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Novel Protease from Aspergillus tamarii URM4634: Production and Characterization Using Inexpensive Agroindustrial Substrates by Solid-State Fermentation

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Advances in Enzyme Research 2016年第4期4卷 125-143页

作者： Osmar Soares da Silva Rodrigo Lira de Oliveira Cristina Maria souza-Motta Ana Lúcia Figueiredo Porto Tatiana souza Porto Northeast Biotechnology Network/RENORBIO Federal Rural University of Pernambuco (UFRPE) Recife Brazil Academic Unity of Garanhuns/UAG Federal Rural University of Pernambuco (UFRPE) Garanhuns Brazil Department Mycology Federal University of Pernambuco (UFPE) Recife Brazil Department of Morphology Physiology Animal/DMFA Federal Rural University of Pernambuco (UFRPE) Recife Brazil

This study reports the protease production from Aspergillus tamarii using agroindustrial residues as substrate for solid-state fermentation (SSF) and biochemical characterization. The highest protease production was obtained using wheat bran as substrate at 72 h fermentation with maximum proteolytic activity of 401.42 U/mL, collagenase of 243.0 U/mL and keratinase of 19.1 U/mL. The protease exhibited KM = 18.7 mg/mL and Vmax = 28.5 mg/mL/min. The optimal ph was 8.0 and stable in a wide ph range (5.0 - 11.0) during 24 h. The optimum temperature was 40°C. The proteolytic activity was inhibited by Cu2+ (33.98%) and hg2+ (22.69%). The enzyme was also inhibited by PMSF (65.11%), indicating that is a Serine Protease. These properties suggest that alkaline protease from A. tamarii URM4634 is suitable for application in food industries and leather processing. Additionally, the present findings opened new vistas in the utilization of wheat bran and other effective agroindustrial wastes as substrates for SSF.

关键词： Protease Aspergillus tamarii Biochemical Characterization Solid-State Fermentation Agroindustrial Waste

Association between helicobacterpylori, Epstein-Barr virus, human papillomavirus and gastric adenocarcinomas

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World Journal of Gastroenterology 2018年第43期24卷 4928-4938页

作者： Carolina Rosal Teixeira de souza Marcelli Carolini Alves Almeida Andre Salim Khayat Emerson Lucena da Silva Paulo Cardoso Soares Luiz Claudio Chaves Rommel Mario Rodriguez Burbano Laboratory of human Cytogenetics Institute of Biological Sciences Federal University of Para Belem Para 66075-110 Brazil Oncology Research Center Federal University of Para Joao deBarros Barreto University Hospital Belem Para 66073-000Brazil Ophir Loyola hospital Belem Para 66060-281 Brazil

AIM To correlate helicobacter pylori(h. pylori), Epstein-Barr virus(EBV) and human papillomavirus(hPV) with gastric cancer(GC) cases in Pará State, *** Tissue samples were obtained from 302 gastric adenocarcinomas. A rapid urease test was used to detect the presence of h. pylori, and the presence of the cagA gene in the hP-positive samples was confirmed by PCR. An RNA in situ hybridization test designed to complement Eber1 RNA was used to detect the presence of EBV in the samples, and the L1 region of hPV was detected using nested PCR. Positive hPV samples were genotyped and analyzed for E6 and E7 viral gene expression. Infections were also correlated with the clinical and pathological characteristics of the *** The majority of the 302 samples analyzed were obtained from men(65%) aged 55 years or older(67%) and were classified as the intestinal subtype(55%). All three pathogens were found in the samples analyzed in the present study(h. pylori : 87%, EBV: 20%, hPV: 3%). Overall, 78% of the h. pylori-positive(h. pylori+) samples were cag A +(h. pylori-cag A+), and there was an association between the cytotoxic product of this gene and EBV. Coinfections of h. pylori-cagA+ and EBV were correlated with the most advanced tumor stages. Although only 20% of the tumors were positive for EBV, infection with this virus was associated with distant metastasis. Only the hPV 16 and 18 strains were found in the samples, although no expression of the E6 and E7 oncoproteins was detected. The fundus of the stomach was the region least affected by the *** hPV was not involved in gastric tumorigenesis. Prophylactic and therapeutic measures against h. pylori and EBV may prevent the development of GC, especially the more aggressive forms.

