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Trametes villosa Lignin Peroxidase (TvLiP): Genetic and Molecular Characterization

  • Carneiro, Rita Terezinha de Oliveira;Lopes, Maiza Alves;Silva, Marilia Lordelo Cardoso;Santos, Veronica da Silva;Souza, Volnei Brito de;Sousa, Aurizangela Oliveira de;Pirovani, Carlos Priminho;Koblitz, Maria Gabriela Bello;Benevides, Raquel Guimaraes;Goes-Neto, Aristoteles
    • Journal of Microbiology and Biotechnology
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    • 제27권1호
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    • pp.179-188
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    • 2017
  • White-rot basidiomycetes are the organisms that decompose lignin most efficiently, and Trametes villosa is a promising species for ligninolytic enzyme production. There are several publications on T. villosa applications for lignin degradation regarding the expression and secretion of laccase and manganese peroxidase (MnP) but no reports on the identification and characterization of lignin peroxidase (LiP), a relevant enzyme for the efficient breakdown of lignin. The object of this study was to identify and partially characterize, for the first time, gDNA, mRNA, and the corresponding lignin peroxidase (TvLiP) protein from T. villosa strain CCMB561 from the Brazilian semiarid region. The presence of ligninolytic enzymes produced by this strain grown in inducer media was qualitatively and quantitatively analyzed by spectrophotometry, qPCR, and dye fading using Remazol Brilliant Blue R. The spectrophotometric analysis showed that LiP activity was higher than that of MnP. The greatest LiP expression as measured by qPCR occurred on the $7^{th}$ day, and the ABSA medium (agar, sugarcane bagasse, and ammonium sulfate) was the best that favored LiP expression. The amplification of the TvLiP gene median region covering approximately 50% of the T. versicolor LPGIV gene (87% identity); the presence of Trp199, Leu115, Asp193, Trp199, and Ala203 in the translated amplicon of the T. villosa mRNA; and the close phylogenetic relationship between TvLiP and T. versicolor LiP all indicate that the target enzyme is a lignin peroxidase. Therefore, T. villosa CCMB561 has great potential for use as a LiP, MnP, and Lac producer for industrial applications.

Impact of Genetically Modified Enterobacter cloacae on Indigenous Endophytic Community of Citrus sinensis Seedlings

  • Fernando Dini;Mortatti, Marcelo-Jose;Souza, Andre-Oliveira de;Walter Maccheroni;Joao Lucio;Welington Luiz
    • Journal of Microbiology
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    • 제42권3호
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    • pp.169-173
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    • 2004
  • Enterobacter cloacae (strain PR2/7), a genetically modified endophyte(GME) in citrus plants, carrying different plasmids (pEC3.0/18, pCelE, pEglA and pGFP), was inoculated into Citrus sinensis seedlings under greenhouse conditions. The impact of this on the indigenous bacterial endophytic community was studied by analyses of 2 different morphologic groups. The germination rates of inoculated seeds were evaluated in greenhouse, and plasmid stability under in vitro conditions. Results demonstrated a great and diverse endophytic community inside plants, and specialization in tissue colonization by some bacterial groups, in different treatments. Shifts in seed germination rate were observed among treatments: in general, the PR2/7 harboring pEglA bacterial- clone significantly reduced seed germination, compared to the PR2/7 harboring pEC3.0/18 clone. This suggests that the presence of the pEglA plasmid changes bacteria-seed interactions. The endophytic community of citrus seedlings changed according to treatment. In seedlings treated with the PR2/7 with pEglA clone, the population of group II decreased significantly, within the context of the total endophytic community. These results indicate that the application of GMEs induces shifts in the endophytic bacterial community of citrus seedlings.

