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Partial purification,immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus

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Advances in Enzyme Research 2013年第4期1卷 79-90页

作者： Ana Lucia Ferrarezi Daniele h.Pivetta gustavo Orlando Bonilla-Rodriguez Roberto da Silva Jose Manuel Guisan Eleni Gomes Benevides Costa Pessela Department of Biology Laboratory of Applied Biochemistry and MicrobiologyIBILCE-UNESPSao Jose do Rio Preto SPBrazil Department of Chemistry and Environmental Sciences IBILCE-UNESPSao Jose do Rio Preto SPBrazil Department of Enzyme Biocatalysis Institute of Catalysis and PetrochemistryCSICCampus UAMCantoblancoMadridSpain Department of Biotechnology and Food Microbiology Research Institute for Food ScienceCIAL-CSICCalleNicolás Cabrera 9Campus UAMMadridSpain

Lipases have important applications in biotechnological processes, motivating us to produce, purify, immobilize and perform a biochemical characterization of the lipase from Rhizomucor pusillus. The fungus was cultivated by solid state fermentation producing lipolytic activity of about 0.5 U/mL(4U/g). A partial purification by gel filtration chromatography in Se-phacryl S-100 allowed obtaining a yield of about 85% and a purification factor of 5.7. Our results revealed that the purified enzyme is very stable with some significant differences in its properties when compared to crude extract. The crude enzyme extract has an optimum ph and temperature of 7.5 ° C and 40 ° C, respectively. After purification, a shift of the optimum ph from 7 to 8 was observed, as well as a rise in optimumtemperature to 60 ° C and an increase in stability. The enzyme was immobilized on CNBr-Agarose and Octyl-Agarose supports, having the highest immobilization yield of 94% in the second resin. The major advantage of immobilization in hydrophobic media such as Octyl is in its hyper activation, which in this case was over 200%, a very interesting finding. Another advantage of this type of immobilization is the possibility of using the derivatives in biotechnological applications, such as in oil enriched with omega-3 as the results obtained in this study display the hydrolysis of 40% EPA and 7% DhA from sardine oil, promising results compared to the literature.

关键词： Enzyme Immobilization Solid State Fermentation Purification n-3 Polyunsatured Fatty Acids Rhizomucor pusillus

Medicinal properties of Angelica archangelica root extract: Cytotoxicity in breast cancer cells and its protective effects against in vivo tumor development

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Journal of Integrative Medicine 2019年第2期17卷 132-140页

作者： Carlos R.Oliveira Daniel G.Spindola Daniel M.Garcia Adolfo Erustes Alexandre Bechara Caroline Palmeira-dos-Santos Soraya S.Smaili gustavo J.S.Pereira André hinsberger Ezequiel P.Viriato Maria Cristina Marcucci Alexandra C.h.F.Sawaya Samantha L.Tomaz Elaine G.Rodrigues Claudia Bincoletto Departamento de Farmacologia Escola Paulista de Medicina (EPM) Universidade Federal de Sao Paulo (UNIFESP) Grupo de Fitocomplexos e Sinalizacao Celular Escola de Ciências da Saúde Universidade Anhembi Morumbi Faculdade de Ciências Farmacêuticas e Bioquímicas Oswaldo Cruz Laboratório de Produtos Naturais e Quimiometria Programa de pós-graduacao em Farmácia e Biotecnologia Universidade Anhanguera de Sao Paulo Faculdade de Ciências Farmacêuticas Universidade Estadual de Campinas (UNICAMP) Unidade de Oncologia Experimental EPM Universidade Federal de Sao Paulo (UNIFESP)

