Carbon source pulsed feeding to attain high yield and high productivity in poly(3-hydroxybutyrate) (PHB) production from soybean oil using Cupriavidus necator

Poly(3-hydroxybutyrate) (PHB) biosynthesis from soybean oil by Cupriavidus necator was studied using a bench scale bioreactor. The highest cell concentration (83 g l(-1)) was achieved using soybean oil at 40 g l(-1) and a pulse of the same concentration. The PHB content was 81% (w/w), PHB productivity was 2.5 g l(-1) h(-1), and the calculated Y-p/s value was 0.85 g g(-1). Growth limitation and the onset of PHB biosynthesis took place due to exhaustion of P, and probably also Cu, Ca, and Fe.

A quantitative approach to evolution of music and philosophy

The development of new statistical and computational methods is increasingly making it possible to bridge the gap between hard sciences and humanities. In this study, we propose an approach based on a quantitative evaluation of attributes of objects in fields of humanities, from which concepts such as dialectics and opposition are formally defined mathematically. As case studies, we analyzed the temporal evolution of classical music and philosophy by obtaining data for 8 features characterizing the corresponding fields for 7 well-known composers and philosophers, which were treated with multivariate statistics and pattern recognition methods. A bootstrap method was applied to avoid statistical bias caused by the small sample data set, with which hundreds of artificial composers and philosophers were generated, influenced by the 7 names originally chosen. Upon defining indices for opposition, skewness and counter-dialectics, we confirmed the intuitive analysis of historians in that classical music evolved according to a master apprentice tradition, while in philosophy changes were driven by opposition. Though these case studies were meant only to show the possibility of treating phenomena in humanities quantitatively, including a quantitative measure of concepts such as dialectics and opposition, the results are encouraging for further application of the approach presented here to many other areas, since it is entirely generic.

Generation of a Chinese Hamster Ovary Cell Line Producing Recombinant Human Glucocerebrosidase

Impaired activity of the lysosomal enzyme glucocerebrosidase (GCR) results in the inherited metabolic disorder known as Gaucher disease. Current treatment consists of enzyme replacement therapy by administration of exogenous GCR. Although effective, it is exceptionally expensive, and patients worldwide have a limited access to this medicine. In Brazil, the public healthcare system provides the drug free of charge for all Gaucher's patients, which reaches the order of $ 84million per year. However, the production of GCR by public institutions in Brazil would reduce significantly the therapy costs. Here, we describe a robust protocol for the generation of a cell line producing recombinant human GCR. The protein was expressed in CHO-DXB11 (dhfr(-)) cells after stable transfection and gene amplification with methotrexate. As expected, glycosylated GCR was detected by immunoblotting assay both as cell-associated (similar to 64 and 59 kDa) and secreted (63-69 kDa) form. Analysis of subclones allowed the selection of stable CHO cells producing a secreted functional enzyme, with a calculated productivity of 5.14 pg/cell/day for the highest producer. Although being laborious, traditionalmethods of screening high-producing recombinant cellsmay represent a valuable alternative to generate expensive biopharmaceuticals in countries with limited resources.

A light in the shadow: the use of Lucifer Yellow technique to demonstrate nectar reabsorption

Abstract Background Nectar reabsorption is a widely known phenomenon, related to the strategy of resource-recovery and also to maintain the nectar homeostasis at the nectary. The method currently performed to demonstrate nectar being reabsorbed involves the use of radioactive tracers applied to the nectary. Although this method works perfectly, it is complex and requires specific supplies and equipment. Therefore, here we propose an efficient method to obtain a visual demonstration of nectar reabsorption, adapting the use of Lucifer Yellow CH (LYCH), a fluorescent membrane-impermeable dye that can enter the vacuole by endocytosis. Results We applied a LYCH solution to the floral nectary (FN) of Cucurbita pepo L., which is a species known for its ability of nectar reabsorption, and to the extrafloral nectary (EFN) of Passiflora edulis Sims which does not reabsorb the secreted nectar. In all tests performed, we observed that LYCH stained the nectary tissues differentially according to the reabsorption ability of the nectary. The treated FN of C. pepo presented a concentrated fluorescence at the epidermis that decreased at the deeper nectary parenchyma, until reaching the vascular bundles, indicating nectar reabsorption in the flowers of the species. In contrast, treated EFN of P. edulis presented fluorescence only at the cuticle surface, indicating that nectar is not reabsorbed by that particular tissue. Conclusion LYCH is an efficient marker to demonstrate nectar reabsorption.

Quantitative correlation between transcriptional levels of ER chaperone, peroximal protein and FVIII productivity in human Hek-293 cell line

Hek-293 cell line presents good production platform for recombinant therapeutic proteins, however little is known about the components that contribute to the cellular control of recombinant protein production. In this study, we generated a Hek-293 producing recombinant factor VIII (FVIII) and we evaluated the immunoglobulin-binding protein (BiP) and phytanoil-CoA α-hydroxylase (PAHX) expression levels which are known for diminishing FVIII production. Our analyses showed that the recombinant cell population expresses 3.1 ± 1.4 fold of BIP mRNA (P = 0.0054) and 97.8 ± 0.5 fold of PAHX mRNA (P = 0.0016) compared to nontransduced cells. The amount of these proteins was inversely correlated to the secreted FVIII. In conclusion, BIP and PAHX expression are augmented in human cells producing FVIII and they antagonize the amount of therapeutic factor VIII in the cell culture.

