone 2 are neuron toxic compounds toward Aedes aegypti, Aedes albopictus and Culex quinquefasciatus. Further computational docking with Autodock Vina elaborates the possible interaction of asaricin 1 and isoasarone 2 with three possible binding sites of AChE which includes catalytic triads (CAS: S238, E367, H480), the peripheral sites (PAS: E72, W271) and anionic binding site (W83). The binding affinity of asaricin 1 and isoasarone 2 were relatively strong with asaricin 1 showed a higher binding affinity in the anionic pocket.

  15. Comparative study of acetylcholinesterase and glutathione S-transferase activities of closely related cave and surface Asellus aquaticus (Isopoda: Crustacea.

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    Anita Jemec

    Full Text Available The freshwater isopod crustacean Asellus aquaticus has recently been developed as an emerging invertebrate cave model for studying evolutionary and developmental biology. Mostly morphological and genetic differences between cave and surface A. aquaticus populations have been described up to now, while scarce data are available on other aspects, including physiology. The purpose of this study was to advance our understanding of the physiological differences between cave A. aquaticus and its surface-dwelling counterparts. We sampled two surface populations from the surface section of the sinking Pivka River (central Slovenia, Europe, i.e. locality Pivka Polje, and locality Planina Polje, and one cave population from the subterranean section of the sinking Pivka River, i.e. locality Planina Cave. Animals were sampled in spring, summer and autumn. We measured the activities of acetylcholinesterase (AChE and glutathione S-transferase (GST in individuals snap-frozen in the field immediately after collection. Acetylcholinesterase is likely related to animals' locomotor activity, while GST activity is related to the metabolic activity of an organism. Our study shows significantly lower AChE and GST activities in the cave population in comparison to both surface A. aquaticus populations. This confirms the assumption that cave A. aquaticus have lower locomotor and metabolic activity than surface A. aquaticus in their respective natural environments. In surface A. aquaticus populations, seasonal fluctuations in GST activity were observed, while these were less pronounced in individuals from the more stable cave environment. On the other hand, AChE activity was generally season-independent in all populations. To our knowledge, this is the first study of its kind conducted in A. aquaticus. Our results show that among closely related cave and surface A. aquaticus populations also physiological differences are present besides the morphological and genetic

  16. Screen-printed electrode modified with carbon black and chitosan: a novel platform for acetylcholinesterase biosensor development.

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    Talarico, Daria; Arduini, Fabiana; Amine, Aziz; Cacciotti, Ilaria; Moscone, Danila; Palleschi, Giuseppe

    2016-10-01

    We report a screen-printed electrode (SPE) modified with a dispersion of carbon black (CB) and chitosan by drop casting. A cyclic voltammetry technique towards ferricyanide, caffeic acid, hydroquinone, and thiocholine was performed and an improvement of the electrochemical response with respect to bare SPE as well as SPE modified only with chitosan was observed. The possibility to detect thiocholine at a low applied potential with high sensitivity was exploited and an acetylcholinesterase (AChE) biosensor was developed. A dispersion of CB, chitosan, and AChE was used to fabricate this biosensor in one step by drop casting. The enzymatic activity of the immobilized AChE was determined measuring the enzymatic product thiocholine at +300 mV. Owing to the capability of organophosphorus pesticides to inhibit AChE, this biosensor was used to detect these pollutants, and paraoxon was taken as model compound. The enzyme inhibition was linearly related to the concentration of paraoxon up to 0.5 μg L(-1), and a low detection limit equal to 0.05 μg L(-1) (calculated as 10% of inhibition) was achieved. This biosensor was challenged for paraoxon detection in drinking waters with satisfactory recovery values. The use of AChE embedded in a dispersion of CB and chitosan allowed an easy and fast production of a sensitive biosensor suitable for paraoxon detection in drinking waters at legal limit levels. Graphical Abstract Biosensors based on screen-printed electrodes modified with Acetylcholinesterase, Carbon Black, and Chitosan for organophosphorus pesticide detection.