Objective: Although Angelica archangelica is a medicinal and aromatic plant with a long history of use for both medicinal and food purposes, there are no studies regarding the antineoplastic activity of its *** study aimed to evaluate the cytotoxicity and antitumor effects of the crude extract of A. archangelica root(CEAA) on breast ***: The cytotoxicity of CEAA against breast adenocarcinoma cells(4T1 and MCF-7) was evaluated by a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-h-tetrazolium bromide(MTT) assay. Morphological and biochemical changes were detected by hoechst 33342/propidium iodide(PI) and annexin V/PI staining. Cytosolic calcium mobilization was evaluated in cells staining with FURA-4 NW. Immunoblotting was used to determine the effect of CEAA on anti-and pro-apoptotic proteins(Bcl-2 and Bax, respectively).The 4 T1 cell-challenged mice were used for in vivo ***: Using ultra-high-performance liquid chromatography–mass spectrometry analysis, angelicin, a constituent of the roots and leaves of A. archangelica, was found to be the major constituent of the CEAA evaluated in this study(73 mg/mL). The CEAA was cytotoxic for both breast cancer cell lines studied but not for human fibroblasts. Treatment of 4 T1 cells with the CEAA increased Bax protein levels accompanied by decreased Bcl-2 expression, in the presence of cleaved caspase-3 and cytosolic calcium mobilization, suggesting mitochondrial involvement in breast cancer cell death induced by the CEAA in this cell line. No changes on the Bcl-2/Bax ratio were observed in CEAA-treated MCF7 cells. Gavage administration of the CEAA(500 mg/kg) to 4T1 cell-challenged mice significantly decreased tumor growth when compared with untreated ***: Altogether, our data show the antitumor potential of the CEAA against breast cancer cells in vitro and in vivo. Further research is necessary to better elucidate the pharmacological application of the CEAA in breast cancer therapy.

关键词： Angelica archangelica Umbelliferae Apoptosis Cell death Breast cancer Angelicin

Antitumor effects of electrochemical treatment

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Chinese Journal of Cancer Research 2013年第2期25卷 223-234页

作者： héctor Manuel Camué Ciria Maraelys Morales González Lisset Ortíz Zamora Luis Enrique Bergues Cabrales gustavo Victoriano Sierra González Luciana Oliveira de Oliveira Rodrigo Zanella Antonio Carlos Buzaid Orlando Parise Luciana Macedo Brito Cesar Augusto Antunes Teixeira Marina das Neves Gomes Gleyce Moreno Venicio Feo da Veiga Marcos Telló Carla holandino Research Department National Centre of Applied Electromagnetism Oriente University Pharmacy Department Oriente University Finlay Institute La lisa. La Habana Cuba Veterinary Teaching hospital Federal University of Rio Grande do Sul. Porto Alegre Sírio e Libanês hospital Faculty of Pharmacy Medicine Department Federal University of Rio de Janeiro Professor Paulo de Góes Institute of Microbiology Federal University of Rio de Janeiro Pontifical Catholic University of Rio Grande do Sul. Porto Alegre

Electrochemical treatment is an alternative modality for tumor treatment based on the application of a low intensity direct electric current to the tumor tissue through two or more platinum electrodes placed within the tumor zone or in the surrounding areas. This treatment is noted for its great effectiveness, minimal invasiveness and local effect. Several studies have been conducted worldwide to evaluate the anfitumoral effect of this therapy. In all these studies a variety of biochemical and physiological responses of tumors to the applied treatment have been obtained. By this reason, researchers have suggested various mechanisms to explain how direct electric current destroys tumor ceils. Although, it is generally accepted this treatment induces electrolysis, electroosmosis and electroporation in tumoral tissues. however, action mechanism of this alternative modality on the tumor tissue is not well understood. Although the principle of Electrochemical treatment is simple, a standardized method is not yet available. The mechanism by which Electrochemical treatment affects tumor growth and survival may represent more complex process. The present work analyzes the latest and most important research done on the electrochemical treatment of tumors. We conclude with our point of view about the destruction mechanism features of this alternative therapy. Also, we suggest some mechanisms and strategies from the thermodynamic point of view for this therapy. In the area of Electrochemical treatment of cancer this tool has been exploited very little and much work remains to be done. Electrochemical treatment constitutes a good therapeutic option for patients that have failed the conventional oncology methods.

