Detoxification enzymes

Le mot molybdène du grec ancien μόλυβδος  (plomb) désigne un oligoélément minéral stocké dans le foie et  les reins. C’est un cofacteur de plusieurs métallo-enzymes, intervenant dans le métabolisme de fer, de la synthèse de protéines, d'enzymes et des purines qui entrent dans la composition de l'ADN, dans l’élimination de l'acide urique, la solidité des dents, la capacité érectile, la croissance osseuse. Il prévient  de certaines tumeurs au niveau de l'œsophage, de la carie dentaire. C’est un antioxydant  et un  détoxiquant. Il interagit avec le fer, le zinc et le cuivre. Il est surtout présent dans les produits animaux tels que : les abats, le foie, les rognons,  les produits laitiers; les fruits de mer. Il se trouve également dans les végétaux :  graines, noix, noisettes, amandes ; céréales : germes de blé, sarrasin, avoine, sorgho ; les légumineuses et légumes comme : pois sec,  pois chiche,  petit pois, lentilles, haricots, céleri rave, radis, betterave, haricots, courgette, céleri-rave, ail, tomate et épinard ; les fruits à coque : noix,  amandes et noisettes ; les fruits tropicaux : papaye, noix de coco, goyave. La carence et l’excès peuvent être remédiés par des plantes.

Background: The whitefly Trialeurodes vaporariorum is an economically important crop pest in temperate regions that has developed resistance to most classes of insecticides. However, the molecular mechanisms underlying resistance have not been characterised and, to date, progress has been hampered by a lack of nucleotide sequence data for this species. Here, we use pyrosequencing on the Roche 454-FLX platform to produce a substantial and annotated EST dataset. This 'unigene set' will form a critical reference point for quantitation of over-expressed messages via digital transcriptomics. Results: Pyrosequencing produced around a million sequencing reads that assembled into 54,748 contigs, with an average length of 965 bp, representing a dramatic expansion of existing cDNA sequences available for T. vaporariorum (only 43 entries in GenBank at the time of this publication). BLAST searching of non-redundant databases returned 20,333 significant matches and those gene families potentially encoding gene products involved in insecticide resistance were manually curated and annotated. These include, enzymes potentially involved in the detoxification of xenobiotics and those encoding the targets of the major chemical classes of insecticides. A total of 57 P450s, 17 GSTs and 27 CCEs were identified along with 30 contigs encoding the target proteins of six different insecticide classes. Conclusion: Here, we have developed new transcriptomic resources for T. vaporariorum. These include a substantial and annotated EST dataset that will serve the community studying this important crop pest and will elucidate further the molecular mechanisms underlying insecticide resistance.

The toxicity of chlorpyrifos to three Iranian populations of two-spotted spider mite, collected from Isfahan (ISR), Yazd (Yz) and Guilan (GUS2) Provinces were surveyed using the residual contact vial bioassay. The bioassay results showed that resistance ratios of ISR and Yz populations were 176.90 and 9.78 fold compared to the GUS2 population, respectively. Determination of esterase and glutathione-S-transferase activity and their kinetic parameters showed that ISR population had the highest specific activity and specificity constant among the studied populations. Besides, the content of mixed function oxidases in ISR population was the highest. However, synergistic effects of Piperonyl Butoxide, Diethyl Maleate and Triphenyl Phosphate showed that metabolic enzymes did not play an important role in resistance to chlorpyrifos in ISR and Yz populations and enhanced activity of esterase, glutathione-S-transferase and content of mixed function oxidases in these populations were probably due to resistance to some other acaricides. To determine the role of acetylcholinesterase insensitivity in resistance mechanisms, kinetic parameters and inhibitory effect of chlorpyrifos-oxon on this enzyme were investigated. The K m value of acetylcholinesterase was determined as 0.036, 0.04, and 0.050 mM using acetylthiocholine iodide for GUS2, Yz, and ISR populations, respectively. In addition, the insensitivity ratios of chlorpyrifos-oxon on acetylcholinesterase activity were estimated at 23.30 and 2.96 for ISR and Yz populations, respectively. These results confirmed amino acid substitutions in active site of this enzyme and also indicated that resistant population possed qualitatively altered AChE.

It is known that pesticide resistance is either caused by altered target sites, such as
amino acid changes in voltage-gated sodium channels in some insects (Soderland & Knipple,
2003) or by increased amounts of detoxifying enzymes. The most prevalent resistance
mechanism is the upregulation of these detoxification enzymes. Nilaparvata lugens, is one of
the four principal (major) detoxifying enzymes included in the IR in BPH (Lu et al., 2021a),
which are:
• Cytochrome P450 monooxygenase (P450s),
• Glutathione-S-Transferase (GSTs)
• Carboxylesterases (CarEs), and
• UDP-glycosyltransferases (UGTs)

DESCUBIERTO ­ ¡La verdad absoluta sobre la desintoxicación y la pérdida de peso!

