Effects of lignin content on the properties of lignocellulose-based biocomposites

The paper is focused on the analysis of the behaviour of biocomposites (biodegradable composites) which are reinforced with different fillers fractions, with varying lignin contents. These materials have been carried-out by extrusion and injection moulding. The matrix, an aromatic copolyester (polybutylene adipate-co-terephthalate), is biodegradable. The lignocellulose fillers are a by-product of an industrial fractionation process of wheat straw. From the raw agro-material and by lignin extractions, various fillers fractions have been obtained by varying the fractionation conditions, both on the liquid media (aqueous or organic) and on the temperature. The fillers lignin contents vary from 30 to 14 wt% with a resultant increase of the cellulose content. We have analysed the impact of the different extraction conditions on the fillers surface and size distribution, and also on the final thermal and mechanical properties of the biocomposites. These materials present significant differences of behaviour which can fulfil some requirements for applications, such as non-food packaging or other short-lived applications (agriculture, sport …) where long-lasting polymers are not entirely adequate.     

Pitfalls in the sample preparation and analysis of N-acylethanolamines

N-acylethanolamines (NAEs) are a group of lipid mediators synthesized in response to a number of physiological and pathological stimuli. Because of the low tissue concentrations of NAEs, analyses often include liquid extraction followed by solid-phase extraction and subsequent quantitation by LC/MS or GC/MS. Reported levels of NAEs vary considerably, however, and often no explanation is given for these discrepancies. Brought on by difficulties encountered during method development, the effects of using four different brands of silica-containing solid phase extraction (SPE) columns and five different brands of chloroform for sample preparation were investigated. Considerable variation in the retention and recoveries of seven NAEs and 2-arachidonoylglycerol existed between the SPE columns. Furthermore, it was found that some chloroforms contained quantifiable amounts of N-palmitoylethanolamine and N-stearoylethanolamine. Finally, it was found that use of one of the chloroforms resulted in a loss of N-oleoylethanolamine from solution due to addition of chlorine to the ω-9 bond. The identity of this reaction product was confirmed by LC-MS/MS and NMR. It is recommended that these aspects of sample preparation and analysis should be thoroughly validated during method development and the relevant information on specific brands used be reported in future communications in order to better estimate the validity of reported quantitative data.     

A Rapid Method for Protein Extraction from Fission Yeast

Researchers working with fission yeast conduct protein extraction widely and frequently, but this includes the handling of glass beads, and hence is laborious and cumbersome, especially when dealing with a large number of samples. Here we describe a rapid and reliable method for preparing protein extract from fission yeast, one which is applicable to routine western blotting.     

Human Health Risk Assessment of Soils Contaminated with Metal(loid)s by Using DGT Uptake: A Case Study of a Former Korean Metal Refinery Site

The human health risk of soils contaminated with As, Pb, Cu, and Zn was evaluated based on pseudo-total concentrations of metal(loid)s, the physiologically based extraction test (PBET), and diffusive gradients in thin films (DGT). Non-carcinogenic (NCR) and carcinogenic (CR) risks exceeded the U.S. Environmental Protection Agency criteria under both the residential and non-residential scenarios. Human bioavailable concentrations (PBET) were much lower than pseudo-total concentrations. The Hazardous Index of NCR (HI (NCR)) for the PBET in the studied soils was 67% and 94% less than that for pseudo-total concentration, respectively, under the non-residential and residential scenarios. Similarly, CR for the PBET was also 65% and 93% less for the two soils. The concentration of metal(loid)s accumulated in the DGT resin was highly correlated with the PBET-extractable concentration (R² > 0.649). Therefore, for both the CR and HI (NCR), the DGT-calculated risk was linearly related to the PBET-calculated risk for the studied soils under both scenarios. The results suggest that DGT uptake and PBET-extracted concentrations are good surrogates for risk estimation and that both J1 and J2 soils require remediation before their use for residential or non-residential purposes.     

An Optimized Proteomics Approach Reveals Novel Alternative Proteins in Mouse Liver Development

Alternative ORFs (AltORFs) are unannotated sequences in genome that encode novel peptides or proteins named alternative proteins (AltProts). Although ribosome profiling and bioinformatics predict a large number of AltProts, mass spectrometry as the only direct way of identification is hampered by the short lengths and relative low abundance of AltProts. There is an urgent need for improvement of mass spectrometry methodologies for AltProt identification. Here, we report an approach based on size-exclusion chromatography for simultaneous enrichment and fractionation of AltProts from complex proteome. This method greatly simplifies the variance of AltProts discovery by enriching small proteins smaller than 40 kDa. In a systematic comparison between 10 methods, the approach we reported enabled the discovery of more AltProts with overall higher intensities, with less cost of time and effort compared to other workflows. We applied this approach to identify 89 novel AltProts from mouse liver, 39 of which were differentially expressed between embryonic and adult mice. During embryonic development, the upregulated AltProts were mainly involved in biological pathways on RNA splicing and processing, whereas the AltProts involved in metabolisms were more active in adult livers. Our study not only provides an effective approach for identifying AltProts but also novel AltProts that are potentially important in developmental biology.     

