The interaction of La3+ complexes of DOTA/DTPA-glycoconjugates with the RCA120 lectin : a saturation transfer difference (STD) NMR spectroscopic study

The study of ligand-receptor interactions using high resolution NMR techniques, namely the Saturation Transfer Difference (STD), is presented for the recognition process between La(III) complexes of DOTA mono(amide) and DTPA bis(amide) glycoconjugates and the galactose specific lectin Ricinus Communis agglutinin (RCA120). This new class of Gd(III)-based potential targeted MRI contrast agents (CAs), bearing one or two terminal sugar (galactosyl or lactosyl) moieties, has been designed for in vivo binding to ASGPR (the asialoglycoprotein receptor), which is specifically expressed at the surface of liver hepatocytes, with the aim of leading to a new possible diagnosis of liver pathologies. The in vitro affinity constants of the divalent La(III)- glycoconjugate complexes to RCA120, used as a simple, water soluble receptor model, were higher than those of the monovalent analogues. The combination of the experimental data obtained from the STD NMR experiments with molecular modelling protocols (Autodock 4.1) allowed us to predict the binding mode of mono and divalent forms of these CAs to the galactose 1 binding sites of RCA120. The atomic details of the molecular interactions allowed corroborating and supporting the interaction of both the sugar moieties and the linkers with the surface of the protein and thus, their contribution to the observed interaction stabilities.Fundação para a Ciência e a Tecnologia (FCT

Mineral profiling of muscle and hepatic tissues of Australian Merino, Damara and Dorper lambs: Effect of weight loss

Seasonal weight loss (SWL) is a major constraint to extensive animal production systems. The Australian sheep production is based on merino sheep, a European breed not tolerant to SWL. Tolerant alternative breeds such as the fat-tailed Damara and the Dorper have been increasingly used in Australia and elsewhere, due to their robustness. The aim of this study was to understand the mineral profile of muscle and liver tissues of Australian Merino, Damara and Dorper, when subjected to SWL in order to understand SWL-tolerance physiology. Twenty-four lambs were divided randomly between growing (control) and nutritionally restricted groups for each breed. The trial lasted 42 days. Animals were weighed bi-weekly and at the end of the trial, lambs were slaughtered. Liver and muscle samples were taken immediately after slaughter. Mineral assessment was carried out using inductively coupled plasma–optical emission spectrometry. Analysis of variance showed mineral concentrations were generally increased in the muscle of restricted animals, mainly because of fat tissue mobilization. An increase in Zn and Fe concentrations indicates an increase of enzymatic activity in the liver of restricted sheep as well as differential abundance of Fe-containing proteins. High concentrations of Cu in the liver of Dorper indicate higher ability to accumulate this element, even under SWLinfo:eu-repo/semantics/acceptedVersio

Insights on the laccase extraction and activity in ionic-liquid-based aqueous biphasic systems

Due to their catalytic properties, selectivity, and efficiency, enzymes are excellent biocatalysts. In particular, laccases are versatile multi-copper oxidases with great interest for a wide plethora of biotechnological and environmental applications. Even though several laccase-catalysed processes have been reported at an industrial level, the high costs of their downstream processing required to provide biocatalysts with high purity levels, stability and activity remains one of the main drawbacks when economically evaluating the overall processes. Aqueous biphasic systems based on ionic liquids (ILs) can be foreseen as a promising alternative approach for the extraction and activity maintenance/improvement of enzymes, essentially due to the designer solvents ability of ionic liquids. However, to take advantage of this feature and to use the full potential of IL-based aqueous biphasic systems, it is necessary to understand the effect of ILs as phase-forming constituents and how they affect the enzymes extraction and activity. In order to overcome the lack of information on this topic in the literature, in this work, IL-based aqueous biphasic systems were investigated to extract and enhance the laccase activity, in order to gather evidences that could be used to improve the enzymes downstream processing. To this end, a wide screening of imidazolium-, pyridinium-, pyrrolidinium-, piperidinium-, tetraalkylphosphonium-, and tetraalkylammonium-based ILs as phase-forming components of ABS was carried out. Furthermore, these ILs were used to create ABS combined with salts, polymers and used as adjuvants in polymer-based ABS. Most ABS comprising ILs revealed to be highly efficient extraction platforms, allowing the complete extraction of laccase for all the conditions tested, and with an enzyme activity enhancement by more than 50%. Overall, the obtained results demonstrate that laccase preferentially partitions to the most hydrophilic phase in ABS comprising ILs, both used as adjuvants or as phase-forming components, corresponding to the phase in which the IL is enriched. Furthermore, the IL chemical structure of the IL plays a significant role in the enzyme activity, where ILs with a higher number of hydroxyl groups seem to be relevant to improve the laccase activity.publishe