La desintoxicación es algo muy importante para el cuerpo, pero es algo que no se comprende bien.

Hace siglos, los maestros de la salud en Oriente comprendieron la importancia de equilibrar y desintoxicar el cuerpo. Es algo que la medicina occidental apenas comienza a comprender

We assessed whether exposure to environmentally-relevant mixtures of four organophosphorus insecticides (OPs) exerted adverse effects on honey bees. Adult and worker bees were orally exposed for five days under laboratory conditions to mixtures of four insecticides, diazinon, malathion, profenofos and chlorpyrifos at two concentrations. Concentration in the mixtures tested were equivalent to the median and 95th centile concentrations of the OPs in honey, as reported in the literature. Effects on survival, behavior, activity of acetylcholinesterase (AChE), and expression of genes important in detoxification of xenobiotics and immune response were examined. Survival of worker bees was not affected by exposure to median or 95th centile concentrations of the OPs. Activity of AChE was significantly greater in worker bees exposed to the 95th centile concentration mixture of OPs compared to the median concentration mixture. Expression of genes involved in detoxification of xenobiotics was not affected by treatment, but the abundance of transcripts of the antimicrobial peptide hymenoptaecin was significantly greater in worker honey bees exposed to the median concentration mixture. Results suggest that short-term exposure to environmentally relevant concentrations of a mixture of OPs do not adversely affect worker honey bees.

Due to the accumulation of heavy metal compounds formed in plants for food that lead to human toxicity, and probably high incidence of diseases, ,detoxification of Cadmium and Lead as heavy metals in Oryza sativa rice by treated Pistachio Shell (PS) and Eggshells(ES) as two available and novel agro and food bio-wastes has been studied. The originally native rice paddies area was used for years under conventional management for more than 30 years, and the soil was sampled twice (14 and 180 days after incorporating of bio-adsorbents : Pistachio Shell and Eggshell, in 10 hectors in each studied sites). Current assessment was designed principally to appraise the capacity of two bio-adsorbents from food waste sector for the bio-removal of cadmium and lead ions from contaminated soil of paddy rice samples after the determination of these metals in such rice samples in different agricultural sites. Heavy metals in 900 treated and untreated rice samples were analyzed using Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). Results were determined as mean ± Standard Deviation of dry weight from three replicates in each test. The mean contents of Cadmium and Lead in most of untreated samples were above maximum level which is recommended by FAO/WHO Expert Committee on Food Additives (JECFA). Lead and Cadmium concentrations in all treated samples by bio-wastes decreased significantly (p<0.001) and turned into lower than the maximum permissible limits set by International Committees and also Iranian National Standardization Organization. The findings of present survey indicate that Food /Agro wastes can be utilized in healing of contaminated food stuffs specially rice containing heavy metal ions.

The context is the post-industrial regeneration of public spaces, the transformation of contaminated areas in urban park, through strategies of both environmental sustainability and eco-compatibility, both creative and cultural landscape enhancement. In the framework of ‘Spine 4&#39; park in Turin, named Aurelio Peccei, an integrated and innovative competition, design, evaluation, performance analysis, engineering and execution, dedicated to public art has implemented, which led to the realization of ten works. The evaluation of performance and cost in the life cycle is directed to develop research and testing to the first italian outdoor art and design works in cements nanostructured UHPC (Ultra High Performance Concrete). / Il contesto e la rigenerazione degli spazi pubblici post-industriali, la trasformazione di aree inquinate in parco urbano, attraverso strategie sia di sostenibilita ambientale ed eco-compatibilita che di valorizzazione paesaggistica creativa e culturale. Nell&#...

The toxicity of acrolein, an a,b-unsaturated aldehyde, is due to its soft electrophilic nature and primarily involves the adduction of protein thiols. The thiol glutathione (GSH) forms the first line of defense against acrolein. The present study confirms that acrolein added to isolated rat liver microsomes can increase microsomal GSH transferase (MGST) activity 2e3 fold, which can be seen as a direct adaptive increase in the protection against acrolein. At a relatively high exposure level, acrolein appeared to inhibit MGST. The activation is due to adduction of thiol groups, and the inactivation probably involves adduction of amino groups in the enzyme by acrolein. The preference of acrolein to react with thiol groups over amino groups can explain why the enzyme is activated at a low exposure level and inhibited at a high exposure level of acrolein. These opposite forms of direct adaptation on the level of enzyme activity further narrow the thin line between survival and promotion of cell death, governed by the level of exposure.

