Implications of cellobiohydrolase glycosylation for use in biomass conversion

The cellulase producing ascomycete, Trichoderma reesei (Hypocrea jecorina), is known to secrete a range of enzymes important for ethanol production from lignocellulosic biomass. It is also widely used for the commercial scale production of industrial enzymes because of its ability to produce high titers of heterologous proteins. During the secretion process, a number of post-translational events can occur, however, that impact protein function and stability. Another ascomycete, Aspergillus niger var. awamori, is also known to produce large quantities of heterologous proteins for industry. In this study, T. reesei Cel7A, a cellobiohydrolase, was expressed in A. niger var. awamori and subjected to detailed biophysical characterization. The purified recombinant enzyme contains six times the amount of N-linked glycan than the enzyme purified from a commercial T. reesei enzyme preparation. The activities of the two enzyme forms were compared using bacterial (microcrystalline) and phosphoric acid swollen (amorphous) cellulose as substrates. This comparison suggested that the increased level of N-glycosylation of the recombinant Cel7A (rCel7A) resulted in reduced activity and increased non-productive binding on cellulose. When treated with the N-glycosidase PNGaseF, the molecular weight of the recombinant enzyme approached that of the commercial enzyme and the activity on cellulose was improved

COBRA-SFS: A thermal-hydraulic analysis code for spent fuel storage and transportation casks

COBRA-SFS is a general thermal-hydraulic analysis computer code for prediction of material temperatures and fluid conditions in a wide variety of systems. The code has been validated for analysis of spent fuel storage systems, as part of the Commercial Spent Fuel Management Program of the US Department of Energy. The code solves finite volume equations representing the conservation equations for mass, moment, and energy for an incompressible single-phase heat transfer fluid. The fluid solution is coupled to a finite volume solution of the conduction equation in the solid structure of the system. This document presents a complete description of Cycle 2 of COBRA-SFS, and consists of three main parts. Part 1 describes the conservation equations, constitutive models, and solution methods used in the code. Part 2 presents the User Manual, with guidance on code applications, and complete input instructions. This part also includes a detailed description of the auxiliary code RADGEN, used to generate grey body view factors required as input for radiative heat transfer modeling in the code. Part 3 describes the code structure, platform dependent coding, and program hierarchy. Installation instructions are also given for the various platform versions of the code that are available

Floral uniformity through evolutionary time in a species-rich tree lineage

Changes in floral morphology are expected across evolutionary time and are often promoted as important drivers in angiosperm diversification. Such a statement, however, is in contrast to empirical observations of species-rich lineages that show apparent conservative floral morphologies even under strong selective pressure to change from their environments. Here, we provide quantitative evidence for prolific speciation despite uniform floral morphology in a tropical species-rich tree lineage. We analyse floral disparity in the environmental and phylogenetic context of Myrcia (Myrtaceae), one of the most diverse and abundant tree genera in Neotropical biomes. Variation in floral morphology among Myrcia clades is exceptionally low, even among distantly related species. Discrete floral specialisations do occur, but these are few, present low phylogenetic signal, have no strong correlation with abiotic factors, and do not affect overall macroevolutionary dynamics in the lineage. Results show that floral form and function may be conserved over large evolutionary time scales even in environments full of opportunities for ecological interactions and niche specialisation. Species accumulation in diverse lineages with uniform flowers apparently does not result from shifts in pollination strategies, but from speciation mechanisms that involve other, nonfloral plant traits

Effects of spoilage on nitrogen and carbon stable isotopes signatures of the clam Ruditapes decussatus

Fish and seafood products are highly susceptible to post-mortem spoilage due to autolytic reactions at start, then microbiological activity and eventually oxidative reactions. Chemical and microbiological parameters are usually used to assess quality and make decisions for protecting public health, but they lack precision in defining which spoilage pathway is occurring at each moment. The objective of this work was to assess the effects of spoilage reactions on nitrogen and carbon stable isotopes in the grooved carpet shell clam, Ruditapes decussatus, and compare them to biochemical indicators of seafood deterioration, in order to better understand the relations between the different spoilage pathways during commercial storage conditions. Clams were kept in a refrigerator at 5 ºC, to simulate normal commercial storage conditions, and sampled in the beginning of the experiment, and after eight, ten and twelve days. Moisture, condition index, percentage edibility, total volatile basic nitrogen (TVB-N), pH, nitrogen and carbon percentages and stable isotopes were determined for each sampling moment. Stable isotope analyses were performed using a Costech Elemental Analyzer (ECS 4010) coupled to a ThermoFinnigan Delta V Advantage. Stable isotopes analysis, especially for nitrogen, proved to be a good tool for the study of clam deterioration. Nitrogen stable isotopes results showed a relation with other spoilage indicators, such as pH and TVB-N, and allowed identifying spoilage specific pathways, such as amino acids decarboxylation and production of volatile nitrogen compounds.info:eu-repo/semantics/publishedVersio

