90 research outputs found

    Short communication: Gender and heat stress effects on hypothalamic gene expression and feed intake in broilers

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    Our study aims to evaluate gender and heat stress effects on animal performance and on the expression of five hypothalamic genes related to feed consumption: neuropeptide Y (NPY), ghrelin (GHRL), pro-opiomelanocortin (POMC), AMP-activated protein kinase (AMPKα-1), and liver kinase B1 (LKB1). To assay these effects, 42-day-old male and female broilers were maintained in thermal comfort or were subjected to heat stress (HS, 38°C for 24 hours). All animals were fed with diets formulated to meet their nutritional requirements. Broilers subjected to HS showed lower weight gain (p=0.0065) and tended to have lower feed intake (p=0.0687) than broilers kept in comfortable conditions. We observed gender and heat stress interaction effects on NPY (p=0.0225), AMPKα-1 (p=0.0398), and POMC expression (p=0.0072). The highest NPY gene expression was observed in male broilers from the thermal comfort group. Male broilers exposed to HS showed the highest AMPKα-1 gene expression levels. Comparing POMC expression between males and females at the comfortable temperature, we observed that females showed higher POMC expression levels than male broilers. A gender effect was also observed on LKB1 and AMPKα-1 gene expression (p=0.0256 and p=0.0001, respectively); increased expression was observed in male broilers. Our results indicate that the expression of some hypothalamic genes related to food consumption may contribute to the observed differences in voluntary feed intake between animals of different gender exposed to different environmental conditions

    Analysis Of The Ergosterol Biosynthesis Pathway Cloning, Molecular Characterization And Phylogeny Of Lanosterol 14 α-demethylase (erg11) Gene Of Moniliophthora Perniciosa

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    The phytopathogenic fungus Moniliophthora perniciosa (Stahel) Aime & Philips-Mora, causal agent of witches’ broom disease of cocoa, causes countless damage to cocoa production in Brazil. Molecular studies have attempted to identify genes that play important roles in fungal survival and virulence. In this study, sequences deposited in the M. perniciosa Genome Sequencing Project database were analyzed to identify potential biological targets. For the first time, the ergosterol biosynthetic pathway in M. perniciosa was studied and the lanosterol 14α-demethylase gene (ERG11) that encodes the main enzyme of this pathway and is a target for fungicides was cloned, characterized molecularly and its phylogeny analyzed.ERG11 genomic DNA and cDNA were characterized and sequence analysis of the ERG11 protein identified highly conserved domains typical of this enzyme, such as SRS1, SRS4, EXXR and the heme-binding region (HBR). Comparison of the protein sequences and phylogenetic analysis revealed that the M. perniciosa enzyme was most closely related to that of Coprinopsis cinerea.374683693Aime, M.C., Phillips-Mora, W., The causal agents of witches’ broom and frost pod rot of cacao (chocolate, Theobroma cacao) from a new lineage of Marasmiaceae (2005) Mycologia, 97, pp. 1012-1022Albertini, C., Thebaud, G., Fournier, E., Leroux, P., Eburicol 14α-demethylase gene (CYP51) polymorphism and speciation in Botrytis cinerea (2002) Mycol Res, 106, pp. 1171-1178Altschul, S.F., Gish, W., Miller, W., Myersewand Lipman, D.J., Basic local alignment search tool (1990) J Mol Biol, 215, pp. 403-410Bak, S., Kahn, R.A., Oisen, C.E., Halkier, B.A., Cloning