The secrets of publishing your article in Journal of Geophysical Research (Oceans)

[EN] The editorial and review processes along the road to publication are described in general terms. The construction of a well-prepared article and the manner in which authors may maximise the chances of success at each stage of the process towards final publication are explored. The most common errors and ways of avoiding them are outlined. Typical problems facing an author writing in English as a second language, including the need for grammatical precision and appropriate style, are discussed. Additionally, the meaning of plagiarism, self-plagiarism and duplicate publication is explored. Critical steps in manuscript preparation and response to reviews are examined. Finally, the relation between writing and reviewing is outlined, and it is shown how becoming a good reviewer helps in becoming a successful author

Semantic Publishing: issues, solutions and new trends in scholarly publishing within the Semantic Web era

This work is concerned with the increasing relationships between two distinct multidisciplinary research fields, Semantic Web technologies and scholarly publishing, that in this context converge into one precise research topic: Semantic Publishing. In the spirit of the original aim of Semantic Publishing, i.e. the improvement of scientific communication by means of semantic technologies, this thesis proposes theories, formalisms and applications for opening up semantic publishing to an effective interaction between scholarly documents (e.g., journal articles) and their related semantic and formal descriptions. In fact, the main aim of this work is to increase the users' comprehension of documents and to allow document enrichment, discovery and linkage to document-related resources and contexts, such as other articles and raw scientific data. In order to achieve these goals, this thesis investigates and proposes solutions for three of the main issues that semantic publishing promises to address, namely: the need of tools for linking document text to a formal representation of its meaning, the lack of complete metadata schemas for describing documents according to the publishing vocabulary, and absence of effective user interfaces for easily acting on semantic publishing models and theories.

Linked Data e bibliometriche: un indice di multidisciplinarieta nel Semantic Publishing

Nel presente lavoro si introduce un nuovo indice per la valutazione dei prodotti della ricerca: l'indice di multidisciplinarieta`. Questa nuova metrica puo` essere un interessante parametro di valutazione: il panorama degli studi multidisciplinari e` vasto ed eterogeneo, ed all'interno di questo sono richieste necessarie competenze trasversali. Le attuali metriche adottate nella valutazione di un accademico, di un journal, o di una conferenza non tengono conto di queste situazioni intermedie, e limitano la loro valutazione dell'impatto al semplice conteggio delle citazioni ricevute. Il risultato di tale valutazione consiste in un valore dell'impatto della ricerca senza una connotazione della direzione e della rilevanza di questa nel contesto delle altre discipline. L'indice di multidisciplinarieta` proposto si integrerebbe allora all'interno dell'attuale panorama delle metriche di valutazione della ricerca, offrendo -accanto ad una quantificazione dell'impatto- una quantificazione della varieta` dei contesti disciplinari nei quali si inserisce.

Semantic publishing: analisi di Linked Open Dataset

Il presente lavoro si occupa di fare una rassegna esaustiva di alcuni Linked Open Dataset nel contesto delle pubblicazioni scientifiche, cercando di inquadrare la loro eterogeneità ed identificando i principali pregi e difetti di ciascuno. Inoltre, descriviamo il nostro prototipo GReAT (Giorgi's Redundant Authors Tool), creato per il corretto riconoscimento e disambiguazione degli autori.

Projected 21st century decrease in marine productivity: a multi-model analysis

Changes in marine net primary productivity (PP) and export of particulate organic carbon (EP) are projected over the 21st century with four global coupled carbon cycle-climate models. These include representations of marine ecosystems and the carbon cycle of different structure and complexity. All four models show a decrease in global mean PP and EP between 2 and 20% by 2100 relative to preindustrial conditions, for the SRES A2 emission scenario. Two different regimes for productivity changes are consistently identified in all models. The first chain of mechanisms is dominant in the low- and mid-latitude ocean and in the North Atlantic: reduced input of macro-nutrients into the euphotic zone related to enhanced stratification, reduced mixed layer depth, and slowed circulation causes a decrease in macro-nutrient concentrations and in PP and EP. The second regime is projected for parts of the Southern Ocean: an alleviation of light and/or temperature limitation leads to an increase in PP and EP as productivity is fueled by a sustained nutrient input. A region of disagreement among the models is the Arctic, where three models project an increase in PP while one model projects a decrease. Projected changes in seasonal and interannual variability are modest in most regions. Regional model skill metrics are proposed to generate multi-model mean fields that show an improved skill in representing observation-based estimates compared to a simple multi-model average. Model results are compared to recent productivity projections with three different algorithms, usually applied to infer net primary production from satellite observations.