关键词： Direct electric current electrochemical treatment antimmoral treatment tumor tumor electrolysis rumor entropy dissipative structure

Effects of Saccharomycesboulardiion fecal short-chain fatty acids and microflora in patients on long-term total enteral nutrition

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World Journal of Gastroenterology 2005年第39期11卷 6165-6169页

作者： Stéphane M Schneider Fernand Girard-Pipau Jér(o|^)me Filippi Xavier hébuterne Dominique Moyse gustavo Calle hinojosa Anne Pompei Patrick Rampal Department of hepato-Gastroenterology and Clinical Nutrition Archet University hospital NiceFrance Bacteriology Laboratory Archet University hospital NiceFrance Biostatistician Paris France Department of hepato-Gastroenterology Princess Grace hospital Monte-CarloMonaco

AIM: To assess the effects of Sb on fecal flora and shortchain fatty acids (SCFA) in patients on long-term ***: Ten patients (3 females, 7 males, 59±5.5 years),on TEN for a median of 13 mo (1-125), and 15 healthy volunteers (4 females, 11 males, 32±2.0 years) received Sb (0.5 g bid PO) for 6 d. Two stool samples were taken before, on the last 2 d and 9-10 d after treatment, for SCFA measurement and for culture and bacterial *** (mean±SE) were compared using sign tests and ***: Fecal butyrate levels were lower in patients(10.1±2.9 mmol/kg) than in controls (19.2±3.9, P= 0.02).Treatment with Sb increased total fecal SCFA levels in patients (150.2±27.2 vs 107.5±18.2 mmol/kg, P = 0.02)but not in controls (129.0±28.6 vs 113.0±15.2 mmol/kg,NS). At the end of treatment with Sb, patients had higher fecal butyrate (16.0±4.4 vs 10.1 [2.9] mmol/kg, P = 0.004).Total SCFAs remained high 9 d after treatment was discontinued. Before the treatment, the anaerobe to aerobe ratio was lower in patients compared to controls (2.4±2.3 vs69.8±1.8, P = 0.003). There were no significant changes in the fecal flora of TEN ***: Sb-induced increase of fecal SCFA concentrations (especially butyrate) may explain the preventive effects of this yeast on TEN-induced diarrhea.

关键词：酵母脂肪酸肠内营养腹泻

A positive feedback loop involving Gcm1 and Fzd5 directs chorionic branching morphogenesis in the placenta

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A positive feedback loop involving Gcm1 and Fzd5 directs cho...

Fisher's perceptions about a marine protected area over time

2012全国发育生物学大会

作者： haruo Nakano David G.Simmons Johan h.van Es hans Clevers gustavo Leone James C.Cross State key laboratory of Reproductive Biology Institute of ZoologyChinese Academy of Sciences

Chorioallantoic branching morphogenesis is a key milestone during placental development,creating the large surface area for nutrient and gas exchange and is therefore critical for the success of term *** Wnt pathway molecules have been shown to regulate placental ***,it remains largely unknown how Wnt-Frizzled (Fzd) signaling spatiotemporally interacts with other essential regulators,ensuring chorionic branching morphogenesis and angiogenesis during placental *** systemic and trophoblast-specific Fzd5 null and Gcm1 deficient mouse models,combining trophoblast stem cell lines and tetraploid aggregation assay,we demonstrate here that an amplifying signaling loop between Gcm1 and Fzd5 is essential for normal initiation of branching in the chorionic plate. While Gcm1 upregulates Fzd5 specifically at sites where branching initiates in the basal chorion,this elevated Fzd5 expression via nuclearβ-catenin signaling in turn maintains expression of ***,we show that Fzd5-mediated signaling induces the disassociation of cell junctions for branching initiation via downregulating ZO-1 expression in trophoblast cells at the base of the *** addition,Fzd5-mediated signaling is also important for upregulation of Vegf expression in chorion trophoblast ***,we demonstrate that Fzd5-Gcm1 signaling cascade is operative during human trophoblast differentiation. Conclusions:These data indicate that Gcm1 and Fzd5 function in an evolutionary conserved positive feedback loop that regulates trophoblast differentiation and sites of chorionic branching morphogenesis.

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Aquaculture and Fisheries 2020年第5期5卷 273-281页

作者： Cristina Pita Barbara horta e Costa gustavo Franco Rui Coelho Inês Sousa Emanuel J.Gonçalves Jorge M.S.Gonçalves Karim Erzini Department of Environment and Planning&Centre for Environmental and Marine Studies(CESAM) University of AveiroPortugal Center of Marine Sciences CCMARUniversity of AlgarveCampus de Gambelas8005-139FaroPortugal MARE-Marine and Environmental Sciences Centre ISPAInstituto UniversitárioRua Jardim Do Tabaco 341149-041LisboaPortugal Instituto Português Do Mar a da Atmosfera IP(IPMA)-Polo OlhãoAv.5 de Outubro8700-305OlhãoPortugal