关键词：乳头状瘤病毒协会腺癌胃区域检测测试设计 hPV EBV

Cryopreservation for delayed circulating tumor cell isolation is a valid strategy for prognostic association of circulating tumor cells in gastroesophageal cancer

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World Journal of Gastroenterology 2018年第7期24卷 810-818页

作者： Daniel Brungs David Lynch Alison WS Luk Elahe Minaei Marie Ranson Morteza Aghmesheh Kara L Vine Martin Carolan Mouhannad Jaber Paul de souza Therese M Becker Illawarra health and Medical Research Institute University of WollongongWollongong 2500Australia School of Biological Sciences University of WollongongWollongong 2500Australia Illawarra Cancer Centre Wollongong HospitalWollongong 2500Australia CONCERT-Translational Cancer Research Centre New South Wales 2000Australia Centre for Circulating Tumor Cell Diagnostics and Research Ingham Institute for Applied Medical ResearchLiverpool HospitalSydney 2170Australia School of Medicine University of Western SydneySydney 2170Australia South Western Medical School University of New South WalesSydney 2170Australia

AIM To demonstrate the feasibility of cryopreservation of peripheral blood mononuclear cells(PBMCs) for prognostic circulating tumor cell(CTC) detection in gastroesophageal *** Using 7.5 m L blood samples collected in EDTA tubes from patients with gastroesopheagal adenocarcinoma, CTCs were isolated by epithelial cell adhesion molecule based immunomagnetic capture using the Iso Flux platform. Paired specimens taken during the same blood draw(n = 15) were used to compare number of CTCs isolated from fresh and cryopreserved PBMCs. Blood samples were processed within 24 h to recover the PBMC fraction, with PBMCs used for fresh analysis immediately processed for CTC isolation. Cryopreservation of PBMCs lasted from 2 wk to 25.2 mo(median 14.6 mo). CTCs isolated from pre-treatment cryopreserved PBMCs(n = 43) were examined for associations with clinicopathological variables and survival *** While there was a significant trend to a decrease in CTC numbers associated with cryopreserved specimens(mean number of CTCs 34.4 vs 51.5, P = 0.04), this was predominately in samples with a total CTC count of > 50, with low CTC count samples less affected(P = 0.06). There was no significant association between the duration of cryopreservation and number of CTCs. In cryopreserved PBMCs from patient samples prior to treatment, a high CTC count(> 17) was associated with poorer overall survival(OS)(n = 43, hR = 4.4, 95%CI: 1.7-11.7, P = 0.0013). In multivariate analysis, after controlling for sex, age, stage, ECOG performance status, and primary tumor location, a high CTC count remained significantly associated with a poorer OS(hR = 3.7, 95%CI: 1.2-12.4, P = 0.03). CONCLUSION PBMC cryopreservation for delayed CTC isolation is a valid strategy to assist with sample collection, transporting and processing.

关键词： Cryopreservation Circulating tumor cells Liquid biopsy Gastroesophageal cancer Gastric cancer

Epidemiological Characteristics, Resistance Patterns and Spread of Gram-Negative Bacteria Related to Colonization of Patients in Intensive Care Units

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Advances in Infectious Diseases 2015年第1期5卷 14-20页

作者： Quésia souza Damaceno Jacques Nicoli Adriana Cristina Oliveira Departamento de Enfermagem Básica Escola de Enfermagem da Universidade Federal de Minas Gerais Belo Horizonte Brazil Departamento de Microbiologia Universidade Federal de Minas Gerais Belo Horizonte Brazil

Our aim was to determine the epidemiological characteristics, the resistance patterns and the spread of Gram negative bacteria related to colonization of patients in adult Intensive Care Units. Methods: A prospective cohort of patients colonized and/or infected with Gram negative bacteria was conducted at two adult ICUs from hospitals in Brazil (April 2012 to February 2013). Nasal, groin and perineum swabs were performed. Samples were incubated on MacConkey and cetrimide agar (48 h at 37℃) and identification tests (Vitek-BioMérieux), antibiogram (Bauer-Kirby method), Carba NP test, Polymerase Chain Reaction (PCR) and sequencing were performed. The patterns of resistant microorganisms were compared by rep-PCR (Diversilab). Results: There were 53 cases of colonization. In these cases, we identified imipenem-resistant Acinetobacter baumannii (51%), Pseudomonas aeruginosa (32%), Klebsiella pneumoniae ESBL (38%) or imipenem resistant (5.6%). The use of antimicrobials and medical devices were related to colonization (p The resistance patterns expressed by Klebsiella pneumoniae were ESBL (CTX-M, ShV e TEM) and KPC2. A verified profile of Acinetobacter baumannii was related to OXA-23 and OXA-253 (OXA-143 variant). The profiles ESBL and KPC2 expressed by Klebsiella pneumoniae were distributed between the both ICUs. The distribution of OXA-23 and OXA-253 was verified only in one ICU. The similarity of strains ranged from 80% to 95%, highlighting the horizontal transference of these microorganisms.

