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We carried out a genome-wide association study in Latin Americans and identified novel face morphology loci.., To characterize the genetic basis of facial features in Latin Americans, we performed a genome-wide association study (GWAS) of more than 6000 individuals using 59 landmark-based measurements from two-dimensional profile photographs and ~9,000,000 genotyped or imputed single-nucleotide polymorphisms. We detected significant association of 32 traits with at least 1 (and up to 6) of 32 different genomic regions, more than doubling the number of robustly associated face morphology loci reported until now (from 11 to 23). These GWAS hits are strongly enriched in regulatory sequences active specifically during craniofacial development. The associated region in 1p12 includes a tract of archaic adaptive introgression, with a Denisovan haplotype common in Native Americans affecting particularly lip thickness. Among the nine previously unidentified face morphology loci we identified is the VPS13B gene region, and we show that variants in this region also affect midfacial morphology in mice.

Background : Squamous cell carcinoma (SCC) is one of the most common malignant skin tumors in domestic animals. Histologically, they are characterized by a proliferation of neoplastic keratinocytes with varied keratin production. Some SCCs have peculiar histological characteristics that permit them to be classified into uncommon to rare histological subtypes, reported in animals and humans. However, according to the authors' knowledge, the mucin-producing subtype described in humans has not yet been reported in animals. In this study, we report the occurrence of two mucin-producing SCCs in dogs, a histological presentation similar to that seen in cutaneous SCCs with mucinous metaplasia in humans. Cases : Two dogs, a 5-year-old Yorkshire female and a 17-year-old Dachshund male, had a skin nodule near the tail and on the right eyelid. The nodules varied from 1 to 5 cm in diameter, were firm and covered with skin and hair. The cut surface was firm and white. Histological findings were compatible with squamous cell carcinoma, characterized by a neoplastic proliferation of keratinocytes originating in the epidermis and infiltrating the dermis. The keratinocytes were arranged in islands and occasional anastomosed cords, supported by a fibrous stroma. The formation of pearls varied from moderate to sparse. The nuclear and cellular pleomorphism was accentuated in case two and moderate in case one. Mitosis figures ranged from two to five in a high magnification field. Within the neoplasm, there were large vacuolated neoplastic cells with slightly fibrillar intracytoplasmic basophilic content. This content has been rarely observed in an extracellular medium. The presence of mucin was confirmed by positive Alcian Blue (AA) staining. In immunohistochemistry (IHC), tumor cells showed strong immunostaining for pancitokeratin, and in areas with marked mucin deposition, immunostaining was predominantly moderate to weak. No tumor cells were immunostained for CD34 and Blc-2 antibodies. Compared to AA and Harris' hematoxylin, it was possible to demonstrate the presence of mucin in the cytoplasm of neoplastic keratinocytes using IHC. No vascular or lymphatic invasion by neoplastic cells was observed. The average cell proliferation index assessed by counting the nucleolar argyrophilic organizing regions (AgNOR) was 3.4 in case 1 and 4.5 in case 2. Discussion : Although the SCC routinely does not present a diagnostic challenge in veterinary practice, the histological presentation of the reported cases does not fit the current classification available in veterinary medicine. The histological presentation observed in these two dogs is similar to that described for cutaneous SCCs with mucinous metaplasia in humans, and so far not described in animals. The observation of intracytoplasmic mucin in humans is an essential finding for the diagnosis of SCC with mucin metaplasia. In the present cases, we observed a slightly basophilic amorphous substance in the cytoplasm of proliferated neoplastic keratinocytes, which stained strongly in blue when applied the Alcian Blue (AA) histochemical technique. This observation became more evident when using IHC counterstained with AA and Harris hematoxylin . In the histological analysis, the absence of an adenoid growth pattern or glandular formation amid neoplastic proliferation ruled out the possibility of a mucinous adenocarcinoma or a mucoepidermoid carcinoma. In addition, we could rule out a follicular neoplasia, including keratinizing infundibular acanthoma due to the absence of a central pore and the absence of immunostaining for CD34 and Bcl-2. These immunohistochemical findings, together with histological findings, reinforce the diagnosis of SCC with mucinous metaplasia in our dogs.