NEOTROPICAL CARNIVORES: a data set on carnivore distribution in the Neotropics

Incluye contenido parcial de los autoresAbstract.Mammalian carnivores are considered a key group in maintaining ecologicalhealth and can indicate potential ecological integrity in landscapes where they occur. Carni-vores also hold high conservation value and their habitat requirements can guide managementand conservation plans. The order Carnivora has 84 species from 8 families in the Neotropicalregion: Canidae; Felidae; Mephitidae; Mustelidae; Otariidae; Phocidae; Procyonidae; andUrsidae. Herein, we include published and unpublished data on native terrestrial Neotropicalcarnivores (Canidae; Felidae; Mephitidae; Mustelidae; Procyonidae; and Ursidae). NEOTRO-PICAL CARNIVORES is a publicly available data set that includes 99,605 data entries from35,511 unique georeferenced coordinates. Detection/non-detection and quantitative data wereobtained from 1818 to 2018 by researchers, governmental agencies, non-governmental organi-zations, and private consultants. Data were collected using several methods including cameratrapping, museum collections, roadkill, line transect, and opportunistic records. Literature(peer-reviewed and grey literature) from Portuguese, Spanish and English were incorporated inthis compilation. Most of the data set consists of detection data entries (n=79,343; 79.7%) butalso includes non-detection data (n=20,262; 20.3%). Of those, 43.3% also include count data(n=43,151). The information available in NEOTROPICAL CARNIVORES will contribute tomacroecological, ecological, and conservation questions in multiple spatio-temporal perspec-tives. As carnivores play key roles in trophic interactions, a better understanding of their distri-bution and habitat requirements are essential to establish conservation management plans andsafeguard the future ecological health of Neotropical ecosystems. Our data paper, combinedwith other large-scale data sets, has great potential to clarify species distribution and relatedecological processes within the Neotropics. There are no copyright restrictions and no restric-tion for using data from this data paper, as long as the data paper is cited as the source of theinformation used. We also request that users inform us of how they intend to use the data

Fossil Carder Bee's nest from the Hominin locality of Taung, South Africa

The Buxton-Norlim Limeworks southwest of Taung, South Africa, is renowned for the discovery of the first Australopithecus africanus fossil, the ‘Taung Child’. The hominin was recovered from a distinctive pink calcrete that contains an abundance of invertebrate ichnofauna belonging to the Coprinisphaera ichnofacies. Here we describe the first fossil bee’s nest, attributed to the ichnogenus Celliforma, from the Plio-Pleistocene of Africa. Petrographic examination of a cell lining revealed the preservation of an intricate organic matrix lined with the calcitic casts of numerous plant trichomes–a nesting behaviour unique to the modern-day carder bees (Anthidiini). The presence of Celliforma considered alongside several other recorded ichnofossils can be indicative of a dry, savannah environment, in agreement with recent work on the palaeoenvironment of Plio-Pleistocene southern Africa. Moreover, the occurrence of ground-nesting bees provides further evidence that the pink calcrete deposits are of pedogenic origin, rather than speleogenic origin as has previously been assumed. This study demonstrates the potential value of insect trace fossils as palaeoenvironmental indicators

Acute gamma-secretase inhibition of nonhuman primate CNS shifts amyloid precursor protein (APP) metabolism from amyloid-beta production to alternative APP fragments without amyloid-beta rebound

The accumulation of amyloid beta (Aβ) in Alzheimer’s disease is caused by an imbalance of production and clearance, which leads to increased soluble Aβ species and extracellular plaque formation in the brain. Multiple Aβ-lowering therapies are currently in development: an important goal is to characterize the molecular mechanisms of action and effects on physiological processing of Aβ, as well as other amyloid precursor protein (APP) metabolites, in models which approximate human Aβ physiology. To this end, we report the translation of the human in vivo stable-isotope-labeling kinetics (SILK) method to a rhesus monkey cisterna magna ported (CMP) nonhuman primate model, and use the model to test the mechanisms of action of a γ-secretase inhibitor (GSI). A major concern of inhibiting the enzymes which produce Aβ (β- and γ-secretase) is that precursors of Aβ may accumulate and cause a rapid increase in Aβ production when enzyme inhibition discontinues. In this study, the GSI MK-0752 was administered to conscious CMP rhesus monkeys in conjunction with in vivo stable isotope labeling, and dose-dependently reduced newly generated CNS Aβ. In contrast to systemic Aβ metabolism, CNS Aβ production was not increased after the GSI was cleared. These results indicate that most of the CNS APP was metabolized to products other than Aβ, including C-terminal truncated forms of Aβ: 1–14, 1–15 and 1–16; this demonstrates an alternative degradation pathway for CNS amyloid precursor protein during γ-secretase inhibition

Ligand binding site superposition and comparison based on Atomic Property Fields: identification of distant homologues, convergent evolution and PDB-wide clustering of binding sites

A new binding site comparison algorithm using optimal superposition of the continuous pharmacophoric property distributions is reported. The method demonstrates high sensitivity in discovering both, distantly homologous and convergent binding sites. Good quality of superposition is also observed on multiple examples. Using the new approach, a measure of site similarity is derived and applied to clustering of ligand binding pockets in PDB

The Antiquity and Evolutionary History of Social Behavior in Bees

A long-standing controversy in bee social evolution concerns whether highly eusocial behavior has evolved once or twice within the corbiculate Apidae. Corbiculate bees include the highly eusocial honey bees and stingless bees, the primitively eusocial bumble bees, and the predominantly solitary or communal orchid bees. Here we use a model-based approach to reconstruct the evolutionary history of eusociality and date the antiquity of eusocial behavior in apid bees, using a recent molecular phylogeny of the Apidae. We conclude that eusociality evolved once in the common ancestor of the corbiculate Apidae, advanced eusociality evolved independently in the honey and stingless bees, and that eusociality was lost in the orchid bees. Fossil-calibrated divergence time estimates reveal that eusociality first evolved at least 87 Mya (78 to 95 Mya) in the corbiculates, much earlier than in other groups of bees with less complex social behavior. These results provide a robust new evolutionary framework for studies of the organization and genetic basis of social behavior in honey bees and their relatives