People interested in the research are advised to contact the author for the final version of the publication, or visit the DOI to the publisher's website. • The final author version and the galley proof are versions of the publication after peer review. • The final published version features the final layout of the paper including the volume, issue and page numbers. Link to publication General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal. If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the "Taverne" license above, please follow below link for the End User Agreement:

The fungicide carbendazim (CBM) has been applied all around the world but its potential adverse effects other than its recognized activity as endocrine disruptor in non target organisms have been scarcely studied. The aims of this work were (1) to use a battery of biomarkers that can reflect potential negative effects such as oxidative stress, genotoxicity, neurotoxicity or altered immune response; and (2) to examine biomarkers of detoxification by analyzing the gene expression of cytochrome P4501A1 (CYP1A1) and the multi-xenobiotic resistance protein P-glycoprotein (P-gp) in the freshwater fish Jenynsia multidentata exposed to environmentally relevant concentrations of CBM during 24 h. Fish exposed to 5 μg/L showed inhibition of GST activity and an increase of TBARs contents in gills, the organ of direct contact with waterborne contaminants. Genotoxicitymeasured in peripheral blood-was evidenced by the increases of micronuclei frequency when fish were exposed to 5, 10 and 100 μg/L CBM and of nuclear abnormalities (NA) frequency at 0.05, 0.5, 5, 10 and 100 μg/L CBM. The expression inhibition of interleukin (IL-1β) and tumor necrosis factor a (TNF-α) at 10, and 5 and 10 μg/L CBM, respectively, indicated an altered immune response. The expression of CYP1A1 was down regulated in liver at 10 μg/L and of P-gp at 5 μg/L CBM, indicating a possible slow on CBM metabolization. On the other hand, in gills CYP1A1 decreased at 5 and 10 μg/L while P-gp was induced at 5 and 100 μg/L CBM. Overall, most of these significant effects were detected below 10 μg/L CBM, in a range of realistic concentrations in aquatic ecosystems worldwide.

Despite the crucial role of highly social bees (Hymenoptera, Apidae) in crop production, agricultural practices that embrace the use of chemicals for pest control put these important pollinators at risk. Most studies that have evaluated the expression of enzymatic routes involved in xenobiotic removal in these species, including pesticides, have been carried out in the honey bee, Apis mellifera. These results have been used to gure out the response of non-Apis bees to pesticides, but recent works suggest that such response may be speciesspeci c. We tested this possibility by evaluating the glutathione S-transferase (GST), cytochrome P 450 , acetylcholinesterase (AChE) and α-, β-and ρ-NPA esterases in foragers both of the stingless bee Scaptotrigona mexicana and of A. mellifera. Foragers from colonies of both species located in two sites in an agricultural landscape in Chiapas, Mexico were collected every four months for a year and enzymatic expression was measured. Scaptotrigona mexicana showed a higher enzymatic activity and AChE inhibition than A. mellifera in both sites. Neither site nor time affected statistically the expression of enzymes in each species. Our results suggest that A. mellifera cannot be used as a surrogate for other species, at least for S. mexicana. The higher enzymatic activities in S. mexicana related to A. mellifera can be explained by the presence of organochlorines and organophosphates in the study area, as revealed in previous studies, and the shorter y range of S. mexicana.

Glutathione transferases (GSTs) form a superfamily of multifunctional proteins with essential roles in cellular detoxification processes and endogenous metabolism. The distribution of fungal-specific class A GSTs was investigated in saprotrophic fungi revealing a recent diversification within this class. Biochemical characterization of eight GSTFuA isoforms from Phanerochaete chrysosporium and Coprinus cinereus demonstrated functional diversity in saprotrophic fungi. The threedimensional structures of three P. chrysosporium isoforms feature structural differences explaining the functional diversity of these enzymes. Competition experiments between fluorescent probes, and various molecules, showed that these GSTs function as ligandins with various small aromatic compounds, derived from lignin degradation or not, at a L-site overlapping the glutathione binding pocket. By combining genomic data with structural and biochemical determinations, we propose that this class of GST has evolved in response to environmental constraints induced by wood chemistry.