Background : Pythiosis is an infectious disease caused by the oomycete Pythium insidiosum , with higher occurrence in wetlands and hot climate regions. This microorganism develops its cycle in aquatic plants, and most cases happen because of the contact of animals or people with water containing the motile zoospores (infectious form). Horses are the principal species affected and develop principally cutaneous and subcutaneous lesions, but the gastrointestinal tract is seldom affected. Humans develop various forms of pythiosis, such as a vascular form. The objectives of the current study are to describe an unusual case of intestinal pythiosis, its clinical signs, aspects of pathogenesis, and diagnosis. Case : A 13-year-old Crioula mare, from Santa Maria, RS, Brazil, presented with reduced food and water intake, apathy, restlessness, rolling, nasal reflux, firm abdomen upon palpation, and tachypnea during 2 days. The horses of this farm were fed native pasture and horse feed, and they had access to a nearby pond. Two days following the start of the clinical signs, the horse died and was necropsied on the farm. During necropsy, there was around 400 mL of reddish effusion in the abdominal cavity (modified transudate). A 15 cm segment of jejunum was firm upon palpation and had a severe transmural thickening. The wall of the affected area was up to 3 cm in thickness and firm, with small yellowish and irregular masses that stood out and looked friable, interpreted as kunkers. Microscopically, the yellowish masses (kunkers) were characterized by dense accumulations of intact and degenerate eosinophils (eosinophilic necrosis). Within these kunkers, and also on their periphery, there were multiple negatively stained hyphal profiles. Hyphae were also seen on the wall of small arteries inside the kunkers. These hyphae had nearly parallel walls and were occasionally septate, with a diameter of about 10 μm. The hyphae stained black with Grocott’s methenamine silver stain (GMS) and were positive on immunohistochemistry (IHC) using specific anti- P. insidiosum polyclonal antibody. Discussion : Colic is the principal clinical presentation in horses suffering from gastrointestinal tract disorders. In spite of the presence of colic in this horse, intestinal pythiosis was not clinically suspected in this case, principally because of its rare occurrence in horses. Even though it is persistent to observe horses ingesting water with suspected contamination by P. insidiosum , the cutaneous form of pythiosis is a lot more common than the intestinal form. This is the only case of intestinal pythiosis in a horse in 52 years of routine diagnosis in our laboratory. It is suspected that this horse got infected by drinking contaminated water from the nearby pond and microlesions in the intestinal mucosa due to plant material or some unknown pathogen may have favored zoospore adhesion, encysting and starting the colonization of the tissue by emitting a germ tube. A macroscopic observation of kunkers in the intestine is rare, but when observed it is highly indicative of pythiosis. The immunohistochemistry technique using anti- P. insidiosum antibody, in accordance with the literature, validates the diagnosis of pythiosis. In this case, the presence of hyphae in the blood vessel wall inside the kunkers propose that this finding may be involved in the spread of the lesion but needs more detailed studies.

Exposure to high concentrations of cadmium (Cd), widely used in many industries and found in air, food and contaminated water, is not uncommon. Cd damages the cardiovascular system, but the vascular mechanisms involved are not fully understood. This study investigated the mechanisms involved in cardiovascular damage after exposure to high Cd concentrations. Three-month-old male Wistar rats were treated intraperitoneally for 14 days with distilled water (Untreated group) or 1 mg/kg cadmium chloride (Cd group). We investigated the systolic blood pressure (SBP) and vascular reactivity of mesenteric resistance arteries (MRA) and the aorta by analysing contractile and relaxation responses in the absence and presence of the endothelium; we also evaluated pathways involved in vascular tone regulation. Superoxide anion production, COX-2 protein expression and in situ detection of COX-2, AT-1, and NOX-1 were evaluated. Oxidative status, creatinine level and angiotensin-converting enzyme (ACE) activity in plasma were also evaluated. Fourteen-day exposure to a high Cd concentration induced hypertension associated with vascular dysfunction in MRA and the aorta. In both vessels, there was increased participation of cyclooxygenase 2 (COX2), angiotensin II type 1 (AT1) receptor and NOX1. MRA also presented endothelial dysfunction, denoted by impaired acetylcholine-mediated relaxation. All vascular changes were accompanied by increased reactive oxygen species production and COX2, NOX1 and AT1 receptor expression in vascular tissue. Overall, high Cd concentrations induced cardiovascular damage: hypertension, endothelial dysfunction and vascular damage in conductance and resistance arteries, NADPH oxidase, renin–angiotensin system and COX2 pathway activation., Supported by National Council for Scientific and Technological Development – CNPq [CNPq 307399/2017-6]; Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) Fundação de Amparo à Pesquisa do Rio Grande do Sul - FAPERGS/Brazil [PQG:19/2551-0001810-0]; Programa Nacional de Cooperação Acadêmica; Pró-reitoria de Pesquisa - Universidade Federal do Pampa [Nº 20180615102630]; FAPES/CNPq/PRONEX [Nº 80598773] and Spanish Goverment by the Agencia Estatal de Investigación (AEI) and Fondo Europeo de Desarrollo Regional (FEDER) [AGL2017-89213]. JEGPJr and PZM were supported by CAPES/Brazil, MDR by FAPERGS/Brazil (Fundação de Amparo à Pesquisa do Rio Grande do Sul).