  17. Acetylcholinesterase immobilization and characterization, and comparison of the activity of the porous silicon-immobilized enzyme with its free counterpart.

    Science.gov (United States)

    Saleem, Muhammad; Rafiq, Muhammad; Seo, Sung-Yum; Lee, Ki Hwan

    2016-02-02

    A successful prescription is presented for acetylcholinesterase physically adsorbed on to a mesoporous silicon surface, with a promising hydrolytic response towards acetylthiocholine iodide. The catalytic behaviour of the immobilized enzyme was assessed by spectrophotometric bioassay using neostigmine methyl sulfate as a standard acetycholinesterase inhibitor. The surface modification was studied through field emission SEM, Fourier transform IR spectroscopy, energy-dispersive X-ray spectroscopy, cathode luminescence and X-ray photoelectron spectroscopy analysis, photoluminescence measurement and spectrophotometric bioassay. The porous silicon-immobilized enzyme not only yielded greater enzyme stability, but also significantly improved the native photoluminescence at room temperature of the bare porous silicon architecture. The results indicated the promising catalytic behaviour of immobilized enzyme compared with that of its free counterpart, with a greater stability, and that it aided reusability and easy separation from the reaction mixture. The porous silicon-immobilized enzyme was found to retain 50% of its activity, promising thermal stability up to 90°C, reusability for up to three cycles, pH stability over a broad pH of 4-9 and a shelf-life of 44 days, with an optimal hydrolytic response towards acetylthiocholine iodide at variable drug concentrations. On the basis of these findings, it was believed that the porous silicon-immobilized enzyme could be exploited as a reusable biocatalyst and for screening of acetylcholinesterase inhibitors from crude plant extracts and synthesized organic compounds. Moreover, the immobilized enzyme could offer a great deal as a viable biocatalyst in bioprocessing for the chemical and pharmaceutical industries, and bioremediation to enhance productivity and robustness. © 2016 Authors.

  18. Parasite distribution and early-stage encephalitis in Sarcocystis calchasi infections in domestic pigeons (Columba livia f. domestica).

    Science.gov (United States)

    Maier, Kristina; Olias, Philipp; Enderlein, Dirk; Klopfleisch, Robert; Mayr, Sylvia L; Gruber, Achim D; Lierz, Michael

    2015-01-01

    Pigeon protozoal encephalitis is a biphasic, neurologic disease of domestic pigeons (Columba livia f. domestica) caused by the apicomplexan parasite Sarcocystis calchasi. Despite severe inflammatory lesions of the brain, associated parasitic stages have only rarely been identified and the cause of the lesions is still unclear. The aim of this study was therefore to characterize the tissue distribution of S. calchasi within pigeons between the two clinical phases and during the occurrence of neurological signs. For this purpose, a semi-quantitative real-time polymerase chain reaction (PCR) was developed. Forty-five domestic pigeons were infected orally (via a cannula into the crop) with 200 S. calchasi sporocysts and euthanized in groups of three pigeons at intervals of 2 to 10 days over a period of 61 days. Tissue samples including brain and skeletal muscle were examined by histology, immunohistochemistry, and PCR. Schizonts were detected in the liver of one pigeon at day 10 post infection. A mild encephalitis was detected at day 20 post infection, around 4 weeks before the onset of neurological signs. At the same time, immature sarcocysts were present in the skeletal muscle. In seven pigeons a few sarcocysts were identified in the brain, but not associated with any lesion. These results suggest that the encephalitis is induced at a very early stage of the S. calchasi lifecycle rather than in the chronic phase of pigeon protozoal encephalitis. Despite the increasing severity of lesions in the central nervous system, the amount of sarcocysts did not increase. This supports the hypothesis of a delayed-type hypersensitivity response as the cause of the encephalitis. The study also demonstrated that S. calchasi DNA is detectable in tissues negative by histological methods, indicating a higher sensitivity of the real-time PCR.

