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One of the main distinguishing features of bacteria belonging to the Cellulomonas genus is their ability to secrete multiple polysaccharide degrading enzymes. However, their application in biomass deconstruction still constitutes a challenge. We addressed the optimisation of the xylanolytic activities in extracellular enzymatic extracts of Cellulomonas sp. B6 and Cellulomonas fimi B-402 for their subsequent application in lignocellulosic biomass hydrolysis by culture in several substrates. As demonstrated by secretomic profiling, wheat bran and waste paper resulted to be suitable inducers for the secretion of xylanases of Cellulomonas sp. B6 and C. fimi B-402, respectively. Both strains showed high xylanolytic activity in culture supernatant although Cellulomonas sp. B6 was the most efficient xylanolytic strain. Upscaling from flasks to fermentation in a bench scale bioreactor resulted in equivalent production of extracellular xylanolytic enzymatic extracts and freeze drying was a successful method for concentration and conservation of the extracellular enzymes, retaining 80% activity. Moreover, enzymatic cocktails composed of combined extra and intracellular extracts effectively hydrolysed the hemicellulose fraction of extruded barley straw into xylose and xylooligosaccharides. Instituto de Biotecnología Fil: Ontañon, Ornella. Instituto Nacional de Tecnología Agropecuaria (INTA). UEDD IABIMO. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Ontañon, Ornella. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Bedő, Soma. Budapest University of Technology and Economics. Faculty of Chemical Technology and Biotechnology. Department of Applied Biotechnology and Food Science. Biorefinery Research Group; Hungría Fil: Ghio, Silvina. Instituto Nacional de Tecnología Agropecuaria (INTA). UEDD IABIMO. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Ghio, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Garrido, Mercedes Maria. Instituto Nacional de Tecnología Agropecuaria (INTA). UEDD IABIMO. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Garrido, Mercedes Maria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Topalian, Juliana. Instituto Nacional de Tecnología Agropecuaria (INTA). UEDD IABIMO. Instituto de Agrobiotecnología y Biología Molecular. Fil: Topalian, Juliana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Jahola, Dóra. Budapest University of Technology and Economics. Faculty of Chemical Technology and Biotechnology. Department of Applied Biotechnology and Food Science. Biorefinery Research Group; Hungría Fil: Fehér, Anikó. Budapest University of Technology and Economics. Faculty of Chemical Technology and Biotechnology. Department of Applied Biotechnology and Food Science. Biorefinery Research Group; Hungría Fil: Valacco, Maria Pia. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Centro de Estudios Químicos y Biológicos por Espectrometría de Masa; Argentina Fil: Valacco, Maria Pia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Campos, Eleonora. Instituto Nacional de Tecnología Agropecuaria (INTA). UEDD IABIMO. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Campos, Eleonora. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Fehér, Csaba. Budapest University of Technology and Economics. Faculty of Chemical Technology and Biotechnology. Department of Applied Biotechnology and Food Science. Biorefinery Research Group; Hungría

Woody biomass represents an important source of carbon on earth, and its global recycling is highly dependent on Agaricomycetes fungi. White-rot Basidiomycetes are a very important group in this regard, as they possess a large and diverse enzymatic repertoire for biomass decomposition. Among these enzymes, the recently discovered lytic polysaccharide monooxygenases (LPMOs) have revolutionized biomass processing with their novel oxidative mechanism of action. The strikingly high representation of LPMOs in fungal genomes raises the question of their functional versatility. In this work, we studied an AA9 LPMO from the white-rot basidiomycete Pycnoporus sanguineus, PsAA9A. Successfully produced as a recombinant secreted protein in Pichia pastoris, PsAA9A was found to be a C1-specific LPMO active on cellulosic substrates, generating native and oxidized cello-oligosaccharides in the presence of an external electron donor. PsAA9A boosted cellulolytic activity of glysoside hydrolases from families GH1, GH5, and GH6.This study serves as a starting point towards understanding the functional versatility and biotechnological potential of this enzymatic family, highly represented in wood decay fungi, in Pycnoporus genus. KEY POINTS: • PsAA9A is the first AA9 from P. sanguineus to be characterized. • PsAA9A has activity on cellulose, producing C1-oxidized cello-oligosaccharides. • Boosting activity with GH1, GH5, and GH6 was proven.

This article proposes a historical, multispecies, and ontological approach to human–wildlife conflict (HWC) in the Colombian páramos. Focusing on the páramos surrounding the capital city of Bogotá, we reconstruct the historically changing relationship between cattle-farming campesino communities and the Andean bear, Tremarctos ornatus. Using ethnographic and historical research methods, we conceptualise this relationship as embedded in localised landscapes and multispecies assemblages, in which scientists, conservation practitioners, water infrastructures, public environmental agencies, and cows participate as well. This article demonstrates that insufficient attention to the practices and relationships of historically marginalised humans and non-humans in the management of HWCs contributes to new dynamics of exclusion and friction, and can reduce the effectiveness of conservation programmes. We conclude that opening up conservation to the interests and knowledges of local communities is imperative in moving towards more historically informed, pluralistic and effective conservation strategies.

