<Emphasis Type="Italic">Tequilana weber</Emphasis> Agave Bagasse Enzymatic Hydrolysis for the Production of Fermentable Sugars: Oxidative-Alkaline Pretreatment and Kinetic Modeling

As a way to mitigate the harmful effects of fossil fuel utilization, the use of second-generation ethanol has been proposed. However, the microorganisms responsible for its production are not able to degrade structural polysaccharides, so their hydrolysis is necessary. Previously to this work, a factorial experimental design was carried out to investigate the relation between the NaOH and H₂O₂ concentrations with the yield of carbohydrates, and then this variable was optimized by using a response surface method. A study of the hydrolysis process was performed using enzymes to establish a process that maximizes the depolymerization of Agave tequilana fibers after an alkali-oxidative pretreatment with optimal reagents concentrations, this pretreatment was selected because it can remove almost the total content of lignin and destroys efficiently the crystallinity of cellulose fibers with a lower sugar losses and no production of toxic compounds. An orthogonal array using the novel enzymes Cellic CTec 3 and Cellic HTec 3 was performed to determinate the optimal combination of them, which has resulted in a concentration of 165.67 g/L at the supernatant with 82.21 % conversion and a yield of 352.18 g reducing sugars per kilogram of lignocellulosic material in dry basis. These results are 29 % better in comparison with the previous generation of enzymes with a reduction in the enzymatic charge of 82 %.     

Human Pluripotent Stem Cells and Derived Neuroprogenitors Display Differential Degrees of Susceptibility to BH3 Mimetics ABT-263, WEHI-539 and ABT-199

Human embryonic stem cells (hESCs) are hypersensitive to genotoxic stress and display lower survival ability relative to their differentiated progeny. Herein, we attempted to investigate the source of this difference by comparing the DNA damage responses triggered by the topoisomerase I inhibitor camptothecin, in hESCs, human induced pluripotent stem cells (hiPSCs) and hESCs-derived neuroprogenitors (NP). We observed that upon camptothecin exposure pluripotent stem cells underwent apoptosis more swiftly and at a higher rate than differentiated cells. However, the cellular response encompassing ataxia-telangiectasia mutated kinase activation and p53 phosphorylation both on serine 15 as well as on serine 46 resulted very similar among the aforementioned cell types. Importantly, we observed that hESCs and hiPSCs express lower levels of the anti-apoptotic protein Bcl-2 than NP. To assess whether Bcl-2 abundance could account for this differential response we treated cells with ABT-263, WEHI-539 and ABT-199, small molecules that preferentially target the BH3-binding pocket of Bcl-xL and/or Bcl-2 and reduce their ability to sequester pro-apoptotic factors. We found that in the absence of stress stimuli, NP exhibited a higher sensitivity to ABT- 263 and WEHI-539 than hESCs and hiPSCs. Conversely, all tested cell types appeared to be highly resistant to the Bcl-2 specific inhibitor, ABT-199. However, in all cases we determined that ABT-263 or WEHI-539 treatment exacerbated camptothecin-induced apoptosis. Importantly, similar responses were observed after siRNA-mediated down-regulation of Bcl-xL or Bcl-2. Taken together, our results suggest that Bcl-xL contrary to Bcl-2 contributes to ensure cell survival and also functions as a primary suppressor of DNA double-strand brake induced apoptosis both in pluripotent and derived NP cells. The emerging knowledge of the relative dependence of pluripotent and progenitor cells on Bcl-2 and Bcl-xL activities may help to predict cellular responses and potentially manipulate these cells for therapeutic purposes in the near future.     

Hormone-induced differentiation of agranular endoplasmic reticulum in the interstitial cells of the mouse testis

Summary The administration of chorionic gonadotrophin to prepuberal mice results in precocious maturation of the testicular interstitial cells. The cytoplasm of the nine-day-old cells is characterized by abundant lipid droplets, large numbers of glycogen particles and mitochondria. By contrast, the membranous organelles are poorly developed.Human chorionic gonadotrophin brings about mobilization of lipid droplets and glycogen particles, and differentiation of large areas of agranular endoplasmic reticulum.The present observations are in agreement with the reports that human chorionic gonadotrophin increases the secretion of testosterone and that the agranular endoplasmic reticulum is the site of storage of steroid and of the enzymes involved in the biosynthesis of androgens.     

Predicted distribution and movement of Glossina palpalis palpalis (Diptera: Glossinidae) in the wet and dry seasons in the Kogo trypanosomiasis focus (Equatorial Guinea).