and expression in Escherichia coli of the obtusifoliol 14α-demethylase of Sorghum bicolor (L.) Moench, a cytochrome P450 orthologous to the sterol 14α demethylases (CYP51) from fungi and mammals (1997) Plant J, 11, pp. 191-201Barrett-Bee, K., Dixon, G., Ergosterol biosynthesis inhibition: A target for antifungal agents (1995) Acta Biochim Pol, 42, pp. 465-480Bellamine, A., Mangla, A.T., Nes, W.D., Waterman, M.R., Characterization and catalytic properties of the sterol 14α-demethylase from Mycobacterium tuberculosis (1999) Proc Natl Acad Sci USA, 96, pp. 8937-8942Butler, G., Rasmussen, M.D., Lin, M.F., Santos, M.A., Sakthikumar, S., Munro, C.A., Rheinbay, E., Reedy, J.L., Evolution of pathogenicity and sexual reproduction in eight Candida genomes (2009) Nature, 459, pp. 657-662Carrillo-Muñoz, A.J., Giusiano, G., Ezkurra, P.A., Quindós, G., Antifungal agents: Mode of action in yeast cells (2006) Rev Esp Quim, 19, pp. 130-139Ceita, G.O., Macedo, J.N., Santos, T.B., Alemanno, L., Gesteira, A.S., Micheli, F., Mariano, A.C., Meinhardt, L.W., Involvement of calcium oxalate degradation during programmed cell death in Theobroma cacao tissues triggered by the hemibiotrophic fungus Moniliophthora perniciosa (2007) Plant Sci, 173, pp. 106-117D’souza, C.A., Kronstad, J.W., Taylor, G., Warren, R., Yuen, M., Hu, G., Jung, W.H., Tangen, K., Genome variation in Cryptococcus gattii, an emerging pathogen of immunocompetent hosts (2011) MBio, 2, pp. e00342-e00410Délye, C., Laigret, F., Corio-Costet, M.F., Cloning and sequence analysis of the eburicol 14α-demethylase gene of the obligate biotrophic grape powdery mildew fungus (1997) Gene, 195, pp. 29-33Dujon, B., Sherman, D., Fischer, G., Durrens, P., Casaregola, S., Lafontaine, I., De Montigny, J., Talla, E., Genome evolution in yeasts (2004) Nature, 430, pp. 35-44Evans, H.C., Cacao diseases - The trilogy revisited (2007) Phytopathology, 97, pp. 1640-1643Felsenstein, J., Confidence limits on phylogenies: An approach using the bootstrap (1985) Evolution, 39, pp. 783-791Formighieri, E.F., Tiburcio, R.A., Armas, E.D., Medrano, F.J., Shimo, H., Carels, N., GóEs Neto, A., Sardinha-Pinto, N., The mitochondrial genome of the phytopathogenic basidiomycete Moniliophthora perniciosa is 109 kb in size and contains a stable integrated plasmid (2008) Mycol Res, 112, pp. 1136-1152Gasteiger, E., Hoogland, C., Gattiker, A., Duvaud, S., Wilkins, M.R., Appel, R.D., Bairoch, A., Protein identification and analysis tools on the ExPASy Server (2005) The Proteomics and Protocols Handbook, pp. 571-607. , In: Walker JM, Humana Press, TotowaGoffeau, A., Barrell, B.G., Bussey, H., Davis, R.W., Dujon, B., Feldmann, H., Galibert, F., Johnston, M., Life with 6000 genes (1996) Science, 265, pp. 2077-2082Griffith, G.W., Bravo-Velasquez, E., Wilson, F.J., Lewis, D.M., Hedger, J.N., Autecology and evolution of the witches’ broom pathogen (Crinipellis perniciosa) of cocoa (1994) The Ecology of Plant Pathogens, pp. 245-265. , In: Blakeman JP and Williamson B, CAB International, WallingfordGriffith, G.W., Nicholson, J., Neinninger, A., Birch, R., Witches’ brooms and frosty pods: Two major pathogens of cacao (2003) New Zeal J Bot, 41, pp. 423-435Hall, T.A., BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT (1999) Nucleic Acids Res, 41, pp. 95-98Hof, H., Critical annotations to the use of azole antifungals for plant protection (2001) Antimicrob Agents Chemother, 45, pp. 2987-2990Jackson, C.J., Lamb, D.C., Marczylo, T.H., Parker, J.E., Manning, N.L., Kelly, D.E., Kelly, S.L., Conservation and cloning of CYP51: A sterol 14α-demethylase from Mycobacterium smegmatis (2003) Biochem Biophys Res Commun, 301, pp. 558-563James, T.Y., Kauff, F., Schoch, C.L., Matheny, P.B., Hofstetter, V., Cox, C.J., Celio, G., Miadlikowska, J., Reconstructing the early evolution of Fungi using a six-gene phylogeny (2006) Nature, 19, pp. 818-822Jones, T., Federspiel, N.A., Chibana, H., Dungan, J., Kalman, S., Magee, B.B., Newport, G., Magee, P.T., The diploid genome sequence of Candida albicans (2004) Proc Natl Acad Sci USA, 101, pp. 7329-7334Kairuz, P.B., Zuber, J.P., Jaunin, P., Buchman, T.G., Bille, J., Rossier, M., Rapid detection and identification of Candida albicans and Torulopsis (Candida) glabrata in clinical specimens by species-specific nested PCR amplification of a cytochrome P-450 lanosterol-α-demethylase (L1A1) gene fragment (1994) J Clin Microbiol, 32, pp. 1902-1907Kalb, V.F., Woods, C.W., Turi, T.G., Dey, C.R., Sutter, T.R., Loper, J.C., Primary structure of the P450 lanosterol demethylase gene from Saccharomyces cerevisiae (1987) DNA, 6, pp. 529-537Kall, L., Krogh, A., Sonnhammer, E.L., Advantages of combined transmembrane topology and signal peptide prediction- the Phobius web server (2007) Nucleic Acids Res 35:429-, p. 432Kim, D., Lim, Y.R., Ohk, S.O., Kim, B.J., Chun, Y.J., Functional expression and characterization of CYP51 from dandruffcausing Malassezia globosa (2011) FEMS Yeast Res, 11, pp. 80-87Lai, M.H., Kirsch, D.R., Nucleotide sequence of cytochrome P450 L1A1 (lanosterol 14α-demethylase) from Candida albicans (1989) Nucleic Acids Res, 17, p. 804Lamb, D.C., Kelly, D.E., Manning, N.M., Hollomon, D.W., Kelly, S.L., Expression, purification, reconstitution and inhibition of Ustilago maydis sterol 14α-demethylase (CYP 51P450) (1998) FEMS Microbiol Lett, 169, pp. 369-373Lee, C.H., Hsu, K.H., Wang, S.Y., Chang, T.T., Chu, F.H., Shaw, J.F., Cloning and characterization of the lanosterol 14α-demethylase gene from Antrodia cinnamomea (2010) J Agr Food Chem, 58, pp. 4800-4807Lees, N.D., Skaggs, B., Kirsch DR and BirdM(1995) Cloning of the late genes in the ergosterol biosynthetic pathway of Saccharomyces cerevisiae - A review Lipids, 30, pp. 221-226Lepesheva, G.I., Waterman, M.R., Sterol 14α-demethylase cytochrome P450 (CYP51), a P450 in all biological kingdoms (2007) Biochim Biophys Acta, 3, pp. 467-477Luo, C.X., Schnabel, G., The cytochrome P450 lanosterol 14α-demethylase gene is a demethylation inhibitor fungicide resistance determinant in Monilia fructicola field isolates from Georgia (2008) Appl Environ Microb, 74, pp. 359-366Martin, F., Aerts, A., Ahrén, D., Brun, A., Danchin, E.G., Duchaussoy, F., Gibon, J., Pereda, V., The genome of Laccaria bicolor provides insights into mycorrhizal symbiosis (2008) Nature, 452, pp. 88-92McQuilken, M.P., Rudgard, S.A., Sensitivity of Crinipellis periciosa to two triazole fungicides in vitro and their effect on development of the fungus in cocoa (1988) Plant Pathol, 37, pp. 499-506Meinhardt, L.W., Bellato, C.M., Rincones, J., Azevedo, R.A., Cascardo, J.C.M., Pereira, G.A.G., In vitro production of biotrophic- like cultures of Crinipellis perniciosa, the causal agent of Witches’ broom disease of Theobroma cacao (2006) Curr Microbiol, 52, pp. 191-196Mellado, E., Guerra, T.M.D., Estrela, M.C., Tudela, J.L.R., Identification of two different 14α-sterol demethylase related genes (cyp51A and cyp51B) in Aspergillus fumigatus and other Aspergillus species (2001) J Clin Microbiol 39:2431-, p. 2438Mondego, J.M.C., Carazzolle, M.F., Costa, G.G.L., Formighieri, E.F., Parizzi, L.P., Rincones, J., Cotomacci, C., Carrer, H., A genome survey of Moniliophthora perniciosa gives new insights into Witches’ broom