People interested in the research are advised to contact the author for the final version of the publication, or visit the DOI to the publisher's website. • The final author version and the galley proof are versions of the publication after peer review. • The final published version features the final layout of the paper including the volume, issue and page numbers. Link to publication General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal. If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the "Taverne" license above, please follow below link for the End User Agreement:

Status and Biochemical Resistance Of Crocidolomia pavonana (F.) (Lepidoptera: Crambidae) to Organophosphate Insecticide and Its Sensitivity to Botanical Insecticide. An examination of insect resistance was determined by several steps, i.e. standard sensitivity, resistance diagnosis, and determination of resistance level. Each phase was tested with feeding and residue contact methods at glass tube. Resistance ratio (RR) was determined by comparing LC 50 value of field population with standard population. Field population of C. pavonana was classified resistant if it had RR ≥ 4. Biochemistry analysis of resistance was conducted to population of C. pavonana showing resistance to prophenophos insecticide. The activity analysis of acetylcholine esterase (ACHE), esterase, and Glutation Stransferase was done with spectrophotometer method. Insect which are resistant to prophenophos insecticide was tested for its sensitivity to Barringtonia asiatica seed extract. Result indicated that C. pavonana population from Pengalengan showed resistance to prophenophos synthetic insecticide. Using contact test, the highest resistance ratio value was 4.04, while by feeding assay the RR was 2.78. The study on biochemical resistance mechanisms of each field population of C. pavonana showed various activities of enzymatic detoxification. This could be due to the difference in the kind of insecticides exposed to each field population of C. pavonana. Since RR value from the contact test was higher than that of the feeding test, the resistance development of C. pavonana to synthetic insecticides was probably caused by physiological and biochemical changes in insect cuticle rather than the activity of detoxification enzyme. Methanolic seed extract of B. asiatica can be used as an alternative of resistance management of C. pavonana to prophenophos synthetic insecticide.

This research aims to study 1) Maternal effect on the inheritance of yield components of Manado Yellow Use of Insecticides is one way that is very often made vegetable farmersto cope with pest and plant diseases. However, the use of insecticides may give negative effects for consumers of food containing residues of these poisons.Methodology of research is survey and laboratory. Collecting data in the survey was an interview of 20 vegetable farmers, each 10 from Modoinding and 10 from Rurukan that determined purposively. Laboratory data obtainedwith the ways to identify residues in the cabbage and tomatoes from Modoinding and Rurukan , using Gas Chromatography (GC) according to the analysis method of Pusat Pengujian Obat dan Makanan.Vegetable farmers in Modoinding and Rurukan often apply the insecticide organophospate and pyretroid on cabbage and tomato plants. The residues level in ccabbage and tomato from Modoinding and Rurukan far exceed the value of Acceptable Daily Intake of profonefos allowed.

ABSTRAK Aedes aegypti (Linnaeus) (Diptera: Culicidae) merupakan serangga vektor utama penyakit demam berdarah. Di Indonesia, cara yang selama ini terbukti efektif untuk mengendalikan Ae. aegypti adalah dengan menggunakan insektisida terutama piretroid, seperti permetrin. Meskipun permetrin telah digunakan sejak tahun 1980an, informasi mengenai status dan perkembangan resistensi Ae. aegypti terhadap permetrin masih terbatas dan perlu dilakukan pembaharuan data. Penelitian ini dilakukan menggunakan metode uji standar WHO bertujuan untuk mengetahui status dan perubahan tingkat resistensi Ae. aegypti strain lapangan, yaitu Palu, Makassar, Bandung, Bogor, dan strain rentan (VCRU) yang diseleksi dengan permetrin selama lima generasi. Hasil penelitian menunjukkan bahwa semua strain lapangan telah resisten terhadap permetrin dengan tingkat resistensi tinggi (nilai RR 50 yang berkisar antara 10,5 dan 25,7). Strain Bandung memperlihatkan RR 50 tertinggi (25,7) dan terendah pada strain Makassar (10,5). Lima strain yang diseleksi mengalami kenaikan tingkat resistensi 5-18 kali lebih tinggi pada F5 dibandingkan dengan parentalnya. Pengetahuan mengenai status resistensi pada area tertentu ditambah dengan pemahaman tentang kecepatan terjadinya resistensi, dapat digunakan untuk merancang program pengendalian vektor yang lebih baik.

People interested in the research are advised to contact the author for the final version of the publication, or visit the DOI to the publisher's website. • The final author version and the galley proof are versions of the publication after peer review. • The final published version features the final layout of the paper including the volume, issue and page numbers. Link to publication General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal. If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the "Taverne" license above, please follow below link for the End User Agreement:

Functional relationship of a-glutathione S-t ransferase and gl utat h ione S-t ra nsferase activity in machine-preserved non-heart-beating donor kidneys Abstract a-Glutathione S-transferase (a-GST) is a biochemical parameter used to estimate the amount of proximal tubule damage to a kidney. In normal clinical practice, the concentration of a-GST in urine is determined by a rather time-consuming immunochemical test, the enzyme-linked immunosorbent assay (ELISA). Kidneys from nonheart-beating (NHB) donors are perfused prior to transplantation. The determination of a-GST concentration in the perfusate to monitor damage is also done by means of an ELISA test. However, because this is a time-consuming method, it would be helpful to find a parameter proportional to GST concentrations that would be available within minutes. We therefore compared the method of determining a-GST concentration via ELISA with that of determining the enzymatic activity of GST, which is much faster (results available within 10 min). The comparison was made using 150 preserved kidneys that had been perfused for 6 h. The correlation was found to be very good, as indicated by the linear regression data: r = 0.954, P < 0.001. n-Glutathione S-transferase (n-GST) was also determined by means of an ELISA test, and the concentration of n-GST was compared with the enzymatic activity of the "total" GST. The precision of the enzymatic method, given by intra-and interassay variation, was 1.5% and 10.5%, respectively. Keywords GST activity. a-GST. Acute tubule necrosis. Proximal tubule damage

Heterotrigona itama is a Malaysian stingless bee species that actively reared for meliponiculture. This stingless bee is cultivated in a commercial scale for its honey production, propolis and among the greatest commercial potential as crop pollinators. However, this species has been potentially exposed to agronomic practices, among which the use of synthetic insecticides against pests. The indirect toxicity effect of the post-insecticide had affected the mortalities of H. itama especially, to the foragers. Due to that, a study has been conducted to determine the lethal concentration of 50% (LC50) and 95% (LC95) of the selected insecticides against stingless bee forager workers through residual exposure. The bioassay test was conducted to the local stingless bee H. itama at Agricultural Research Station, Tenom. Four commonly used insecticides in crop protection; Deltamethrin, Chlorpyrifos, Cypermethrin and Malathion were tested at five concentrations that diluted with 500 ml of distilled water in three replications for each insecticide. Lethal concentrations (LC50 and LC95) were obtained from probit analysis after 1-hour dry residues exposure and 24-hour mortality observation. The result shows that; all four tested insecticides were harmful to H. itama through dry residue. Deltamethrin shows the higher value of LC50 (1.256 ml) and LC95 (3.582ml) that make it less toxic to the H. itama than cypermethrin, malathion, and chlorpyrifos, however, as the concentration gets higher it becomes more toxic.

Abstract One factor proposed to explain, wholly or in part, losses of honey bee colonies is wide-spread application of agrochemicals. Concentrations of 14 organophosphorous insecticides (OPs) were measured in bees and hive matrices including honey and bee bread to assess their potential hazard to honey bees. Samples were collected from 7 randomly selected colonies in Saskatchewan during summer of 2013. LC/MS-MS was used to identify and quantify individual OPs by use of a modified QuickEasyCheapEffectiveRuggedSafe (QUECHERS) method. Diazinon, dimethoate and chlorpyrifos-oxon were the only OPs detected in honey with mean concentrations of 0.25, 1.5 and 0.21 ng/g, wet mass (wm) respectively. Fenamiphos, chlorpyrifos and chlorpyrifos methyl were the only OPs detected in bee bread, with mean concentrations of 0.3, 2.7 and 1.6 x101 ng/g, wm, respectively, while, ethoprop, malathion and dichlorvos were the only OPs detected in bees with mean concentrations of 1.4, 3.7 and 8.9 x102 ng /g, w...

Satu buah glikosida oleanan berhasil diisolasi dari biji Barringtonia asiatica . Struktur  senyawa ini ditentukan oleh 1H- dan 13C-NMR satu dan dua dimensi serta oleh perbandingan langsung dengan standar. Senyawa ini menunjukkan aktivitas insektisida yang paling tinggi terhadap Crocidolomia pavonana. Dari penelitian ini diketahui bahwa biji B. asiatica mengandung senyawa insektisida yang paling aktif dengan nilai LC50 sebesar 290 ppm yang potensial untuk aplikasi insektisida. Kata Kunci: Barringtonia asiatica , glikosida oleanan, aktivitas insektisida, Crocidolomia pavonana

This research was conducted to know resistance level of Crocidolomia pavonana to a profenofos synthetic insecticide and to know susceptibility of C. pavonana to Azadirachta indica seed extract. The experiment using two bioassay methods, dry film method and feeding method. Field population of C. pavonana obtained from 3 villages in Lembang district i.e.Cibogo, Cikole and Kayu Ambon village. The standard insect has been maintenance more than 20 generations in the laboratory as a comparison. The result of the experiment indicated that population of C. pavonana form Cikole, Cibogo and Kayu Ambon village were resistant to profenofos insecticide with resistance ratio (RR) was low (1< RR ≤ 5). Resistance Ratio of C. pavonana from the third village were 4.69 and 4.83; 1.02 and 1.1 and 1.16 and 1.9 respectively (dry film and feeding method). The value of the resistance ratio was greater through the feeding assay compared to the contact assay. This indicated that the mechanism of C. pavonana resistance was likely to occur mainly in the digestive tract of insects. Enzime activity of the different sample population of C. pavonana indicated variation in their activity. This was indicated that every sample from different field could develop different mechanism of resistance. The toxicity study of A. indica seed extract to C. pavonana indicate that the field population of C. pavonana from the third village was susceptible to seed extract of A. indica with value RR of less than one point. Therefore seed extract of A. indica could be used as alternative control against C. pavonana which resistant to profenofos synthetic insecticide.