12 páginas.- 5 figuras.- 4 tablas.- 103 referencias.- Supplementary data to this article can be found online at https://doi.org/10.1016/j.apenergy.2020.114493.0.00.51.01.52.0, Novel laccases have promising and valuable applications in biorefineries. This investigation documents, for the first time, the potential of depolymerising and repolymerising lignin by the secretome, rich in laccases, from a newly isolated white-rot basidiomycete Marasmiellus palmivorus VE111, for further saccharification and ethanolic fermentation steps. Proteomic analyses of the secretome of M. palmivorus show that laccases are the most predominant enzyme released by this fungus. The whole crude enzymatic broth is used for the delignification of lignin in Eucalyptus globulus wood, with the aim of enhancing the saccharification by cellulolytic and xylanolytic enzymes from Penicillium echinulatum S1M29. In addition, two different strategies, namely, laccase treatment before and after enzymatic hydrolysis, are employed to detoxify steam-exploded E. globulus wood. The objective is to increase the fermentative performance by removing substances formed during the feedstock pretreatment that can inhibit microbial fermentation. The E. globulus wood delignification results in a 31% decrease in the lignin content and a 10% increase in the glucose yield after hydrolysis. An important finding of the present work is the successful wood delignification in the absence of laccase mediators. This laccase-rich preparation also demonstrates its potential in removing the phenolic inhibitors present in steam-exploded E. globulus wood, increasing the ethanol yield by an additional 10%. Furthermore, it is important to highlight that these findings are achieved in the absence of commercial enzymes, making M. palmivorus laccases a potential candidate not only for the production of biofuels but also for the generation of lignin-derived aromatic compounds for different applications in the biotechnology industry. © 2020 Elsevier Ltd, Funding from University of Caxias do Sul (UCS) Coordination of Improvement of Higher Education Personnel (CAPES) (Process 3255/2013), Research Support Foundation of the State of Rio Grande do Sul (Process 16/2551-0000 246-0), Brazilian Agricultural Research Corporation (EMBRAPA) (34/2014), National Council for Scientific and Technological Development (CNPq) (Process 201925/2018-4), the Biobased Industries Joint Undertaking (www.bbi-europe.eu) project EnzOx2 (H2020-BBI-PPP-2015-2-720297, www.enzox2.eu), the projects RETOPROSOST-2-CM [S2018/EMT-4459] financed by the Comunidad de Madrid, HIDROFUN [BIO2015-68387-R], and GLYSUS [RTI2018-093683-B-I00] from MINECO/FEDER, and the CSIC Project (201740E07)

15 Páginas.-- 5 Tablas.-- 4 Figuras, Different cultivation strategies have been developed with the aim of increasing the production rate of microalgal pigments. Specifically, biotechnological approaches are designed to increase antioxidant metabolites as chlorophyll and carotenoids. However, although significant advances have been built up, available information regarding both the chlorophyll metabolism and their oxidative reactions in photobioreactors is scarce. To unravel such processes, the detailed chlorophyll and carotenoid fraction of Scenedesmus obliquus has been studied by HPLC-ESI/APCI-hrTOF-MS from phototrophic and heterotrophic cultures. Scenedesmus is provided with a controlled strategy of interconversion between chlorophyll a and b to avoid the formation of reactive oxygen species (ROS) at high irradiances in addition to the photoacclimation of carotenoids. Indeed, precise kinetics of 132-hydroxy- and 151-hydroxy-lactone chlorophyll metabolites shows the existence of a chlorophyll oxidative metabolism as a tool to manage the excess of energy at high light conditions. Unexpectedly, the oxidation under phototrophy favored chlorophyll b metabolites over the chlorophyll a series, while the heterotrophic conditions exclusively induced the formation of 132-hydroxy-chlorophyll a. In parallel, during the first 48 h of growth in the dark, the chlorophyll fraction maintained a promising steady state. Although future studies are required to resolve the biochemical reactions implied in the chlorophyll oxidative metabolism, the present results agree with phytoplankton metabolism., This work was supported by the Ministerio de Ciencia, Investigación y Universidades, Agencia Estatal de Investigación y Fondo Europeo de Desarrollo Regional (FEDER), grant number RTI2018-095415-B-I00. MM was supported with a fellowship from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001 and the Brazilian Funding Agency FAPERGS (Fundação de Amparo a pesquisa do estado do Rio Grande do Sul). The APC was partially funded by CSIC.

International audience; Medical devices (indwelling) have greatly improved healthcare. Nevertheless, infections related to the use of these apparatuses continue to be a major clinical concern. Biofilms form on surfaces after bacterial adhesion, and they function as bacterial reservoirs and as resistance and tolerance factors against antibiotics and the host immune response. Technological strategies to control biofilms and bacterial adhesion, such as the use of surface coatings, are being explored more frequently, and natural peptides may promote their development. In this study, we purified and identified antibiofilm peptides from Capsicum baccatum (red pepper) using chromatography-tandem mass spectrometry, MALDI-MS, MS/MS and bioinformatics. These peptides strongly controlled biofilm formation by Staphylococcus epidermidis, the most prevalent pathogen in device-related infections, without any antibiotic activity. Furthermore, natural peptide-coated surfaces dislayed effective antiadhesive proprieties and showed no cytotoxic effects against different representative human cell lines. Finally, we determined the lead peptide predicted by Mascot and identified CSP37, which may be useful as a prime structure for the design of new antibiofilm agents. Together, these results shed light on natural Capsicum peptides as a possible antiadhesive coat to prevent medical device colonization.