关键词： Intensive Care Unit Bacterial Drug-Resistance Gram-Negative Aerobic Bacteria Colonization

Resistance mechanism to cisplatin in NCI-h460 non-small cell lung cancer cell line:investigating apoptosis,autophagy,and cytogenetic damage

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Cancer Drug Resistance 2018年第1期1卷 72-81页

作者： Érica Ballestreri Daniel Simon Ana Paula de souza Camila Schultz Grott Débora Dreher Nabinger Rafael Rodrigues Dihl Ivana Grivicich Laboratory of Cancer Biology Graduate Program in Cellular and Molecular Biology Applied to HealthLutheran University of BrazilCanoasRS 92425-900Brazil Laboratory of human Molecular Genetics Graduate Program in Cellular and Molecular Biology Applied to HealthLutheran University of BrazilCanoasRS 92425-900Brazil Laboratory of Genetic Toxicity Graduate Program in Cellular and Molecular Biology Applied to HealthLutheran University of BrazilCanoasRS 92425-900Brazil

Aim:To investigate the effects of cisplatin on the human non-small cell lung carcinoma(NCI-h460)cell line regarding cytotoxicity,genotoxicity,and expression of genes associated with apoptosis(BIRC5)and autophagy(BECN1).Methods:Cell cultures were treated with cisplatin concentrations(0.16-33.3μmol/L)for 48 *** and acute and chronic cytotoxicities were assessed using the MTT,clonogenic,and cytokinesis-block micronucleus *** expression of BIRC5 and BECN1 was evaluated by reverse transcription-polymerase chain ***:Cisplatin IC50(0.33μmol/L)increased micronucleus frequency 2.50 *** was also cytotoxic in the 0.6-33.3μmol/L range,with reduced expression of the BIRC5 gene,suggesting induction of *** reducing the expression of the BIRC5 gene,33.3μmol/L cisplatin increased the expression of the BECN1 gene,suggesting that autophagy can be related to cisplatin ***:Cisplatin inhibited NCI-h460 growth,and cisplatin IC50 induced genotoxic *** higher cisplatin concentrations are used,the expression of genes associated with apoptosis and autophagy was *** results points to a further investigation of the role of autophagy in cisplatin resistance.

关键词： BECN1 BIRC5 cisplatin cytotoxicity lung cancer

萝卜硫烷对脑线粒体的影响:机理观点和未来发展方向

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2020年第4期21卷 263-279页

作者： Fernanda Rafaela JARDIM Fhelipe Jolner souza de ALMEIDA Matheus Dargesso LUCKAChAKI Marcos Roberto de OLIVEIRA Forensic Institute Forensic Toxicology DivisionPostmortem Toxicology SectorCEP 90160-093Porto AlegreRSBrazil Postgraduate Program in health Sciences(PPGCS) Federal University of Mato Grosso(UFMT)CEP 78060-900CuiabaMTBrazil Department of Chemistry Federal University of Mato Grosso(UFMT)CEP 78060-900CuiabaMTBrazil Postgraduate Program in Chemistry(PPGQ) Federal University of Mato Grosso(UFMT)CEP 78060-900CuiabaMTBrazil Department of Biochemistry Prof.“Tuiskon Dick” Federal University of Rio Grande do Sul(UFRGS)CEP 90035-000Porto AlegreRSBrazil

有机硫化物萝卜硫烷(SFN)是一种有效的细胞保护剂,在体外和体内实验模型中均可促进抗氧化、抗炎、抗糖化和抗菌作用。因为氧化磷酸化系统的存在,所以线粒体是三磷酸腺苷(ATP)的主要产生部位。同时,线粒体也是具核人体细胞中产生活性氧... 详细信息