The completion of the sequencing of the human genome [1,2] has opened an unprecedented opportunity to develop novel molecular diagnostic and therapeutic approaches, based on a far deeper understanding of the underlying pathophysiology of many complex disorders. Genomics-based research aims to link genotype to phenotype and requires access to high quality human tissue and cell lines, which presents a series of unique challenges. The first IBC Human Tissue in Target Identification and Drug Discovery Conference (3-4 December 2001; Boston, MA, USA) specifically addressed this important topic. Target identification and drug discovery One of the major challenges facing the drug discovery community is the limitation and poor predictability of animalbased strategies. Many drugs have failed in later stages of development because the animal data were poor predictors of efficacy in the human subject. The case for using human tissue early in the discovery process was elegantly made by Michael Neubauer (Bristol Myers Squibb, Princeton, NJ, USA), who described the multiple points at which microarray technology and human tissue could be integrated to aid in target identification and validation, safety testing, compound selection and crucial decision making on parameters to advance products to clinical testing. Ways of integrating the target discovery phase with the drug discovery phase based on the creative use of human tissues, stem cells and microarray capability were emphasized by the keynote speaker,

The red imported fire ant (Solenopsis invicta) is one of the deadliest invasive ant species that threatens the world by disrupting biodiversity, important functions within a natural ecosystem, and community structure. They are responsible for huge economic losses in the infested countries every year. Synthetic insecticides, especially indoxacarb, have been broadly used to control S. invicta for many years. However, the biochemical response of S. invicta to indoxacarb remains largely undiscovered. Here, we used the sublethal doses of indoxacarb on the S. invicta collected from the eight different cities of Southern China. The alteration in the transcriptome profile of S. invicta following sublethal dosages of indoxacarb was characterized using high-throughput RNA-seq technology. We created 2 libraries, with 50.93 million and 47.44 million clean reads for indoxacarb treatment and control, respectively. A total of 2018 unigenes were regulated after insecticide treatment. Results indica...

Background: The whitefly Trialeurodes vaporariorum is an economically important crop pest in temperate regions that has developed resistance to most classes of insecticides. However, the molecular mechanisms underlying resistance have not been characterised and, to date, progress has been hampered by a lack of nucleotide sequence data for this species. Here, we use pyrosequencing on the Roche 454-FLX platform to produce a substantial and annotated EST dataset. This 'unigene set' will form a critical reference point for quantitation of over-expressed messages via digital transcriptomics. Results: Pyrosequencing produced around a million sequencing reads that assembled into 54,748 contigs, with an average length of 965 bp, representing a dramatic expansion of existing cDNA sequences available for T. vaporariorum (only 43 entries in GenBank at the time of this publication). BLAST searching of non-redundant databases returned 20,333 significant matches and those gene families potentially encoding gene products involved in insecticide resistance were manually curated and annotated. These include, enzymes potentially involved in the detoxification of xenobiotics and those encoding the targets of the major chemical classes of insecticides. A total of 57 P450s, 17 GSTs and 27 CCEs were identified along with 30 contigs encoding the target proteins of six different insecticide classes. Conclusion: Here, we have developed new transcriptomic resources for T. vaporariorum. These include a substantial and annotated EST dataset that will serve the community studying this important crop pest and will elucidate further the molecular mechanisms underlying insecticide resistance.

An examination of insect resistance was determined by several steps, i.e. standard sensitivity, resistance diagnosis, and determination of resistance level. Each phase was tested with feeding and residue contact methods at glass tube. Resistance ratio (RR) was determined by comparing LC50 value of field population with standard population. Field population of C. pavonana was classified resistant if it had RR 5 4. Biochemistry analysis of resistance was conducted to population of C. pavonana showing resistance to prophenophos insecticide. The activity analysis of acetylcholine esterase (ACHE), esterase, and Glutation Stransferase was done with spectrophotometer method. Insect which are resistant to prophenophos insecticide was tested for its sensitivity to Barringtonia asiatica seed extract. Result indicated that C. pavonana population from Pengalengan showed resistance to prophenophos synthetic insecticide. Using contact test, the highest resistance ratio value was 4.04, while by fe...