Novozym 435 (N435) is a commercially available immobilized lipase produced by Novozymes. It is based on immobilization via interfacial activation of lipase B from Candida antarctica on a resin, Lewatit VP OC 1600. This resin is a macroporous support formed by poly(methyl methacrylate) crosslinked with divinylbenzene. N435 is perhaps the most widely used commercial biocatalyst in both academy and industry. Here, we review some of the success stories of N435 (in chemistry, energy and lipid manipulation), but we focus on some of the problems that the use of this biocatalyst may generate. Some of these problems are just based on the mechanism of immobilization (interfacial activation) that may facilitate enzyme desorption under certain conditions. Other problems are specific to the support: Mechanical fragility, moderate hydrophilicity that permits the accumulation of hydrophilic compounds (e.g., water or glycerin) and the most critical one, support dissolution in some organic media. Finally, some solutions (N435 coating with silicone, enzyme physical or chemical crosslinking, and use of alternative supports) are proposed. However, the N435 history, even with these problems, may continue in the coming future due to its very good properties if some simpler alternative biocatalysts are not developed., We gratefully recognize the financial support from MINECO from the Spanish Government (project number CTQ2017-86170-R, Colciencias, Ministerio de Educación Nacional, Ministerio de Industria, Comercio y Turismo e ICETEX, Convocatoria Ecosistema Científico – Colombia Científica. Fondo Francisco José de Caldas, Contrato RC-FP44842-212-2018 and Colciencias (Colombia) (project number FP44842-076-2016), Generalitat Valenciana (PROMETEO/2018/076), FAPERGS (project number 17/2551-0000939-8), CONICET (R. Argentina), FUNCAP (project number BP3-0139-00005.01.00/18) and ANPCyT (PICT 2015-0932 and PICT CABBIO 4687)., We acknowledge support of the publication fee by the CSIC Open Access Support Initiative through its Unit of Information Resources for Research (URICI)

The increase in life expectancy of the world population is associated with a higher prevalence of neurodegenerative diseases. Alzheimer's Disease (AD) is the most common neurodegenerative disease, affecting currently 43 million people over the world. To date, most of the pharmacological interventions in AD are intended for the alleviation of some of its symptoms, and there are no effective treatments to inhibit the progression of the disease. Translocator protein (TSPO) is present in contact points between the outer and the inner mitochondrial membranes and is involved in the control of steroidogenesis, inflammation and apoptosis. In the last decade, studies have shown that TSPO ligands present neuroprotective effects in different experimental models of AD, both in vitro and in vivo. The aim of this review is to analyze the data provided by these studies and to discuss if TSPO could be a viable therapeutic target for the development of new treatments for AD., This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq – Chamada Universal 429639/2018-0) and the Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS – APE 18/2551-00011058-8). Maria Flávia Ribeiro received a CNPq 1D Researcher Productivity Grant. The authors thank Gabrielli Tiburi Soares Pires for helping in the preparation of the figure.

This study evaluated the effect of protein hydrolysate (PH) and clove essential oil (CEO) on agar film properties and the shelf-life improvement of flounder fillets. Firstly, Argentine croaker muscle protein was alkali-solubilised and recovered to obtain a stable substrate. This substrate was hydrolysed to 20% degree of hydrolysis, by Alcalase (A20) and Protamex (P20). The molecular weight (MW) distribution, amino acid and in vitro antimicrobial activity of these hydrolysates were tested. Agar films were prepared with and without (control) incorporation of PH (A20 hydrolysate) or CEO and characterised by mechanical properties, water vapour permeability, solubility and optical properties. The microbiological, physical and chemical properties of flounder fillets coated with these films were monitored during storage at 5 °C for 15 days. Compared to P20, A20 hydrolysate possessed lower MW, higher hydrophobic amino acid content and the most effective antimicrobial activity against selected pathogens, such as Staphylococcus aureus, Yersinia enterocolitica, Aeromonas hydrophila, Debaryomyces hansenii and Listeria innocua. The presence of PH increased the water solubility, water vapour permeability, elongation at break and yellowness of the films. The CEO films possessed less transparency than PH films and control films, due to the presence of oil in the formulations. Fillets covered with PH and CEO films, especially the latter, presented lower total volatile bases and pH values, and delayed considerably the growth of HS-producing microorganisms compared to the control films. The CEO might be used as a natural biopreservative to extend the flounder shelf-life. However, further studies are necessary, to assess the antimicrobial effect of PH when used as food packaging., The authors acknowledge the financial support provided by the Brazilian Scientific and Technological National Council (CNPq), a grant from the Rio Grande do Sul State Research Foundation (FAPERGS) and the Spanish Ministry of Economy and Competitiveness through projects AGL2011-27607 and AGL2014-52825-R.