A flow-through strategy using supported ionic liquids for L-asparaginase purification

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Harvest of endangered marine invertebrates in a priority area for conservation in Brazil

The harvesting of marine organisms for aquarium and ornamental art crafts is a very common and widespread activity, threatening a number of coral reef species. The goal of this manuscript is to report the intensive harvesting of two marine invertebrate species, the starfish Oreaster reticulatus and the gorgonian Phyllogorgia dilatata, in reef environments of a national priority area for the creation of a Marine Protected Area (MPA) in the Espírito Santo State, central coast of Brazil. These species were present on the 2003 Brazilian Red List of Endangered species as threatened of extinction, but P. dilatata was removed from the most recent 2014 Brazilian Red List, while O. reticulatus is currently considered vulnerable. Neither species have been assessed by IUCN. The harvest of these invertebrates was registered in 17% of the 51 days of monitoring (ten expeditions between March 2005 and February 2006), with catches of up to 24 starfishes and 1000 gorgonians in a single day. Starfishes used to be sold to local traders for US$0.50 each, and each gorgonian for US$ 0.15. This starfish is already in decline on a regional scale, and the actual harvesting rate of the gorgonian could lead to the extinction of the local population in about 17 years of exploitation. Due to the ecological importance of these two endangered species, their harvest threatens the entire local rocky reef ecosystem. Further ecological and fisheries studies on these species should be conducted, and the definition and creation of the MPA could help to manage all the unsustainable activities conducted and expected in the region

Girdling and type af cutting in rooting of jackfruit

Um experimento de enraizamento de estacas de jaqueira (Artocarpus heterophyllus Lam.) foi conduzido sob sistema de nebulização intermitente, utilizando-se um delineamento inteiramente casualizado no ananjo fatorial 3(2 x 2 x 3), onde foram estudados dois tipos de estacas (apical e basal), dois tipos de andamento dos ramos (com e sem anelamento) e três tratamentos hormonais (ácido indolbutírico a 5.000 ppm, ácido naftaleno acético a 2.000 ppm, e testemunha). A estaca do tipo basal mostrou-se superior à estaca do tipo apical quando se fez o andamento prévio dos ramos, atingindo um enraizamento de 17,8%. Quando não foi feito o anelamento dos ramos, nem a estaca basal nem a apical enraizaram. O emprego dos ácidos indoibutfrico e naftaleno acético não tiveram qualquer influência na percentagem de estacas enraizadasA branch rooting experiment was carried out with jackfruit tree (Artocarpus heterophyllus Lam.) branches under intermittent mist, using a completely randomized block design with a factorial 3(2 x 2 x 3) arrangement. Two cutting types (terminal an subterminal, two girdling types (with and without girdling) and three hormone treatments (indolbutyric acid 5.000 ppm, naphthaleneacetic acid 2.000 ppm, and check) were used: The subterminal cutting proved to be superior to the terminal, and reached 17,8% of rooting when girdling was done. However, when branches were left ungirdled, neither terminal nor subterminal cutting took root. The use of indolbutyric and naphthaleneacetic acids did not improve rooting percentage