The perceptions of fishers towards the Arrábida Marine Park,a marine protected area(MPA)in the west coast of Portugal,were studied through face-to-face interviews in two different moments of the MPA life ***'perceptions about the MPA and the impact it had on the fishing activity over time were identified just before the full implementation of the zoning and regulations of the management plan and 10 years *** study aimed to investigate fishers'knowledge,acceptance and perceptions about the MPA changed with time,if support for the MPA was linked to the impact of the MPA on the fishing activity,and if fishers'perceptions about the impact of the MPA on the fishing activity match with local landings *** show that although knowledge about the marine park significantly improved over time,fishers'acceptance did not.A decrease on fishers'support was not substantial but *** such as the disagreement with regulations reinforced concerns raised during the implementation of the marine park,particularly in relation to the top-down decision-making,which commonly confers minor participation,recognition and legitimacy to ***,fisheries benefits were still not perceived by local fishers,though they are central for fishers'***,the perceived negative impacts of the park seemed to be more related to social aspects and individual interests than to impacts on *** adequate management,enforcement and participation of local fishers is still possible and are advocated here as to contribute to the expected socioecological outcomes and respective support,leading to the future successful performance of the Arrábida Marine *** fishers’perceptions towards an MPA over time is central and should be included on periodical socioecological monitoring and inform an effective adaptive management.

关键词： Arrabida Marine Park(AMP) Local fisheries Perceptions Attitudes Fishers'support Decision-making Marine protected area Changes over time

hemoconcentration is a poor predictor of severity in acute pancreatitis

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World Journal of Gastroenterology 2005年第44期11卷 7018-7023页

作者： José M.Remes-Troche Andrés Duarte-Rojo gustavo Morales Guillermo Robles-Díaz Department of Gastroenterology

AIM: To determine whether the hematocrit (hct) at admission or at 24 h after admission was associated with severe acute pancreatitis (AP), organ failure (OF), and pancreatic ***: A total of 336 consecutive patients with a first AP episode were studied. Etiology, hct values at admission and at 24 h, development of severe AP according to Atlanta's criteria, pancreatic necrosis, OF and mortality were recorded. hemoconcentration was defined as hct level ＞44% for males and ＞40% for females. The t-test and χ2 test were used to assess the association of hemoconcentration to the severity, necrosis and *** accuracy was also ***: Biliary disease was the most frequent etiology(n = 148). Mean hct levels at admission were 41±6%for females and 46±7% for males (P＜0.01). Seventyeight (23%) patients had severe AP, and OF developed in 45 (13%) patients. According to contrast-enhanced computed tomography scan, 36% (54/150) patients showed pancreatic necrosis. hct levels were elevated in58% (55/96) and 61% (33/54) patients with interstitial and necrotizing pancreatitis, respectively. Neither hct levels at admission nor hemoconcentration at 24 h were associated with the severity, necrosis or OF. Sensitivity,specificity and positive predictive values for both determinations were very low; and negative predictive values were between 61% and 86%, being the highest value for ***: hct is not a useful marker to predict a worse outcome in acute pancreatitis. In spite of the high negative predictive value of hemoconcentration, the prognosis gain is limited due to an already high incidence of mild disease.

关键词：血液浓缩急性胰腺炎治疗并发症

Ivermectin Efficacy against Triatoma infestans in vivo Using hen Model

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Journal of Pharmacy and Pharmacology 2014年第6期2卷 353-358页

作者： Martin Miguel Dade Martin Rafael Daniele gustavo horacio Marin Maria Pia Silvestrini Nora Mestorino Department of Pharmacology Faculty of Medicine Sciences National University of La Plata La Plata 1900 Buenos Aires Argentina Laboratory of Pharmacological and Toxicological Studies Faculty of Veterinary Sciences National University of La Plata La Plata 1900 Buenos Aires Argentina Biostatistic Department Faculty of Veterinary Sciences National University of La Plata La Plata 1900 Buenos Aires Argentina