In this work, the combined use of ultrasound energy and molecular sieves was investigated for the synthesis of ethyl butyrate, ester with mango and banana notes, catalyzed by the immobilized lipase from Thermomyces lanuginosus (Lipozyme TL-IM). Initially, the best concentrations of biocatalysts (35%) and butyric acid (0.7 M) were tested using ultrasound as an alternative to mechanical agitation. The amount of acid in the reaction could be increased by 2-fold when compared to previous works where mechanical agitation was used. In the next step, substrate molar ratio and reaction temperature were optimized and the best conditions were at their lowest levels: 1:1 (acid:alcohol), and 30 °C, reaching 61% of conversion in 6 h. Molecular sieves (3 Å) were added to optimized reaction medium in order to remove the formed water and improve the maximum yield. The reaction yield increased 1.5 times, reaching 90% of conversion in 6 h, when 60 mg of molecular sieves per mmol of butyric acid was used. Finally, the reuse of Lipozyme TL-IM for the ultrasound-assisted synthesis of ethyl butyrate was verified for 10 batches, without any appreciable loss of activity, whereas in systems using mechanical agitation, the biocatalyst was completely inactivated after 5 batches. These results suggest that the combined use of ultrasound and molecular sieves greatly improve esterification reactions by stabilizing the enzyme and increasing yields., This work was supported by grants and scholarships (first and second authors) from FAPERGS (Fundação de Amparo a Pesquisa do Rio Grande do Sul – ARD/2011), CNPq (Brazilian Bureau of Science and Technology), and CTQ2009-07568 (Ministerio de Ciencia y Innovación, Spain).

The concept of the combi-lipase biocatalyst has been proposed. It is based on the combination of different lipases as biocatalysts in reactions using heterogeneous substrates. The hydrolysis of soybean oil was evaluated as a model substrate, and Novozym 435 (CALB), Lipozyme TL-IM (TLL), and Lipozyme RM-IM (RML) were used as biocatalysts. Results showed that, although individually TLL was the most active enzyme, whereas CALB was the less active one, the combination of 80% of RML and 20% of CALB was the best biocatalyst. Reaction parameters were optimized, allowing to obtain more than 80% hydrolysis in 24 h using the combi-lipase, up from less than 50% with any individual lipase. Reusability of the combi-lipase showed that it could be used for at least 15 cycles without any significant decrease. The concept of the combi-biocatalyst might be a useful technology for reactions including full modification of heterogeneous substrates., This work was supported by grants from Fundaçao de Amparo a Pesquisa do Rio Grande do Sul (FAPERGS; ARD/2011, Brazil), from CNPq (Brazilian Bureau of Science and Technology), and CTQ2009-07568 from Spanish Ministerio de Ciencia e Innovación.

Butyl butyrate is an ester present in pineapple flavor, which is very important for the food and beverages industries. In this work, the optimization of the reaction of butyl butyrate synthesis catalyzed by the immobilized lipase Lipozyme TL-IM was performed. n-Hexane was selected as the most appropriate solvent. Other reaction parameters such as temperature, substrate molar ratio, biocatalyst content and added water, and their responses measured as yield, were evaluated using a fractional factorial design, followed by a central composite design (CCD) and response surface methodology. In the fractional design 24-1, the four variables were tested and temperature and biocatalyst content were statistically significant and then used for optimization on CCD. The optimal conditions for butyl butyrate synthesis were found to be 48°C; substrate molar ratio 3:1 (butanol:butyric acid); biocatalyst content of 40% of acid mass. Under these conditions, over 90% of yield was obtained in 2 h. Enzyme reuse was tested by washing the biocatalyst with n-hexane or by direct reuse. The direct reuse produced a rapid decrease on enzyme activity, while washing with n-hexane allowed reusing the enzyme for five reactions cycles keeping approximately 85% of its activity. © 2013 American Institute of Chemical Engineers., This work was supported by grants from Fundação de Amparo a Pesquisa do Rio Grande do Sul (FAPERGS–ARD/2011) from CNPq (Brazilian Bureau of Science and Technology), and CTQ2009‐07568 from Spanish Ministerio de Ciencia e Innovación. The authors would like to thank Mr. Ramiro Martínez (Novozymes, Spain) for kindly supplying the enzymes used in this research. They also thank Ministerio de Ciencia e Innovación by a fellowship for C. Garcia‐Galan and CNPq–Brazil for a fellowship to J. L. R. Friedrich and FAPERGS–Brazil for a fellowship to A. B. Martins.