An examination of insect resistance was determined by several steps, i.e. standard sensitivity, resistance diagnosis, and determination of resistance level. Each phase was tested with feeding and residue contact methods at glass tube. Resistance ratio (RR) was determined by comparing LC50 value of field population with standard population. Field population of C. pavonana was classified resistant if it had RR 5 4. Biochemistry analysis of resistance was conducted to population of C. pavonana showing resistance to prophenophos insecticide. The activity analysis of acetylcholine esterase (ACHE), esterase, and Glutation Stransferase was done with spectrophotometer method. Insect which are resistant to prophenophos insecticide was tested for its sensitivity to Barringtonia asiatica seed extract. Result indicated that C. pavonana population from Pengalengan showed resistance to prophenophos synthetic insecticide. Using contact test, the highest resistance ratio value was 4.04, while by fe...

Background: The whitefly Trialeurodes vaporariorum is an economically important crop pest in temperate regions that has developed resistance to most classes of insecticides. However, the molecular mechanisms underlying resistance have not been characterised and, to date, progress has been hampered by a lack of nucleotide sequence data for this species. Here, we use pyrosequencing on the Roche 454-FLX platform to produce a substantial and annotated EST dataset. This 'unigene set' will form a critical reference point for quantitation of over-expressed messages via digital transcriptomics. Results: Pyrosequencing produced around a million sequencing reads that assembled into 54,748 contigs, with an average length of 965 bp, representing a dramatic expansion of existing cDNA sequences available for T. vaporariorum (only 43 entries in GenBank at the time of this publication). BLAST searching of non-redundant databases returned 20,333 significant matches and those gene families potentially encoding gene products involved in insecticide resistance were manually curated and annotated. These include, enzymes potentially involved in the detoxification of xenobiotics and those encoding the targets of the major chemical classes of insecticides. A total of 57 P450s, 17 GSTs and 27 CCEs were identified along with 30 contigs encoding the target proteins of six different insecticide classes. Conclusion: Here, we have developed new transcriptomic resources for T. vaporariorum. These include a substantial and annotated EST dataset that will serve the community studying this important crop pest and will elucidate further the molecular mechanisms underlying insecticide resistance.

The present study was designed to evaluate the susceptibility of Culex quinquefasciatuns against malathion in Sargodha district of Pakistan. The activities of detoxifying enzymes i.e., β-esterases, GSTs and monooxygenases were also estimated. Our results revealed 100 percent mortality in insecticide treated groups after 24 hours of exposure. There was no difference in the activities of insecticide detoxifying enzymes between control and treated groups. We concluded from our work that malathion is still effective against C. quinquefasciatuns in the area for control of mosquito population.

The objective of this study was to evaluate the physiological response of the cabbage head caterpillar Crocidolomia pavonana treated with an active fraction of Calophyllum soulattri bark extract. Extraction of the test plant materials were performed with maceration method using methanol, continued by counter-current distribution separation in ethylacetate and water. Methanol fractionation of C. soulattri was performed by vaccuum liquid chromatography and the bioassays were conducted by a leaf-feeding method. The results showed that the dichloromethane fraction of C. soulattri had strong insecticidal activity against C. pavonana larvae, with LC 50 of 0.05%. Sublethal treatments with the active fraction at LC 15 , LC 50 , and LC 85 reduced the relative growth rate of the fourth instars by 48.9-94.1%. The treatments with the fraction at LC 15 and LC 50 to the fourth instars reduced the activity of invertase and protease enzyme by 20.7-24.1 and 14.4-25.14%, respectively, but increased the activity of trehalase by 26.7-120% as compared with controls.

This paper proposes a novel method to detect any outlier in a time-series data by analyzing the data in frequency domain and the chunk mean taken in the time domain. The combination of these two techniques increases the accuracy in detecting an outlier. The idea behind this approach is that whenever there is an outlier in the time series data, information would shift in higher frequency with greater magnitude. Thus, detecting this information will help to detect an outlier. This technique is not specific to a particular source generating time-series data. Instead, it is a generic approach to detect an outlier in any type of time-series data.