disease of cacao (2008) BMC Genomics, 9, pp. 1-25Morales, I.J., Vohra, P.K., Puri, V., Kottom, T.J., Limper, A.H., Thomas, C.F., Characterization of a lanosterol 14α demethylase from Pneumocystis carinii (2003) Am J Resp Cell Mol, 29, pp. 232-238Mota, S.G.R., Barros, T.F., Castilho, M.S., In vitro screening and chemometrics analysis on a series of azole derivativeswith fungicide activity against Moniliophthora perniciosa (2010) J Braz Chem Soc, 21, pp. 510-519Nierman, W.C., Pain, A., Anderson, M.J., Wortman, J.R., Kim, H.S., Arroyo, J., Berriman, M., Bermejo, C., Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus (2005) Nature, 438, pp. 1151-1156Page, R.D.M., TREEVIEW: An application to display phylogenetic trees on personal computers (1996) Comput Appl Biosci, 12, pp. 357-358Park, H.G., Lee, I.S., Chun, Y.J., Yun, C.H., Johnston, J.B., Montellano, P.R.O., Kim, D., Heterologous expression and characterization of the sterol 14α-demethylase CYP51F from Candida albicans (2011) Arch Biochem Biophys, 509, pp. 9-15Pereira, J.L., Ram, A., Figueiredo, J.M., Almeida, L.C.C., Primeira ocorrência de vassoura-de-bruxa na principal região produtora de cacau do Brasil (1989) Agrotrópica, 1, pp. 79-81Petersen, T.N., Brunak, S., Heijne, G., Nielsen, H., SignalIP 4.0: Discriminating signal peptides from transmembrane regions (2011) Nat Methods, 10, pp. 785-786Pietila, M.P., Vohra, P.K., Sanyat, B., Wengenack, N.L., Raghavakaimal, S., Thomas, C.F., Cloning and characterization of CYP51 from Mycobacterium avium (2006) Am J Resp Cell Mol, 35, pp. 236-240Pires, A.B.L., Gramacho, K.P., Silva, D.C., Góes-Neto, A., Silva, M.M., Muniz-Sobrinho, J.S., Porto, R.F., Cascardo, J.C.M., Early development of Moniliophthora perniciosa basidiomata and developmentally regulated genes (2009) BMC Microbiol, 9, p. e158Purdy, L.H., Schimidt, R.A., Status of cacao witches’ broom: Biology, epidemiology, and management (1996) Annu Rev Phytopathol, 34, pp. 573-594Raeder, U., Broda, P., Rapid preparation of DNA from filamentous fungi (1985) Lett Appl Microbiol, 1, pp. 17-20Revankar, S.G., Fu, J., Rinaldi, M.G., Kelly, S.L., Kelly, D.E., Lamb, D.C., Keller, S.M., Wickes, B.L., Cloning and characterization of the lanosterol 14α-demethylase (ERG11) gene in Cryptococcus neoformans (2004) Biochem Biophys Res Commun, 324, pp. 719-728Rincones, J., Scarpari, L.M., Carazzolle, M.F., Mondego, J.M.C., Formighieri, E.F., Barau, J.G., Costa, G.G.L., Vilas-Boas, L.A., Differential gene expression between the biotrophic-like and saprotrophic mycelia of the witches’ broom pathogen Moniliophthora perniciosa (2008) Mol Plant Microbe Int, 21, pp. 891-908Rio, M.C.S., Oliveira, B.V., Tomazella, D.P.T., Silva, J.A.F., Pereira, G.A.G., Production of calcium oxalate crystals by the basidiomycete Moniliophthora perniciosa, the causal agent of witches’ broom disease of cacao (2008) Curr Microbiol, 56, pp. 363-370Rozman, D., Stromstedt, M., Tsui, L.C., Scherer, S.W., Waterman, M.R., Structure and mapping of the human lanosterol 14α-demethylase gene (CYP51) encoding the cytochrome P450 involved in cholesterol biosynthesis: Comparison of exon/intron organization with other mammalian and fungal CYP genes (1996) Genomics, 38, pp. 371-381Sheng, C., Miao, Z., Ji, H., Yao, J., Wang, W., Che, X., Dong, G., Zhang, W., Three-dimensional model of lanosterol 14α-demethylase from Cryptococcus neoformans: Active- site characterization and insights into azole binding (2009) Antimicrob Agents Chemother, 53, pp. 3487-3495Sigrist, C.J.A., Cerutti, L., Castro, E., Langendijk-Genevaux, P.S., Bulliard, V., Bairoch, A., Hulo, N., PROSITE, a protein domain