Schistosomiasis is a chronic parasitic disease of humans, with two species primarily causing the intestinal infection: Schistosoma mansoni and Schistosoma japonicum. Traditionally, diagnosis of schistosomiasis is achieved through direct visualisation of eggs in faeces using techniques that lack the sensitivity required to detect all infections, especially in areas of low endemicity. A recently developed method termed Helmintex™ is a very sensitive technique for detection of Schistosoma eggs and exhibits 100% sensitivity at 1.3 eggs per gram of faeces, enough to detect even low-level infections. The Helminthex™ method is based on the interaction of magnetic microspheres and schistosome eggs. Further understanding the underlying egg-microsphere interactions would enable a targeted optimisation of egg-particle binding and may thus enable a significant improvement of the Helmintex™ method and diagnostic sensitivity in areas with low infection rates. We investigated the magnetic properties of S. mansoni and S. japonicum eggs and their interactions with microspheres with different magnetic properties and surface functionalization. Eggs of both species exhibited higher binding affinity to the magnetic microspheres than the non-magnetic microspheres. Binding efficiency was further enhanced if the particles were coated with streptavidin. Schistosoma japonicum eggs bound more microspheres compared with S. mansoni. However, distinct differences within eggs of each species were also observed when the distribution of the number of microspheres bound per egg was modelled with double Poisson distributions. Using this approach, both S. japonicum and S. mansoni eggs fell into two groups, one having greater affinity for magnetic microspheres than the other, indicating that not all eggs of a species exhibit the same binding affinity. Our observations suggest that interaction between the microspheres and eggs is more likely to be related to surface charge-based electrostatic interactions between eggs and magnetic iron oxide rather than through a direct magnetic interaction., This work was funded by grants from the Australian Research Council (ARC) and National Health and Medical Research Council (NHMRC) of Australia, a University of Western Australia/University of Queensland (Australia) Bilateral Research Collaboration Award as well as grants from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and the Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS) in Brazil. Travel to Australia by RRFC was funded by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil. SK is funded by an NHMRC Early Career Fellowship (GNT 1052760). LG is the beneficiary of a post-doctoral grant from the AXA Research Fund (Europe).

Lipase from Burkholderia cepacia (PS) has been immobilized on Accurel MP 1000, and their performance has been compared to that of the most widely used immobilized commercial preparation of this enzyme. The maximum loading was 18 mg protein/g support, and its thermal and solvent stability was much higher than that of the commercial. PS preparations were used in hydrolysis of triacetin and methyl mandelate, in the esterification of oleic acid and ethanol and in the kinetic resolution of 1,3,4-tri-O-benzyl-myo-inositol (DL-1) using vinylacetate as activated acyl donor. For all reactions studied, PS on Accurel was more active than PS-IM. The conversion in the kinetic resolution of racemic DL-1 was optimized using response surface methodology. Optimal conditions were determined to be 2.0 mg/mL of substrate, temperature of 40 °C, 2.0 mL of vinyl acetate and without water addition. Under these conditions, maximum loaded Accurel-PS preparation permitted to improve the activity in this kinetic resolution compared to the PS commercial preparation by a 55-fold factor, and compared to Novozym 435 (the most active described in literature for this reaction) by a 23-fold factor. The conversion attained was 49.9% ± 0.3 of conversion and ee of 99% after 24 h. The reusability studies showed maintenance of conversion and ee during eight cycles., We gratefully recognize the support from the MINECO of Spanish Government, by the grant CTQ2013-41507-R, and Brazilian FAPERJ, CNPq and CAPES for funding and/or fellowships, including a PVE project for R. Fernandez-Lafuente.

Aulacothrips Hood, 1952 is a remarkable South American genus of ectoparasitic thrips which feed on gregarious ant-tended hemipterans (Hemiptera: Auchenorrhyncha). Here we describe two new species of this genus infesting membracid treehoppers in Colombia and Brazilian Amazon, and provide a key to the five Aulacothrips spp. now recognised. Aulacothrips levinotus n. sp. is unique in having pronotum with only two pairs of major setae; and Aulacothrips tenuis n. sp. has unusually long abdominal tergite IX. These findings will be helpful for understanding the species diversification in this genus, bringing a new perspective on the diversity of Aulacothrips spp. and their myrmecophilous hemipteran hosts. © 2014, Springer Science+Business Media Dordrecht., Financial support was provided by the Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul. LAK was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP 10/51340-8) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES 3200-14-0).

This study investigated the effects of prolonged exposure of silver catfish (Rhamdia quelen) to the essential oil (EO) of Hesperozygis ringens. Ventilatory rate (VR), stress and metabolic indicators, energy enzyme activities, and mRNA expression of adenohypophyseal hormones were examined in specimens that were exposed for 6 h to 0 (control), 30 or 50 µL L−1 EO of H. ringens in water. Reduction in VR was observed in response to each treatment, but no differences were found between treatments. Plasma glucose, protein, and osmolality increased in fish exposed to 50 µL L−1. Moreover, lactate levels increased after exposure to both EO concentrations. Plasma cortisol levels were not changed by EO exposure. Fish exposed to 30 µL L−1 EO exhibited higher glycerol-3-phosphate dehydrogenase (G3PDH) activity, while exposure to 50 µL L−1 EO elicited an increase in glucose-6-phosphate dehydrogenase (G6PDH), fructose-biphosphatase (FBP), and 3-hydroxyacyl-CoA-dehydrogenase (HOAD) activities compared with the control group. Expression of growth hormone (GH) only decreased in fish exposed to 50 µL L−1 EO, while somatolactin (SL) expression decreased in fish exposed to both concentrations of EO. Exposure to EO did not change prolactin expression. The results indicate that GH and SL are associated with energy reorganization in silver catfish. Fish were only slightly affected by 30 µL L−1 EO of H. ringens, suggesting that it could be used in practices where a reduction in the movement of fish for prolonged periods is beneficial, i.e., such as during fish transportation., This study was supported by research funds from the Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS/PRONEX), Ministério da Pesca/Ministério da Ciência e Tecnologia/FINEP, Conselho Nacional de Pesquisa e Desenvolvimento Científico (CNPq) and by project AGL2013-48835-C2-1-R (Ministerio de Economía y Competividad) to J.M. Mancera (Spain). B. Baldisserotto received a research fellowship from CNPq. C. Toni and C.G. Pinheiro received PhD fellowships from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and J.A. Martos-Sitcha is supported by a PhD fellowship (FPU, Reference AP2008-01194) from Ministerio de Educación (Spain).