有机硫化物萝卜硫烷(SFN)是一种有效的细胞保护剂,在体外和体内实验模型中均可促进抗氧化、抗炎、抗糖化和抗菌作用。因为氧化磷酸化系统的存在,所以线粒体是三磷酸腺苷(ATP)的主要产生部位。同时,线粒体也是具核人体细胞中产生活性氧的主要场所。线粒体损伤在多种人类疾病中起着重要作用,包括神经退行性病变和代谢异常。本文描述和讨论了SFN调节哺乳动物细胞中线粒体功能和动力学的作用和机理,以及其对肿瘤细胞中线粒体促凋亡途径的促进作用。SFN对线粒体的调节作用使得它在一定程度上被认为是一种细胞保护剂。本文还指出了几个需要通过进一步实验解决的问题,即未来的可能研究方向,这可能会对该领域的研究人员具有指导作用。

关键词：萝卜硫素脑线粒体抗氧化剂 Nrf2

Mangiferin,a natural xanthone,accelerates gastrointestinal transit in mice involving cholinergic mechanism

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World Journal of Gastroenterology 2012年第25期18卷 3207-3214页

作者： Talita Cavalcante Morais Synara Cavalcante Lopes Karine Maria Martins Bezerra Carvalho Bruno Rodrigues Arruda Francisco Thiago Correia de souza Maria Teresa Salles Trevisan Vietla Satyanarayana Rao Flávia Almeida Santos Department of Physiology and Pharmacology Brazilian SemiArid Institute of BiomedicineFaculty of MedicineFederal University of CearáFortaleza 60430-270Brazil Post-Graduate Programme in Medical Sciences Faculty of MedicineFederal University of CearáFortaleza 60430-140Brazil Department of Organic and Inorganic Chemistry Federal University of CearáFortaleza 60451-970Brazil

AIM:To investigate the effects of mangiferin on gastrointestinal transit(GIT) in normal and constipated mice,together with the possible ***:Intragastrically-administered charcoal mealwas used to measure GIT in overnight starved Swiss *** the first experiments,mangiferin(3 mg/kg,10 mg/kg,30 mg/kg,and 100 mg/kg,po) or tegaserod(1 mg/kg,ip) were administered 30 min before the charcoal meal to study their effects on normal *** the second series,mangiferin(30 mg/kg) was tested on delayed GIT induced by several different pharmacological agonists(morphine,clonidine,capsaicin) or antagonists(ondansetron,verapamil,and atropine) whereas in the third series,mangiferin(30 mg/kg,100 mg/kg and 300 mg/kg) or tegaserod(1 mg/kg) were tested on 6 h fecal pellets outputted by freely fed *** ratio of wet to dry weight was calculated and used as a marker of fecal water ***:Mangiferin administered orally significantly(P hydroxytryptamine4(5-hT4) agonist tegaserod(81%) when compared to vehicle-treated control(63%).Co-administered mangiferin(30 mg/kg) totally reversed the inhibitory effect of opioid agonist morphine,5-hT3-receptor antagonist ondansetron and transient receptor potential vanilloid-1 receptor agonist capsaicin on GIT,but only to a partial extent with the GIT-delay induced by 2-adrenoceptor agonist clonidine,and calcium antagonist ***,co-administered atropine completely blocked the stimulant effect of mangiferin on GIT,suggesting the involvement of muscarinic acetylcholine receptor *** mangiferin significantly enhanced the 6 h fecal output at higher doses(245.5 ± 10.43 mg vs 161.9 ± 10.82 mg and 227.1 ± 20.11 mg vs 161.9 ± 10.82 mg of vehicle-treated control,at 30 and 100 mg/kg,P he effect of tegaserod was more potent(297.4 ± 7.42 mg vs 161.9 ± 10.82 mg of vehicle-treated control,P < 0.05).Unlik

关键词：生理机制胃肠蠕动肾上腺素受体激动剂芒果小鼠氧杂蒽酮胆碱能受体拮抗剂

Conocarpus erectus L., a plant with a high content of structural sugars, ions and phenolic compounds, shows antioxidant and antimicrobial properties promoted by different organic fractions

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Asian Pacific Journal of Tropical Biomedicine 2018年第9期8卷 463-470页

作者： Dayane Kelly Dias do Nascimento Santos Weslley henrique de Oliveira Melo Anastassia Mariáh Nunes de Oliveira Lima Iranildo Joseda Cruz Filho Glaucia Manoella de souza Lima Tulio Diego da Silva Maiara Celine de Moura Marcia Silva do Nascimento Ana Maria Souto Maior Thiago henrique Napoleao Cristiane Moutinho Lagos de Melo Department of Biochemistry Biosciences Centre Federal University of Pernambuco Department of Antibiotics Biosciences Centre Federal University of Pernambuco Center of Strategic Technologies of the Northeast