Prostate Cancer, High Cortisol Levels and Complex Hormonal Interaction

  • Fabre, Bibiana;Grosman, Halina;Gonzalez, Diego;Machulsky, Nahuel Fernandez;Repetto, Esteban M;Mesch, Viviana;Lopez, Miguel Angel;Mazza, Osvaldo;Berg, Gabriela
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권7호
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    • pp.3167-3171
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    • 2016
  • Prostate cancer (PCa) is one of the most common diseases in men. It is important to assess prognostic factors and whether high cortisol levels and complex hormonal interactions could be responsible for PCa development. We evaluated the relationship between cortisol, leptin and estrogens in 141 men, 71 with PCa and the remaining 70 constituting a low risk group (LRG). They were recruited for this study from a total of 2906 middle-aged men (ages 45-70 years) who completed an evaluation for prostatic diseases at the Urology Division, Hospital de Clínicas "$Jos{\acute{e}}$ de San $Mart{\acute{i}}n$", University of Buenos Aires, in May 2009. In this cross sectional study, cortisol, PSA, total-testosterone, free-testosterone, bioavailable testosterone, LH and estradiol were measured in serum. We observed increased cortisol levels in PCa patients as compared to LRG cases (p=0.004,). Leptin and estradiol levels were also higher in PCa patients (p=0.048; p<0.0001, respectively). Logistic regression analysis indicated that serum cortisol (OR: 1.110 (95% CI 1.016-1.213), p=0.022), estradiol (OR: 1.044 (95% CI 1.008-1.081), p=0.016) and leptin (OR: 1.248 (95% CI 1.048-1.487), p=0.013) explained 27% of the variance of dependent variables, even after adjusting for age, smoking, BMI and waist circumference. We found increased cortisol levels in PCa patients as compared to LRG, as well as an altered circulating hormonal profile.

Plant responses to nano and micro structured carbon allotropes: Water imbibition by maize seeds upon exposure to multiwalled carbon nanotubes and activated carbon

  • Dasgupta-Schubert, N.;Tiwari, D.K.;Francis, E. Reyes;Martinez Torres, P.;Villasenor Cendejas, L.M.;Lara Romero, J.;Villasenor Mora, C.
    • Advances in nano research
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    • 제5권3호
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    • pp.245-251
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    • 2017
  • Multiwalled carbon-nanotubes (MWCNT) and micro-structured carbon, such as biochar or activated carbon (AC), have been seen to significantly increase the growth indices of certain plant species such as maize (Zea mays L.). Seed imbibition is the stage where environmental factors that affect water transport across the seed coat barrier, make a large impact. This work explores the effect on water imbibition by maize seeds when the aqueous environment surrounding the seed is diluted by small concentrations (10 and 20 mg/l) of pristine MWCNT (p-MWCNT), carboxylate functionalized MWCNT (COO-MWCNT) and AC. The degree of sensitivity of the process to (i) large structural changes is seen by utilizing the nano (the MWCNT) and the micro (the AC) allotropic forms of carbon; (ii) to small changes in the purity and morphology of the p-MWCNT by utilizing 95% pure and 99% pure p-MWCNTs of slightly differing morphologies; and (iii) to MWCNT functionalization by using highly pure (97%) COO-MWCNT. Water imbibition was monitored over a 15 hour period by Near Infrared Thermography (NIRT) and also by seed weighing. Seed surface topography was seen by SEM imaging. Analysis of the NIRT images suggests rapid seed surface topological changes with the quantity of water imbibed. While further work is necessary to arrive at a conclusive answer, this work shows that the imbibition phase of the maize seed is sensitive to the presence of MWCNT even to small differences in the purity of the p-MWCNT and to small differences in the physicochemical properties of the medium caused by the hydrophilic COO-MWCNT.

Increased Expression of P2RY2, CD248 and EphB1 in Gastric Cancers from Chilean Patients

  • Aquea, Gisela;Bresky, Gustavo;Lancellotti, Domingo;Madariaga, Juan Andres;Zaffiri, Vittorio;Urzua, Ulises;Haberle, Sergio;Bernal, Giuliano
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권5호
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    • pp.1931-1936
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    • 2014
  • Background: Gastric cancer (GC) ranks as one of the major causes of mortality due to cancer worldwide. In Chile, it is currently the leading cause of cancer death. Identification of novel molecular markers that may help to improve disease diagnosis at early stages is imperative. Materials and Methods: Using whole-genome DNA microarrays we determined differential mRNA levels in fresh human GC samples compared to adjacent healthy mucosa from the same patients. Genes significantly overexpressed in GC were validated by RT-PCR in a group of 14 GC cases. Results: The genes CD248, NSD1, RAB17, ABCG8, Ephb1 and P2RY2 were detected as the top overexpressed in GC biopsies. P2RY2, Ephb1 and CD248 showed the best sensitivity for GC detection with values of 92.9%, 85.7% and 64.3% (p<0.05), respectively. Specificity was 85.7%, 71.4% and 71.4% (p<0.05), for each respectively.