In this work, it is compared the performance of three commercial lipase preparations (Novozym 435, Lipozyme TL-IM, and Lipozyme RM-IM) in the synthesis of flavor esters obtained by esterification of acetic, propionic, and butyric acids using ethanol, isopropyl alcohol, butanol, or pentanol. A comprehensive comparison was performed verifying activities of these three enzyme preparations versus the different couples of substrates, checking the obtained yields. In general, the longer the acid chain, the higher the reaction yields. Novozym 435 was the most efficient enzyme in most cases, and only Lipozyme RM-IM offered better results than Novozym 435 in the production of ethyl butyrate. Reactions with butyric acid showed the highest conversion rates using all biocatalysts. Using optimal substrates, the reactions catalyzed by the three enzymes were optimized using the response surface methodology, and the catalytic performance of the biocatalysts in repeated batches was assessed. After optimization, yields higher than 90% were obtained for all three enzymes, but Lipozyme TL-IM needed four-times more biocatalyst content than the other two preparations. Novozym 435 kept over 80% of its activity when reused in 9 successive batches, whereas Lipozyme RM-IM can be reused 5 times and Lipozyme TL-IM only 3 times. In general, Novozym 435 showed to be more suitable for these reactions than the other two enzyme preparations., This work was supported by grants and scholarships (first and second authors) from FAPERGS (Fundação de Amparo a Pesquisa do Rio Grande do Sul process 11/0485-0 ARD/2011), CNPq (Brazilian Bureau of Science and Technology process 478169/2011-6 Universal 2011), and CTQ2009-07568 (Ministerio de Ciencia y Innovación, Spain).

The influence of low-frequency ultrasound (40 kHz) in the esterification reaction between acetic acid and butanol for flavor ester synthesis catalyzed by the commercial immobilized lipase B from Candida antarctica (Novozym 435) was evaluated. A central composite design and the response surface methodology were used to analyze the effects of the reaction parameters (temperature, substrate molar ratio, enzyme content and added water) and their response (yields of conversion in 2.5 h of reaction). The reaction was carried out using n-hexane as solvent. The optimal conditions for ultrasound-assisted butyl acetate synthesis were found to be: temperature of 46 °C; substrate molar ratio of 3.6:1 butanol:acetic acid; enzyme content of 7%; added water of 0.25%, conditions that are slightly different from those found using mechanical mixing. Over 94% of conversion was obtained in 2.5 h under these conditions. The optimal acid concentration for the reaction was determined to be 2.0 M, compared to 0.3 M without ultrasound treatment. Enzyme productivity was significantly improved to around 7.5-fold for each batch when comparing ultrasound and standard mechanical agitation. The biocatalyst could be directly reused for 14 reactions cycles keeping around 70% of its original activity, while activity was virtually zeroed in the third cycle using the standard mixing system. Thus, compared to the traditional mechanical agitation, ultrasound technology not only improves the process productivity, but also enhances enzyme recycling and stability in the presence of acetic acid, being a powerful tool to improve biocatalyst performance in this type of reaction., This work was supported by grants from Fundação de Amparo a Pesquisa do Rio Grande do Sul (FAPERGS – ARD/2011), from CNPq (Brazilian Bureau of Science and Technology), and CTQ2009-07568 from Spanish Ministerio de Ciencia e Innovación.

Two immobilized preparations from Thermomyces lanuginosus lipase (TLL) were compared in the synthesis of butyl butyrate. The commercial Lipozyme TL-IM, and TLL immobilized on styrene–divinylbenzene beads (MCI-TLL) were tested in the esterification reaction using n-hexane as solvent. The variables temperature (30–60 °C), substrate molar ratio (1:1 to 5:1), added water (0–1%), and biocatalyst content (3–40%) were evaluated in terms of initial reaction rate for each biocatalyst. SDS–PAGE analysis revealed that MCI-TLL had an immobilized enzymatic load twice as high as Lipozyme TL-IM, but with an activity 3-fold higher. MCI-TLL presented high initial reaction rates up to 1.0 M butyric acid, while Lipozyme TL-IM showed a decrease in its activity above 0.5 M. Moreover, MCI-TLL allowed a productivity of 14.5 mmol g−1 h−1, while Lipozyme TL-IM 3.2 mmol g−1 h−1, both by mass of biocatalyst., This work was supported by grants from FAPERGS – ARD/2011, from CNPq – Universal/2011, both from Brazil, and CTQ2009-07568 from Spanish Ministerio de Ciencia e Innovación. The authors wish to thank Mr. Ramiro Martínez (Novozymes, Spain) for kindly supplying the enzymes used in this research. We also thank Ministerio de Ciencia e Innovación (Spain) for a fellowship for C. Garcia-Galan, the PhD Program Fellowship-2008 from COLCIENCIAS (Colombia) for a fellowship for O. Barbosa, CNPq (Brazil) for a fellowship to J. L. R. Friedrich, and FAPERGS (Brazil) for a fellowship to A. B. Martins.