Immobilization of L-asparaginase towards surface-modified carbon nanotubes

L-asparaginase (ASNase, EC 3.5.1.1) is an enzyme that catalyzes L-asparagine hydrolysis into L-aspartic acid and ammonia and is mainly applied in pharmaceutical and food industries [1]. The ASNase currently commercialized for pharmaceutical purposes is produced from two main bacterial sources: recombinant Escherichia coli and Erwinia chrysanthemi. However, some disadvantages are associated with its free form, such as the shorter half-life [2]. Immobilization of ASNase has been proposed as an efficient approach to overcome this limitation [3]. In this work, a straightforward method, including the functionalization of multi-walled carbon nanotubes (MWCNTs) through a hydrothermal oxidation treatment with nitric acid, and the immobilization of ASNase by adsorption over pristine and modified MWCNTs was investigated. Different operation conditions, including pH, contact time, ASNase/MWCNT mass ratio, and the operational stability of the immobilized ASNase were evaluated. The characterization of the ASNase-MWCNT bioconjugate was addressed using different techniques, namely Transmission Electron Microscopy (TEM), Thermogravimetric analysis (TGA), and Raman spectroscopy. Functionalized MWCNTs showed promising results, with an immobilization yield and a relative recovered activity of commercial ASNase above 95%, under the optimized adsorption conditions (pH 8, 60 min of contact and 1.5´10–3 g.mL-1of ASNase). The ASNase-MWCNT bioconjugate also showed improved enzyme operational stability (6 consecutive reaction cycles without activity loss), proving its suitability for application in industrial processes.publishe

Immobilization of L-asparaginase towards surface-modified carbon nanotubes

L-asparaginase (LA) is an enzyme that catalyzes L-asparagine hydrolysis into L-aspartic acid and ammonia and is mainly applied in pharmaceutical and food industries. The LA currently commercialized for pharmaceutical purposes is produced from two main bacterial sources: recombinant Escherichia coli and Erwinia chrysanthemi. However, some disadvantages are associated with its free form, such as the shorter half-life. Immobilization of LA has been proposed as an efficient approach to overcome this limitation. In this work, a straightforward method, including the functionalization of multi-walled carbon nanotubes (MWCNTs) through a hydrothermal oxidation treatment and the immobilization of LA by adsorption over pristine and modified MWCNTs was investigated. Different operation conditions, including pH, contact time, ASNase/MWCNT mass ratio, and the operational stability of the immobilized LA, were evaluated. The characterization of the LA-MWCNT bioconjugate was addressed using different techniques, namely Transmission Electron Microscopy (TEM), Thermogravimetric analysis (TGA), and Raman spectroscopy. Functionalized MWCNTs showed promising results, with an immobilization yield and a relative recovered activity of commercial LA above 95%, under the optimized adsorption conditions (pH 8, 60 min of contact, and 1.510–3 g.mL-1 of LA). The LA-MWCNT bioconjugate also showed improved enzyme operational stability (6 consecutive reaction cycles without activity loss), proving its suitability for application in industrial processes.publishe

Hydrological influence on the evolution of a subtropical mangrove ecosystem during the late Holocene from Babitonga Bay, Brazil

Mangroves are key ecosystems which respond to global changes in tropical and subtropical regions worldwide. We describe late Holocene mangroves that established close to the southernmost limit (28°S) for this type of ecosystem in South America. Our findings are based on a C dated core obtained from Babitonga Bay, Santa Catarina State, Brazil (26°12′S, 48°33′W). Analysis of palynology, sedimentary facies, isotopic and elemental data shows that mangrove establishment took place ~500 yrs. B.C.E., following an increase in humidity, and expanded further during the Roman Warm Period and at the end of Dark Age Cold Period. Mangrove and precipitation proxies records appear to be sensitive to rainfall patterns imposed both by the expansion/retraction of the Intertropical Convergence Zone and also the interaction with the South Atlantic Subtropical Anticyclone which affects coastal region due to sea surface temperature variations.The authors thank the Coastal Dynamic Laboratory (LADIC-UFPA) , C-14 Laboratory of the Center for Nuclear Energy in Agriculture (CENA-USP) , University of Joinville (UNIVILLE) and Radiocarbon Laboratory (LAC-UFF) for all infrastructure and support. We also thank three anonymous Reviewers and Prof. H. Falcon-Lang for their constructive comments. The first and third author thanks Brazilian Council for Technology and Science-CNPq for fellowship (process 131813/2016-1 , 165911/2015-8 and 305074/2017-2 ). This study was financed by CNPq ( 445111/2014-3 , 405060/2013- 0 ) and FAPESP ( 2011/00995-7 , 2017/03304-1, and 2020/13715-1 ). This study also was financed in part by the Coordenação de Aperfeiçoamento de Pessoal Nível Superior – Brazil (CAPES) – Finance Code 001