The gut is home to millions of microscopic organisms, which together are called the gut microbiota. Most of these microorganisms live in peace with the rest of our cells, help us get energy from food, and give us essential nutrients that we cannot make ourselves. The cells lining the gut’s surface keep these microorganisms separated from the rest of the body, like the “Great Wall” of our kingdom. On the inside of this Great Wall, the heroes of the mighty immune system watch over and protect us. But how can the immune system tell the difference between our own cells and the gut microorganisms? Or between helpful microorganisms and disease-causing ones? It is an amazing storey, full of secrets. In this article, we will explain how the gut coaches the immune system to recognise enemies, and how some aspects of modern life might be interfering with this process.

Highlights • Cel8Pa is an extracellular, halotolerant, broad substrate endoglucanase. • Bg1Pa is an intracellular β-glucosidase, with activity on cello oligosaccharides and high resistance to ethanol. • The concerted action of Cel8Pa and Bg1Pa has a synergistic effect on saccharification of β-glucans. • Cel8Pa and Bg1Pa are cold-stable and candidates for SSF ethanol 2 G processes., In the efficient bioconversion of polysaccharides from lignocellulosic biomass, endoglucanases and β-glucosidases are key enzymes for the deconstruction of β-glucans. In this work, we focused on a GH8 endoglucanase (Cel8Pa) and a GH1 β-glucosidase (Bg1Pa) from Paenibacillus xylanivorans A59. Cel8Pa was active on a broad range of substrates, such as β-glucan from barley (24.5 IU/mg), lichenan (17.9 IU/mg), phosphoric acid swollen cellulose (PASC) (9.7 IU/mg), carboxi-methylcellulose (CMC) (7.3 IU/mg), chitosan (1.4 IU/mg) and xylan (0.4 IU/mg). Bg1Pa was active on cellobiose (C2) and cello-oligosaccharides up to C6, releasing glucose as the main product. When both enzymes were used jointly, there was a synergic effect in the conversion rate of polysaccharides to glucose. Cel8Pa and Bg1Pa presented important properties for simultaneous saccharification and fermentation (SSF) processes in second generation bioethanol production, such as tolerance to high concentration of glucose and ethanol.

Xylanases are key enzymes for agricultural biomass saccharification for the production of cellulosic ethanol. Success in enzymatic lignocellulose bioconversion is restricted by enzyme production costs, activity and stability under harsh reaction conditions, and their performance when interacting into cellulolytic cocktails. In this work, we present the heterologous expression and enzymatic characterization of a novel endo-β-1,4 xylanase of glycoside hydrolase family 10 (GH10ps) from the white-rot basidiomycete Pycnoporus sanguineus BAFC 2126. Recombinant expression of GH10ps in Pichia pastoris showed that it is a robust enzyme active at a wide range of pHs and temperatures, and with a half-life of 3 h at 70 °C and a stability higher than 48 h at 60 °C. Recombinant GH10ps was also capable of releasing xylooligosaccharides and xylose from pretreated agricultural waste biomass and also complemented commercial cellulases in lignocellulose bioconversion to fermentable sugars. Fil: Niderhaus, Cecilia. Universidad de Buenos Aires; Argentina Fil: Garrido, Mercedes María. Universidad de Buenos Aires; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Insani, Ester Marina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Campos, Eleonora. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Wirth, Sonia Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina

Glycoside hydrolase family 8 (GH8) includes endoglucanases, lichenases, chitosanases and xylanases, which are essential for polysaccharides breakdown. In this work, we studied a thermally stable GH8 from the cellulose synthase complex of Enterobacter sp. R1, for deconstruction of β-glucans. The biochemical characterization of the recombinant GH8ErCel showed high specificity towards barley β-glucan and lichenan and lower activity on carboxymethylcellulose and swollen cellulose, yielding different length oligosaccharides. By molecular modeling, six conserved subsites for glucose binding and some possible determinants for its lack of xylanase and chitosanase activity were identified. GH8ErCel was active at a broad range of pH and temperature and presented remarkable stability at 60 °C. Additionally, it hydrolyzed β-glucan from oat and wheat brans mainly to tri- and tetraoligosaccharides. Therefore, GH8ErCel may be a good candidate for enzymatic deconstruction of β-glucans at high temperature in food and feed industries, including the production of prebiotics and functional foods.