[Background] Schistosoma mansoni and Schistosoma japonicum are the most frequent causative agents of human intestinal schistosomiasis. Approximately 200 million people in the world are infected with schistosomes. Diagnosis of schistosomiasis is often difficult. High percentages of low level infections are missed in routine fecal smear analysis and current diagnostic methodologies are inadequate to monitor the progress of parasite control, especially in areas with low transmission. Improved diagnostic methods are urgently needed to evaluate the success of elimination programs. Recently, a magnetic fractionation method for isolation of parasite eggs from feces was described, which uses magnetic microspheres to form parasite egg – magnetic microsphere conjugates. This approach enables screening of larger sample volumes and thus increased diagnostic sensitivity. The mechanism of formation of the conjugates remains unexplained and may either be related to specific surface characteristics of eggs and microspheres or to their magnetic properties., [Methods/Principal Findings] Here, we investigated iron localization in parasite eggs, specifically in the eggshells. We determined the magnetic properties of the eggs, studied the motion of eggs and egg-microsphere conjugates in magnetic fields and determined species specific affinity of parasite eggs to magnetic microspheres. Our study shows that iron is predominantly localized in pores in the eggshell. Parasite eggs showed distinct paramagnetic behaviour but they did not move in a magnetic field. Magnetic microspheres spontaneously bound to parasite eggs without the presence of a magnetic field. S. japonicum eggs had a significantly higher affinity to bind microspheres than S. mansoni eggs., [Conclusions/Significance] Our results suggest that the interaction of magnetic microspheres and parasite eggs is unlikely to be magnetic in origin. Instead, the filamentous surface of the eggshells may be important in facilitating the binding. Modification of microsphere surface properties may therefore be a way to optimize magnetic fractionation of parasite eggs., This work was funded by grants from the Australian Research Council (ARC, DP1093471), the National Health and Medical Research Council of Australia (NHMRC, APP1002898), a University of Western Australia/University of Queensland Bilateral Research Collaboration Award, as well as grants from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, 301979/2010-3) and the Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS, 09/0096-7) in Brazil. Travel to Australia by RLM and RRFC was funded by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, 032/2010) and FAPERGS (12/1137-8 APE) grants awarded to CGT.

This review intends to give a view of the properties and uses of the lipase from Mucor javanicus (MJL) (currently Mucor circinelloides). MJL was described in 1969, its structure is still to be resolved and published. The enzyme is commercialized, but not in any immobilized form; this may have reduced its use by academic groups. This review shows the main features of the enzyme, the immobilization efforts (from mycelium bound enzymes to sophisticated immobilization protocols) and the use of the enzyme in fine chemistry and oil and fats modification. Special interest has been paid to researches where MJL has been compared to other lipases., This work was supported by grants from Fundação de Amparo a Pesquisa do Rio Grande do Sul (FAPERGS–ARD/2011), from CNPq (Brazilian Bureau of Science and Technology) and CTQ2009-07568 and a predoctoral fellowship for Ms. García-Galán from Spanish Ministerio de Ciencia e Innovación, grant No. 1102-489-25428 from COLCIENCIAS and Universidad Industrial de Santander (VIE-UIS Research Program) and the PhD Program Fellowship-2008 for Mr. Barbosa.

14 páginas.-- 8 figuras.-- 80 referencias.-- 4 ficheros adicionales con 4 tablas en http://www.biomedcentral.com/1471-2164/12/409/additional, Background: The species Azospirillum amazonense belongs to a well-known genus of plant growth-promoting bacteria. This bacterium is found in association with several crops of economic importance; however, there is a lack of information on its physiology. In this work, we present a comprehensive analysis of the genomic features of this species. Results: Genes of A. amazonense related to nitrogen/carbon metabolism, energy production, phytohormone production, transport, quorum sensing, antibiotic resistance, chemotaxis/motility and bacteriophytochrome biosynthesis were identified. Noteworthy genes were the nitrogen fixation genes and the nitrilase gene, which could be directly implicated in plant growth promotion, and the carbon fixation genes, which had previously been poorly investigated in this genus. One important finding was that some A. amazonense genes, like the nitrogenase genes and RubisCO genes, were closer phylogenetically to Rhizobiales members than to species of its own order. Conclusion: The species A. amazonense presents a versatile repertoire of genes crucial for its plant-associated lifestyle., This work was supported by grants from the Brazilian National Research Council (CNPq) and the Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS). FHS, RC, LAR and FMS received scholarships from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).