  19. Putative adult neurogenesis in two domestic pigeon breeds (Columba livia domestica): racing homer versus utility carneau pigeons.

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    Mazengenya, Pedzisai; Bhagwandin, Adhil; Nkomozepi, Pilani; Manger, Paul R; Ihunwo, Amadi O

    2017-07-01

    Generation of neurons in the brains of adult birds has been studied extensively in the telencephalon of song birds and few studies are reported on the distribution of PCNA and DCX in the telencephalon of adult non-song learning birds. We report here on adult neurogenesis throughout the brains of two breeds of adult domestic pigeons (Columba livia domestica), the racing homer and utility carneau using endogenous immunohistochemical markers proliferating cell nuclear antigen (PCNA) for proliferating cells and doublecortin (DCX) for immature and migrating neurons. The distribution of PCNA and DCX immunoreactivity was very similar in both pigeon breeds with only a few minor differences. In both pigeons, PCNA and DCX immunoreactivity was observed in the olfactory bulbs, walls of the lateral ventricle, telencephalic subdivisions of the pallium and subpallium, diencephalon, mesencephalon and cerebellum. Generally, the olfactory bulbs and telencephalon had more PCNA and DCX cells than other regions. Two proliferative hotspots were evident in the dorsal and ventral poles of the lateral ventricles. PCNA- and DCX-immunoreactive cells migrated radially from the walls of the lateral ventricle into the parenchyma. In most telencephalic regions, the density of PCNA- and DCX-immunoreactive cells increased from rostral to caudal, except in the mesopallium where the density decreased from rostral to middle levels and then increased caudally. DCX immunoreactivity was more intense in fibres than in cell bodies and DCX-immunoreactive cells included small granular cells, fusiform bipolar cells, large round and or polygonal multipolar cells. The similarity in the distribution of proliferating cells and new neurons in the telencephalon of the two breeds of pigeons may suggest that adult neurogenesis is a conserved trait as an ecological adaptation irrespective of body size.

  20. Putative adult neurogenesis in two domestic pigeon breeds (Columba livia domestica: racing homer versus utility carneau pigeons

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    Pedzisai Mazengenya

    2017-01-01

    Full Text Available Generation of neurons in the brains of adult birds has been studied extensively in the telencephalon of song birds and few studies are reported on the distribution of PCNA and DCX in the telencephalon of adult non-song learning birds. We report here on adult neurogenesis throughout the brains of two breeds of adult domestic pigeons (Columba livia domestica, the racing homer and utility carneau using endogenous immunohistochemical markers proliferating cell nuclear antigen (PCNA for proliferating cells and doublecortin (DCX for immature and migrating neurons. The distribution of PCNA and DCX immunoreactivity was very similar in both pigeon breeds with only a few minor differences. In both pigeons, PCNA and DCX immunoreactivity was observed in the olfactory bulbs, walls of the lateral ventricle, telencephalic subdivisions of the pallium and subpallium, diencephalon, mesencephalon and cerebellum. Generally, the olfactory bulbs and telencephalon had more PCNA and DCX cells than other regions. Two proliferative hotspots were evident in the dorsal and ventral poles of the lateral ventricles. PCNA- and DCX-immunoreactive cells migrated radially from the walls of the lateral ventricle into the parenchyma. In most telencephalic regions, the density of PCNA- and DCX-immunoreactive cells increased from rostral to caudal, except in the mesopallium where the density decreased from rostral to middle levels and then increased caudally. DCX immunoreactivity was more intense in fibres than in cell bodies and DCX-immunoreactive cells included small granular cells, fusiform bipolar cells, large round and or polygonal multipolar cells. The similarity in the distribution of proliferating cells and new neurons in the telencephalon of the two breeds of pigeons may suggest that adult neurogenesis is a conserved trait as an ecological adaptation irrespective of body size.