Objective: To evaluate the structural and chemical composition of plant and the antioxidant and antimicrobial activities promoted by hexanic, ethanolic and ethyl acetate fractions obtained from leaves of Conocarpus ***: Organic fractions were characterized through UPLC-MS and *** potential was performed through DPPh and molybdenum phosphate *** and antifungal assays were performed in accordance with Clinical and Laboratory Standards Institute ***: The obtained biomass of Conocarpus erectus leaves showed the high presence of glucose(0.45 g/L), cellulose(28.69%), Na(55.126 μg/L) and K(31.163 μg/L).We identified seven compounds in the hexanic and ethyl acetate fractions, and eight compounds in ethanolic ***, phenolic compounds are prevalent in all organic fractions with values of(10.04 ± 0.24),(221.26 ± 1.84),(340.53 ± 0.84) mg/g GAE to hexanic, ethyl acetate and ethanolic fraction, *** results showed a high potential in ethyl acetate fraction(71.82 ± 6.87)% and(10.89 ± 0.05)% in DPPh and molybdenum phosphate techniques, *** ethanolic fraction showed moderate bacteriostatic and bactericidal activity against Staphylococcus aureus and presented a high fungistatic potential for all Candida species ***: Organic fractions obtained from leaves of Conocarpus erectus present antimicrobial and antioxidant properties, and these findings contribute to scientific information for the effectiveness on use of this plant in the development of a phytotherapic compound.

关键词： Combretaceae Phenolic compounds Antimicrobial activity Antioxidant

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Dictyosporiaceae ***.

Fungal Diversity 2016年第5期80卷 457-482页

作者： Saranyaphat Boonmee Melvina J.D’souza Zonglong Luo Umpava Pinruan Kazuaki Tanaka hongyan Su Darbhe J.Bhat Eric h.C.McKenzie E.B.Gareth Jones Joanne E.Taylor Alan J.L.Phillips Kazuyuki hirayama Prapassorn D.Eungwanichayapant Kevin D.hyde Center of Excellence in Fungal Research Mae Fah Luang UniversityChiang Rai 57100Thailand School of Science Mae Fah Luang UniversityChiang Rai 57100Thailand College of Agriculture and Biology Sciences Dali UniversityDaliYunnan 671003China National Center for Genetic Engineering and Biotechnology National Science and Technology Development AgencyThailand Science Park113 Phahonyothin RoadKlong 1Klong Luang Pathumthani 12120Thailand Formerly Department of BotanyGoa UniversityNo.128/1-JAzad Housing SocietyCurca Goa VelhaIndia Manaaki Whenua Landcare Research Private Bag 92170AucklandNew Zealand Faculty of Sciences Biosystems and Integrative Sciences Institute(BioISI)University of LisbonCampo Grande1749-016 LisbonPortugal Royal Botanic Garden Edinburgh 20A Inverleith RowEdinburgh EH35LRUK Department of Botany and Microbiology College of ScienceKing Saud UniversityP.O.Box:2455Riyadh 1145Saudi Arabia Faculty of Agriculture and Life Science Hirosaki University3 Bunkyo-choHirosakiAomori 036-8561Japan Apple Experiment Station Aomori Prefectural Agriculture and Forestry Research Center24 FukutamiBotandairaKuroishiAomori 036-0332Japan

Dictyosporiaceae *** introduced to accommodate a holomorphic group of Dothideomycetes that are saprobes on decaying wood and plant debris in terrestrial and freshwater *** sexual morph comprises Dictyosporium meiosporum,***,Gregarithecium curvisporum,Pseudocoleophoma calamagrostidis and *** sexual morphs are characterized by immersed to erumpent,superficial,globose to subglobose,dark brown to black ascomata,bitunicate asci,and septate,hyaline,sheathed *** asexual morphs are hyphomycetous with brown,multi-septate,cheirosporous conidia,produced from holoblastic conidiogenous cells,on micronematous *** analyses of LSU,SSU and TEF1 sequence data place the genera with cheirospores in a monophyletic clade in the order *** analyses based on ITS sequence data show the family to comprise 44 distinct lineages that correspond to ten genera and an undetermined *** new genera Dictyocheirospora,Dictyopalmispora,Jalapriya and Vikalpa,and eight new species Dictyocheirospora bannica,***,***,Dictyopalmispora palmae,Dictyosporium olivaceosporum,***,Jalapriya pulchra,and Vikalpa lignicola,are introduced in the family.

关键词： Cheirosporous conidia KeylPhylogeny Sexual states Sporodochial fungi