Application of the bioactive extract of the hutun seed (Barringtonia asiatica L.kurz) as an antifeedant in the Plutella xylostella Lcaterpillar pest on sawi plants (Brassica juncea L.) was carried out. This study aims to determine the antifeedant activity of the hutun seed extract in controlling the caterpillar sawi Plutella xylostella. The method used in this research is a completely randomized design (CRD) method with 4 treatments and 3 repetitions. The concentrations used are 50 ppm, 100 ppm, 500 ppm and 1000 ppm as well as positive control and negative control. The results of the study were tested with one-way ANOVA and continued with the Least Significant Difference test (LSD). Results of the study show that there are differences antifeedant activity caterpillar pests Plutella xylostella on sawi plants that significant at different levels of concentration. The study was conducted in several stages starting from the stage of extraction of the hutun seed, phytochemical test, preparation of caterpillar test, antifeedant activity testing. As the treatment is the level of methanol concentration of 50 ppm, 100 ppm, 500 ppm and 1000 ppm. The parameter observed was the percentage of Feeding Reduction (FR). The test results showed antifeedant activity against caterpillars of Plutella xylostella supreme contained at a concentration of 500 ppm and 1000 ppm because it can hinder eating caterpillars test amounted to 27.80 % and 40.93 %.

After more than 70 years of chemical pesticide use, modern agriculture is increasingly using biological control products. Resistances to conventional insecticides are wide spread, while those to bio-insecticides have raised less attention, and resistance management is frequently neglected. However, a good knowledge of the limitations of a new technique often provides greater sustainability. In this review, we compile cases of resistance to widely used bio-insecticides and describe the associated resistance mechanisms. This overview shows that all widely used bio-insecticides ultimately select resistant individuals. For example, at least 27 species of insects have been described as resistant to Bacillus thuringiensis toxins. The resistance mechanisms are at least as diverse as those that are involved in resistance to chemical insecticides, some of them being common to bio-insecticides and chemical insecticides. This analysis highlights the specific properties of bio-insecticides that the scientific community should use to provide a better sustainability of these products.

Since the discovery of vitamin E in 1922, its deficiency has been associated with various disorders, particularly atherosclerosis, ischemic heart disease, and the development of different types of cancer. A neurological syndrome associated with vitamin E deficiency resembling Friedreich ataxia has also been described. Whereas epidemiological studies have indicated the role of vitamin E in preventing the progression of atherosclerosis and cancer, intervention trials have produced contradictory results, indicating strong protection in some cases and no significant effect in others. Although it is commonly believed that phenolic compounds like vitamin E exert only a protective role against free radical damage, antioxidant molecules can exert other biological functions. For instance, the antioxidant activity of 17-␤-estradiol is not related to its role in determining secondary sexual characters, and the antioxidant capacity of all-trans-retinal is distinguished from its role in rhodopsin and vision. Thus, it is not unusual that ␣-tocopherol (the most active form of vitamin E) has properties independent of its antioxidant/radical scavenging ability. The Roman god Janus, shown in ancient coins as having two faces in one body, inspired the designation of 'Janus molecules' for these substances. The new biochemical face of vitamin E was first described in 1991, with an inhibitory effect on cell proliferation and protein kinase C activity. After a decade, this nonantioxidant role of vitamin E is well established, as confirmed by authoritative studies of signal transduction and gene regulation. More recently, a tocopherol binding protein with possible receptor function has been discovered. Despite such important developments in understanding the molecular mechanism and the targets of vitamin E, its new Janus face is not fully elucidated. Greater knowledge of the molecular events related to vitamin E will help in selecting the parameters for clinical intervention studies such as population type, dose response effects, and possible synergism with other compounds.-Ricciarelli, R.,

Objective: Glutathione transferases (GST) are multifunctional enzymes involved in detoxication, drug resistance, cell signaling and apoptosis. The inhibitory effects of novel benzazole derivatives were tested on human GST P1-1 to find new agents for overcoming drug resistance in cancer cells. Methods: GST P1-1 was heterogously expressed in E. coli strain XL-1 Blue and purified using S-hexylglutathione-Sepharose 6B affinity chromatography. The effect of 33 potential inhibitors on enzymatic activity was assayed spectrophotometrically with 1-chloro-2,4-dinitrobenzene (CDNB) as well as with the alternative substrate phenethyl isothiocyanate (PEITC). Results: Compound-18 (N-[2-(4-chloro-benzyl)-benzooxazol-5-yl]-4-nitro-benzenesulfonamide) was the most potent inhibitor found with an IC 50 value of approximately 10 µM with respect to CDNB and a somewhat less strong inhibitor (45 % inhibition at 40 µM) with PEITC as substrate. Compound-18 showed mixed inhibition with GSH and uncompetitive inhibition with CDNB with the K i values 6.3 ± 0.7 µM and 11.8 ± 3.4 µM, respectively. Conclusion: Compound-18 is a potent inhibitor of GST P1-1. It may serve as a lead for further chemical modifications for increased potency. Additional studies will elucidate the effects of the inhibitor on cancer cells.