The aim of this study was to predict the distribution and movement of populations of the tsetse fly, Glossina palpalis palpalis (Diptera: Glossinidae), in the wet and dry seasons and to analyze the impact of the use of mono-pyramidal traps on fly populations in the Kogo focus in 2004 and 2005. Three Glossina species are present in Kogo: Glossina palpalis palpalis, major HAT vector in West-Central Africa, Glossina caliginea, and Glossina tabaniformis. The apparent density (AD) of G. p. palpalis clearly fell from 1.23 tsetse/trap/day in July 2004 to 0.27 in December 2005. A significant reduction in the mean AD for this species was noted between seasons and years. The diversity of Glossina species was relatively low at all the sampling points; G. p. palpalis clearly predominated over the other species and significantly dropped as a consequence of control activities. The predictive models generated for the seasonal AD showed notable differences not only in the density but in the distribution of the G. p. palpalis population between the rainy and dry season. The mono-pyramidal traps have proven to be an effective instrument for reducing the density of the tsetse fly populations, although given that the Kogo trypanosomiasis focus extends from the southern Equatorial Guinea to northern Gabon, interventions need to be planned on a larger scale, involving both countries, to guarantee the long-term success of control.     

Capsaicin hydrolysis by Candida antarctica lipase

Capsaicin was hydrolysed by lipase B from Candida antarctica into vanillylamine and 8-methyl-6-trans-nonenoic acid. Conversions of 70% were obtained after 72 h at 70 °C in water but decreased to only 15% when capsaicin was solubilized in 15% (v/v) ethanol/water after 72 h at 45 °C. No activity occurred in chloroform/water mixtures. According to our knowledge, this is the first report concerning amide hydrolysis by a lipase.     

Environmental assessment of multi-functional building elements constructed with digital fabrication techniques

Purpose

Digital fabrication is revolutionizing architecture, enabling the construction of complex and multi-functional building elements. Multi-functionality is often achieved through material reduction strategies such as functional or material hybridization. However, these design strategies may increase environmental impacts over the life cycle. The integration of functions may hinder the maintenance and shorten the service life. Moreover, once a building element has reached the end of life, hybrid materials may influence negatively its recycling capacity. Consequently, the aim of this paper is to analyze the influence of multi-functionality in the environmental performance of two digitally fabricated architectural elements: The Sequential Roof and Concrete-Sandstone Composite Slab and to compare them with existing standard elements.

Methods

A method based on the life-cycle assessment (LCA) framework is applied for the evaluation of the environmental implications of multi-functionality in digital fabrication. The evaluation consists of the comparison of embodied impacts between a multi-functional building element constructed with digital fabrication techniques and a conventional one, both with the same building functions. Specifically, the method considers the lifetime uncertainty caused by multi-functionality by considering two alternative service life scenarios during the evaluation of the digitally fabricated building element. The study is extended with a sensitivity analysis to evaluate the additional environmental implications during end-of-life processing derived from the use of hybrid materials to achieve multi-functionality in architecture.

Results and discussion

The evaluation of two case studies of digitally fabricated architecture indicates that their environmental impacts are very sensitive to the duration of their service life. Considering production and life span phases, multi-functional building elements should have a minimum service life of 30 years to bring environmental benefits over conventional construction. Furthermore, the case study of Concrete-Sandstone Composite Slab shows that using hybrid materials to achieve multi-functionality carries important environmental consequences at the end of life, such as the emission of air pollutants during recycling.

Conclusions

The results from the case studies allow the identification of key environmental criteria to consider during the design of digitally fabricated building elements. Multi-functionality provides material efficiency during production, but design adaptability must be a priority to avoid a decrease in their environmental performance. Moreover, the high environmental impacts caused by end-of-life processing should be compensated during design.

     

Targeted profiling of circulating and hepatic bile acids in human, mouse, and rat using a UPLC-MRM-MS-validated method

Bile acids (BAs) are a group of chemically related steroids recognized as regulatory molecules whose profiles can change in different physio-pathological situations. We have developed a sensitive, fast, and reproducible ultraperformance liquid chromatography/multiple reaction monitoring/mass spectrometry method to determine the tissue and sera BA profiles in different species (human, rat, and mouse) by quantifying 31 major and minor BA species in a single 21-min run. The method has been validated according to FDA guidelines, and it generally provides good results in terms of intra- and interday precision (less than 8.6% and 16.0%, respectively), accuracy (relative error measurement between -11.9% and 8.6%), and linearity (R(2) > 0.996 and dynamic ranges between two and four orders of magnitude), with limits of quantification between 2.5 and 20 nM. The new analytical approach was applied to determine BA concentrations in human, rat, and mouse serum and in liver tissue. Our comparative study confirmed and extended previous reports, showing marked interspecies differences in circulating and hepatic BA composition. The targeted analysis revealed the presence of unexpected minoritary BAs, such as tauro-alpha-Muricholic acid in human serum, thus allowing us to obtain a thorough profiling of human samples. Its great sensitivity, low sample requirements (25 μl of serum, 5 mg of tissue), and comprehensive capacity to profile a considerable number of BAs make the present method a good choice to study BA metabolism in physiological and pathological situations, particularly in toxicological studies.