Purification and Characterization of an Extracellular $\beta$-Glucosidase from Monascus purpureus

  • Daroit, Daniel J.;Simonetti, Aline;Hertz, Plinho F.;Brandelli, Adriano
    • Journal of Microbiology and Biotechnology
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    • 제18권5호
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    • pp.933-941
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    • 2008
  • An extracellular $\beta$-glucosidase produced by Monascus purpureus NRRL1992 in submerged cultivation was purified by acetone precipitation, gel filtration, and hydrophobic interaction chromatography, resulting in a purification factor of 92-fold. A $2^2$ central-composite design (CCD) was performed to find the best temperature and pH conditions for enzyme activity. Maximum activity was observed in a wide range of temperature and pH values, with optimal conditions set at $50^{\circ}C$ and pH 5.5. The $\beta$-glucosidase showed moderate thermostability, was inhibited by $HgCl_2$, $K_2Cr_O_4$, and $K_2Cr_2O_7$, whereas other reagents including $\beta$-mercaptoethanol, SDS, and EDTA showed no effect. Activity was slightly stimulated by low concentrations of ethanol and methanol. Hydrolysis of p-nitrophenyl-$\beta$-D-glucopyranoside (pNPG), cellobiose, salicin, n-octyl-$\beta$-D-glucopyranoside, and maltose indicates that the $\beta$-glucosidase has broad substrate specificity. Apparently, glucosyl residues were removed from the nonreducing end of p-nitrophenyl-$\beta$-D-cellobiose. $\beta$-Glucosidase affinity and hydrolytic efficiency were higher for pNPG, followed by maltose and cellobiose. Glucose and cellobiose competitively inhibited pNPG hydrolysis.

Elicitation of Penicillin Biosynthesis by Alginate in Penicillium chrysogenum, Exerted on pcbAB, pcbC, and penDE Genes at the Transcriptional Level

  • Liu, Gang;Casqueiro, Javier;Gutierrez, Santiago;Kosalkova, Katarina;Castillo, Nancy-Isabel;Martin, Juan-F.
    • Journal of Microbiology and Biotechnology
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    • 제11권5호
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    • pp.812-818
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    • 2001
  • Alginate and alginate-derived oligomannuronate enhanced penicillin production in shake flask and fermentor cultures of Penicillium chrysogenum Wis 54-1255 (containing a single copy of the penicillin gene cluster) and in the high producter strain P. chrysogenum AS-P-99 (containing multiple copies of the penicillin gene cluster). Alginate was not used as a single carbon source by P. chryogenum. The stimulatory effect on penicillin production was observed in a defined medium and, to a lower extent, in a complex production medium containing corn steep liquor. Alginate-supplemented cells showed higher transcript levels of the three penicillin biosynthetic genes, pcbAB, pcbC, and penDE, than cells grown in the absence of alginate. The promoters of the pcbAB, pcbC, and penDE genes were coupled to the reporter lacZ gene and introduced as monocopy constructions in P. chrysogenum Wis 54-1225 npe10 by targeted integration in the pyrG locus; the reporter ${\beta}$-galactosidase activity expressed from the three promoters was stimulated by alginate added to the culture medium of the transformants. These results indicate that the stimulation of penicillin production by alginate was derived from an increase in the transcriptional activity of the penicillin biosynthesis genes. The induction by alginate of the transcription of the three penicillin biosynthetic genes is good example of the coordinated induction of secondary metabolism genes by elicitors of plant (or microbial) origin.

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Taxonomic Reassessment of the Genus Padina (Dictyotales, Phaeophyta) from the Gulf of California