database for functional characterization and annotation (2009) Nucleic Acids Res, 38, pp. 161-166Stajich, J.E., Wilke, S.K., Ahrén, D., Au, C.H., Birren, B.W., Borodovsky, M., Burns, C., Cheng, C.K., Insights into evolution of multicellular fungi from the assembled chromosomes of the mushroom Coprinopsis cinerea (Coprinus cinereus) (2010) Proc Natl Acad Sci USA, 107, pp. 11889-11894Stanke, M., Keller, O., Gunduz, I., Hayes, A., Waack, S., Morgenstern, B., AUGUSTUS: Ab initio prediction of alternative transcripts (2006) Nucleic Acids Res, 34, pp. 435-439Swofford, D.L., PAUP - Phylogenetic Analysis Using Parsimony (and other methods). Version 4.0b10 (2002) Sinauer Associates, , Sunderland, MATer-Hovhannisyan, V., Lomsadze, L., Chernoff, Y.O., Borodovsky, M., Gene prediction in novel fungal genomes using an ab initio algorithm with unsupervised training (2008) Genome Res, 18, pp. 1979-1990Thompson, J.D., Higgins, D.G., Gibson, T.J., CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weigh matrix choice (1994) Nucleic Acids Res, 22, pp. 4673-4680Veen, M., Lang, C., Interactions of the ergosterol biosynthetic pathway with other lipid pathways (2005) Biochem Soc Trans, 33, pp. 1178-1181Warrilow, A.G.S., Melo, N., Martel, C.M., Parker, J.E., Nes, W.D., Kelly, S.L., Kelly, D.E., Expression, purification and characterization of Aspergillus fumigatus sterol 14α demethylase (CYP51) isoenzymes A and B (2010) Antimicrob Agents Chemother, 54, pp. 4225-4234Waterman, M.R., Lepesheva, G.I., Sterol 14 _-demethylase, an abundant and essential mixed-function oxidase (2005) Biochem Biophys Res Commun, 338, pp. 418-422Wood, H.M., Dickinson, M.J., Lucas, J.A., Dyer, P.S., Cloning of the CYP51 gene from the eyespot pathogen Tapesia yallundae indicates that resistance to the DMI fungicide prochloraz is not related to sequence changes in the gene encoding the target site enzyme (2001) FEMS Microbiol Lett, 196, pp. 183-187Zhao, L., Liu, D., Zhang, Q., Zhang, S., Wan J and XiaoW(2007) Expression and homology modeling of sterol 14α-demethylase from Penicillium digitatium FEMS Microbiol Lett, 277, pp. 37-4

    Purification, characterization and structural determination of UDP-N-acetylglucosamine pyrophosphorylase produced by Moniliophthora perniciosa

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    The enzyme UDP-N-acetylglucosamine pyrophosphorylase (PyroMp) from Moniliophthora perniciosa (CCMB 0257), a pathogenic fungal strain and the causative agent of the witches' broom disease in Theobroma cacao, was partially purified by precipitation with ammonium sulfate and gel filtration on Sephacryl S-200. The buffer for enzyme extraction was sodium phosphate, 0.050 mol L-1, pH 7.0, containing 1.0 mol L-1 NaCl. Response surface methodology (RSM) was used to determine the optimum pH and temperature conditions. Four different isoenzymes (PyroMp I, PyroMp II, PyroMp III and PyroMp IV) were obtained with optimal pH ranging from 6.9-8.4 and optimum temperature ranging from 28 to 68 °C. The 3D structure of pyrophosphorylase of M. perniciosa was determined by comparative modeling. The model obtained showed a good quality, possessing 78.6% of amino acids in energetically allowed regions. The model was then submitted for DM simulation and showed a good geometric quality (91.1% Ramachandran plot). The active site of the enzyme was found to be extremely well conserved. This model will be useful for developing new inhibitors against witches' broom disease22610151023CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFINANCIADORA DE ESTUDOS E PROJETOS - FINEPFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DA