Chagas disease is endemic in south America. One of the major preventive actions is removing the vector TI (Triatoma infestans) from household. Although households are frequently sprayed to eliminate TI, pets and domestic animals are natural storage for vector survival. This work proposes TI elimination from farm animals using IVM (ivermectin), as a new strategy to Chagas disease control. Rhode island hens were divided into TG (treatment groups) and CG (control groups). TG received IVM while CG received DMSO. TI used in the assay was fifth instar nymphs. hens served as food source for TI. Blood was extracted for IVM chromatographic pharmacokinetic analysis. The pharmacokinetic models that best fitted the IVM plasma profiles were determined by means of the minimum Akaike information criterion estimation (MAICE) test. In TG nymphs mortality was 23.25% to 47.5% at 24-120 h, respectively. IVM plasma profile showed a first-order one-compartment model with a rapid absorption reaching a Cmax (maximum concentration) of 64.7 ± 15.4 ng/mL at 48 h (Tmax) post-application. Our data shows that TI mortality is directly related to the plasma concentration of IVM and is up to 47.5% at 120 h. Therefore, ivermectin may have a potential effect on controlling TI vector in peri-domestic level.

关键词：药剂学药理学药学药物分析药典

Recent advancements in aircraft and in situ observations of tropical cyclones

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Tropical Cyclone Research and Review 2023年第2期12卷 81-99页

作者： heather M.holbach Olivier Bousquet Lisa Bucci Paul Chang Joe Cione Sarah Ditchek Jim Doyle Jean-Philippe Duvel Jack Elston gustavo Goni Kai Kwong hon Kosuke Ito Zorana Jelenak Xiaotu Lei Rick Lumpkin Clive R.McMahon Christopher Reason Elizabeth Sanabia Lynn Keith Shay Jason A.Sippel Andrey Sushko Jie Tang Kazuhisa Tsuboki hiroyuki Yamada Jonathan Zawislak Jun A.Zhang Florida State University Northern Gulf InstituteTallahasseeUSA NOAA/OAR/Atlantic Oceanographic and Meteorological Laboratory MiamiUSA Laboratoire de L’Atmosphere et des Cyclones Sainte ClotildeFrance Institute for Coastal and Marine Research Nelson Mandela UniversityGqeberhaSouth Africa NOAA/NWS/National hurricane Center MiamiUSA NOAA/National Environmental Satellite Data and Information ServiceCollege ParkUSA Cooperative Institute for Marine and Atmospheric Studies University of MiamiMiamiFLUSA Naval Research Laboratory MontereyUSA Laboratoire de Meteorologie Dynamique ParisFrance Black Swift Technologies BoulderUSA hong Kong Observatory Hong KongChina Disaster Prevention Research Institute Kyoto UniversityGokashoJapan University of the Ryukyus OkinawaJapan University Corporation for Atmospheric Research BoulderUSA Shanghai Meteorological Service ShanghaiChina Sydney Institute of Marine Science MosmanAustralia University of Cape Town RondeboschSouth Africa United States Naval Academy AnnapolisUSA University of Miami MiamiUSA Windborne Systems Palo AltoUSA Asia-Pacific Typhoon Collaborative Research Center ShanghaiChina Shanghai Typhoon Institute CMAShanghaiChina Nagoya University NagoyaJapan Yokohama National University YokohamaJapan NOAA/OMAO/Aircraft Operations Center LakelandUSA1

Observations of tropical cyclones(TC)from aircraft and in situ platforms provide critical and unique information for analyzing and forecasting TC intensity,structure,track,and their associated *** report,prepared for the tenth International Workshop on Tropical Cyclones(IWTC-10),discusses the data collected around the world in TCs over the past four years since the IWTC-9,improvements to observing techniques,new instruments designed to achieve sustained and targeted atmospheric and oceanic observations,and select research results related to these *** the Atlantic and Eastern and Central Pacific basins,changes to operational aircraft reconnaissance are discussed along with several of the research field campaigns that have taken place *** changes in the use and impact of these aircraft observations in numerical weather prediction models are also provided along with updates on some of the experimental aircraft *** from three field campaigns in the Western Pacific basin are also *** of in-situ data collected within recent TCs such as hurricane Ian(2022),also demonstrate that new,emerging technologies and observation strategies reviewed in this report,definitely have the potential to further improve ocean-atmosphere coupled intensity forecasts.

关键词： Tropical cyclones Aircraft observations In situ observations IWTC-10