  • Riosmena-Rodriguez, Rafael;Paul-Chavez, Litzia;Hernandez-Carmona, Gustavo;Lopez-Vivas, Juan Manuel;Casas-Valdez, Margarita
    • ALGAE
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    • 제24권4호
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    • pp.213-229
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    • 2009
  • A monographic reassessment of Padina Adanson species from the Gulf of California was developed based on an exhaustive review of the character and characters states that have been used to delimit species in recent and historically valuable collections. Eight names (P. caulescens Thivy in Taylor, P. concrescens Thivy in Taylor, P. crispata Thivy in Taylor, P. durvillaei Bory Saint-Vincent, P. gymnospora [Kutzing] Sonder, P. mexicana Dawson, P. tetrastromatica Hauck and P. vickersiae Hoyt in Howe) were reported from the region. An analysis of 1,200 specimens led to the conclusion that only three species (P. durvillaei, P. concrescens and P. mexicana) can be recognized for the area based on four discriminating characters: presence of calcium carbonate in the cell wall; number of medullary cell layers in the basal area; number of medullary cell layers in the middle part and presence/absence of cortical cells. Comparative analysis of the species in the Gulf of California in relation to other recently evaluated species has shown a clear distinction among them suggesting Padina species were overestimated in our area.

Effect of mechanically deboned poultry meat content on technological properties and sensory characteristics of lamb and mutton sausages

  • Massingue, Armando Abel;Filho, Robledo de Almeida Torres;Fontes, Paulo Rogerio;Ramos, Alcineia de Lemos Souza;Fontes, Edimar Aparecida Filomeno;Perez, Juan Ramon Olalquiaga;Ramos, Eduardo Mendes
    • Asian-Australasian Journal of Animal Sciences
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    • 제31권4호
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    • pp.576-584
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    • 2018
  • Objective: This study aimed to develop a value-added product concerning technological and sensory characteristics changes of the use of mechanically deboned poultry meat (MDPM) as meat replacer in lamb and mutton emulsion-type sausages (mortadella). Methods: Sausages were produced with lamb and mutton and with different contents of MDPM. Six treatments, using lamb or mutton and 0%, 30%, and 60% of MDPM in relation to the meat batter, were produced and analyzed for pH, proximal composition, calcium and residual nitrite content, water activity, 2-thiobarbituric acid reactive substances (TBARS), instrumental color and texture profile. The sensory profile of the mortadella's was also evaluated by acceptance test and check-all-that-applies (CATA) analysis. Results: The MDPM addition increased (p<0.05) fat, residual nitrite and calcium content in the all sausage formulations, but mutton sausage had (p<0.05) higher fat and lower moisture content than lamb sausage. The pH, water activity, TBARS index and color was not affected by MDPM additions, while the mutton sausages were significantly redder (higher $a^*$, $C^*$, and lower $h^{\circ}$) and darker (lower $L^*$) than lamb sausages. Adding up to 60% of MDPM reduced (p<0.05) sausages hardness and chewiness. Overall, the meat replacement by MDPM increased the sausages acceptance, but the mutton sausage with 30% of MDPM replacer were the most preferred. Consumers related that pink color, glossy appearance, poultry meat-like taste, soft texture, juicy and greasy mouth feel to all sausages contain MDPM according to CATA analysis. Conclusion: Mutton from culled ewes can be utilized for mortadella production with 30% replacement of lean mutton and fat by MDPM.

P.W.R 형(型) 원자로(原子爐)의 증기발생기배수(蒸氣發生器排水)의 Ion 교환처리(交換處理) 연구(硏究) (Traitements sur $R\'{e}sines$ des Euax de Purge de $G\'{e}n\'{e}rateur$ de P. W. R. $Condition\'{e}es$ $\`{a}$ L' Ammoniaque)

  • 이상훈;박상훈
    • Journal of Radiation Protection and Research
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    • 제2권1호
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    • pp.24-30
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    • 1977
  • P.W.R. 형 원자로 증기발생기에서 발생되는 배수는 방사성 물질에 의하여 오염된다. 일반적으로 이 원자로의 증기발생기에 공급되는 급수는 'Phosphate'로 pH를 조절했으나 근래에는 'Ammonia' 조절방법이 선진제국에서 각광을 받고 있는 실정인 바 이는 약알카리로 pH를 조절하여 장치의 부식을 방지하기 위함이다. 본 실험에서는 이온 교환 수지탑을 이용하여 'Ammonia'로 조절된 배수를 처리하는 실험을 하였는 바, Acid Form의 Carboxylic, Cationic 몇 Anionic Ion Exchange Resinces Columns의 직열상태로 운전하였을 시 $^{137}Cs$$^{90}Sr$의 제염계수(Decontamination Factor)가 가장 높은 값을 나타낼 수 있으며 또한 그 처리능력이 가장 양호한 편으로 판명되었다.

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