BAHIA - FAPESBFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informaçãoA enzima UDP-N-acetilglicosamina pirofosforilase de Moniliophthora perniciosa (CCMB 0257), o fungo patogênico causador da doença vassoura-de-bruxa do Theobroma cacao, foi parcialmente purificada por precipitação com sulfato de amônio e cromatografia de gel filtração em Sephacryl S-200. O tampão de extração da enzima foi o fosfato de sódio, 0,050 mol L-1, pH 7,0, contendo 1,0 mol L-1 de NaCl. A metodologia de superfície de resposta (MSR) foi usada para a obtenção do pH e temperatura ótima. Os resultados mostraram quatro diferentes isoenzimas (PyroMp I, PyroMp II, PyroMp III e PyroMp IV) que apresentaram pH ótimo na faixa de 6,9-8,4 e temperatura ótima variando entre 28 a 68 °C. A estrutura 3D de pirofosforilase de M. perniciosa foi obtida por modelagem comparativa. O modelo obtido mostrou uma boa qualidade, possuindo 78,6% de aminoácidos nas regiões energeticamente favoráveis. O modelo foi então submetido a simulações de dinâmica molecular (DM). O modelo apresentou uma boa qualidade geométrica após as simulações de DM (91,1% -gráfico de Ramachandran). A procura pelo sítio ativo da enzima mostrou que este é mantido extremamente conservado. Este modelo pode ser útil para desenvolvimento de inibidores contra a doença vassoura de brux

    The conceptual approach to the use of postbiotics based on bacterial membrane nanovesicles for prophylaxis of astronauts' health disorders

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    The functional fermented foods containing live microorganisms and their components are necessary for the normal functioning of the human body as normal gut microbiota needs fuel from external microbial organisms and their nanostructures — membrane vesicles (MVs), excreting outside. The сoncept that MVs may contribute to astronauts’ health probably to the same extent as their parental microbial cells do and be a temporary substitute for living microbial cells until we know more about the behavior of microbes in the space environment. The advantage of MVs is that they are not alive and cannot be changed under unfavorable conditions as microbial organisms may be. As the model, we selected MVs of a robust to environmental factors kombucha multimicrobial culture (KMC), known for its health-promoting characteristics for humans. We exposed KMC on the International Space Station in a hybrid space/Mars-like environment for an initial proof-of-concept stage. In the exposure study, KMC has survived a long-term period in harsh conditions, and the MVs generated by post-flight kombucha community members did not acquire toxicity, despite the changed membrane composition in the environment imitated conditions on the Mars surface. This observation, together with our KMC metagenomic and comparative genomic analyses of the dominant KMC bacterium Komagataeibacter oboediens, showed that the ground reference sample and spaceexposed ones were similar in topology and maintained their stability. In the next stage, we assessed the fitness, safety, and biodistribution of MVs of post-flight K. oboediens and showed that they were altered, but the modifications in membrane structure did not result in toxicity acquisition. Our proof-of-concept strategy is discussed in this review in line with the literature.The National Academy of Sciences of Ukraine Space Research Programme National Academy of Sciences of Ukraine Space Research Program.https://spj.science.org/journal/spaceam2023BiochemistryGeneticsMicrobiology and Plant Patholog
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