Little is known about the pathogenesis of cerebral sinovenous thrombosis (CSVT) in the neonate. Although thrombophilia has been described as increasing the risk of CSVT in adults, it remains controversial in pediatric patients, and prospective case–control studies regarding neonatal CSVT are lacking. From 2008 to 2017, all 26 consecutive newborn infants ≥35 weeks of gestation diagnosed with neonatal CSVT, and their mothers, were tested for factor V Leiden (FV) G1691A, FII G20210A, and methylenetetrahydrofolate reductase C677T (MTHFR C677T) mutations. Eighty-five mother–infant pairs were recruited as controls. All infants except 1 with CSVT were suspected due to clinical symptoms, mainly seizures (22/25). Magnetic resonance imaging was performed in 24/26 infants. Heterozygous FV G1691A, FII G20210A, and homozygous MTHFR C677T mutations were present in 1/26, 3/26, and 3/20 infants with CSVT, respectively. FII (odds ratio: 10.96; 95% confidence interval [CI]: 1.09-110.35) and male sex (3.93; 95% CI: 1.43-10.76) were associated with CSVT. When FII G20210A analysis was adjusted for sex, the OR for FII G20210A was 6.70 (95% CI: 0.65-69.22). No differences were found for FV G1691A or homozygous MTHFR mutations between neonates with CSVT and their mothers, compared to controls.

Gender is a crucial determinant of life span, but little is known about gender differences in free radical homeostasis and inflammatory signaling. The aim of the study was to determine gender-related differences concerning oxidative stress and inflammatory signaling of healthy neonates and mothers. Fifty-six mothers with normal gestational course and spontaneous delivery were selected. Blood samples were collected from the mother (at the beginning of delivery and start of expulsive period) and from neonate (from umbilical cord vein and artery). The mothers of girls featured a higher total antioxidant status and lower plasma hydroperoxides than the mother of boys. Regarding the neonates, the girls featured a higher total antioxidant status and lower plasma membrane hydroperoxides in umbilical cord artery together with higher catalase, glutathione peroxidase, and superoxide dismutase activities. Lower levels of interleukin 6, tumor necrosis factor alpha, and prostaglandin E2 were observed in the mothers of girls and higher level of soluble tumor necrosis factor receptor II. In the neonates, lower levels of interleukin 6 and tumor necrosis factor alpha were observed in umbilical artery and higher soluble tumor necrosis factor receptor II in umbilical cord vein and artery of girls. An association between gender, oxidative stress, and inflammation signaling exists, leading to a renewed interest in the neonate’s sex as a potential risk factor to several alterations.

UNLABELLED Hyperammonaemia/mild hepatic encephalopathy (HE) can be simulated by the oral administration of a so-called amino acid challenge (AAC). This study sought to assess the effects of the AAC alone and in combination with either ammonia-lowering [L-ornithine-L-aspartate (LOLA)] or vigilance-enhancing medication (caffeine). Six patients with cirrhosis (5 males; 61.3 ± 9.2 years; 5 Child A, 1 Child B) and six healthy volunteers (5 males; 49.8 ± 10.6 years) were studied between 08:00 and 19:00 on Monday of three consecutive weeks. The following indices were obtained: hourly capillary ammonia, hourly subjective sleepiness, paper & pencil/computerized psychometry and wake electroencephalography (EEG) at 12:00, i.e. at the time of the maximum expected effect of the AAC. RESULTS On average, patients had worse neuropsychological performance and slower EEG than healthy volunteers in all conditions but differences did not reach significance. In healthy volunteers, the post-AAC increase in capillary ammonia levels was contained by both the administration of LOLA and of caffeine (significant differences between 10:00 and 14:00 h). The administration of caffeine also resulted in a reduction in subjective sleepiness and in the amplitude of the EEG on several frontal/temporal-occipital sites (p

Introduction Macrophages play a major role as effector cells of the innate immune system and are vital for intestinal tissue homeostasis. An altered function of intestinal macrophages may contribute to the development and propagation of intestinal inflammation in IBD. However, all data available come from mouse models, human whole tissue or in vitro derived macrophages from blood monocytes. We have isolated intestinal macrophages from patients and healthy subjects and analysed their transcriptome in order to study the intrinsic role that macrophages play in the pathogenesis of IBD. Method Fresh colonic mucosal tissue biopsies from 10 CD patients, 10 UC patients and 10 healthy controls were disaggregated to cell suspensions and sorted using fluorescence-activated cell sorting. RNA from intestinal macrophages, identified as CD163+CD14+CD3- population, was extracted and subjected to RNA sequencing. Differential Gene Expression analysis and pathway analysis were performed between the groups. Results The transcriptomic analysis revealed that the gene expression profile of the intestinal macrophages from IBD is dramatically reprogrammed. Differential Gene Expression analysis revealed 1287 DEGs between macrophages from UC patients and healthy controls; 840 DEGs between macrophages from CD patients and healthy controls and 20 DEGs between macrophages from UC and CD patients (1.5 fold change and FDR Conclusion This is the first study to describe the transcriptome of intestinal macrophages from active lesions of patients with IBD by high throughput RNAseq. We show that the transcriptome of these macrophages is profoundly different from those taken from healthy subjects. These results suggest that macrophages play an important role in the propagation of inflammation and we have identified a number of molecules that should be investigated as potential therapeutic targets. Disclosure of Interest None Declared