Extraction and Antioxidant Activities of Magnolia kwangsiensis Figlar & Noot. Leaf Polyphenols

Microwave‐assisted extraction was employed to extract polyphenols from the leaf of Magnolia kwangsiensis Figlar & Noot . The yield of polyphenols was 2.44±0.02 % under the optimal conditions of RSM: acetone concentration of 70 %, ratio of solvent to material of 21 mL?g^?1 and extraction time of 16 min. The antioxidant activities were evaluated in terms of total antioxidant ability, reducing power, DPPH ^? and ^? OH scavenging activity. Results showed the polyphenols presented potential antioxidant activities, especially the stronger scavenging activity on ^? OH. In term of ^? OH scavenging activity, the IC₅₀ value of NKA‐9 purification was 0.335 mg mL^?1, equivalent to 35.23 % of V_C. The IC₅₀ values of crude extract and ethyl acetate extract were 0.580 and 0.828 mg mL^?1, equivalent to 60.99 % and 87.07 % of V_C. Results indicated that M. kwangsiensis leaf polyphenols present potential antioxidant activities that make it beneficial for human health by preventing or reducing oxidative damage.     

Fractionation of Heavy Metals in Mine Tailings Amended with Composted Manure

This study evaluated the effect of composted cow manure (CCM) on the chemical fractionation and retention degree of heavy metals (HMs) in mine tailings from Zimapán, México. In a greenhouse experiment, mine tailings from three deposits were incubated for 3 months; experimental units were placed in a PVC container, where increasing doses of CCM were applied. HM pseudo-total concentrations, HM extractions with ethylenediaminetetraacetic acid (EDTA, 0.05 M), and a sequential chemical extraction (SCE) were carried out. The HM concentrations were determined by atomic absorption spectrophotometry. The pseudo-total concentrations of Pb, Cu, Cd, and Ni found were up to 1506, 206, 27, and 23 mg kg^?1, respectively; extractable Pb was up to 42%; 21% for Cu; 51% for Cd; and 16% of Ni of the pseudo-total concentrations of each metal. Treatment with 12% of CCM in mine tailing decreased EDTA-extractable HM concentrations, while the SCE revealed a decrease in exchangeable fraction and an increase in the organic fraction of HM. A positive correlation between CCM application and organic fractions of HMs was found, although the highest increasements were recorded in the organic fraction.     

火龙果茎基因组DNA提取方法改良

火龙果(Hylocereus undulatus)是近年发展起来的一种新兴热带水果, 其茎富含多糖、多酚及其它次生代谢物, 黏性极大, 很难从中提取高质量的DNA。特别是一年生以上的老茎, 目前尚未有较好的DNA提取方法。为了解决这一难题, 该研究对CTAB+Tris-HCl洗涤法进行了3种方式的改良。结果表明, “改进三”方法可不受取样时期和取样部位的限制, 从一年生以上火龙果茎中提取的DNA质量最好且不含黏性物质, 可用于酶切与分子标记等生化和分子生物学实验。该研究探索了一条较为理想的火龙果茎DNA提取方法, 值得推广应用。     

十字花科植物种子低分子RNA提取方法比较

非编码RNA不翻译成蛋白质,它们通过转录、转录后及翻译水平调控靶基因表达,在植物生长发育及逆境胁迫中发挥功能。目前,大量种子萌发期特异表达的非编码RNA (Non-coding RNA)已被发现,高效提取种子低分子RNA是对其进行研究的关键。本研究将介绍一种改良SDS RNA提取方法,并与Trizol、CTAB法、RNA提取试剂盒进行比较。结果表明:这种方法可以高效提取用于Northern blotting、RT-PCR等分子生物学分析的十字花科植物种子低分子RNA。改良SDS RNA提取方法为种子非编码RNA研究、种子萌发生理及分子育种研究提供了帮助。     

Have the cake and eat it: Optimizing nondestructive DNA metabarcoding of macroinvertebrate samples for freshwater biomonitoring

DNA metabarcoding can contribute to improving cost‐effectiveness and accuracy of biological assessments of aquatic ecosystems, but significant optimization and standardization efforts are still required to mainstream its application into biomonitoring programmes. In assessments based on freshwater macroinvertebrates, a key challenge is that DNA is often extracted from cleaned, sorted and homogenized bulk samples, which is time‐consuming and may be incompatible with sample preservation requirements of regulatory agencies. Here, we optimize and evaluate metabarcoding procedures based on DNA recovered from 96% ethanol used to preserve field samples and thus including potential PCR inhibitors and nontarget organisms. We sampled macroinvertebrates at five sites and subsampled the preservative ethanol at 1 to 14 days thereafter. DNA was extracted using column‐based enzymatic (TISSUE) or mechanic (SOIL) protocols, or with a new magnetic‐based enzymatic protocol (BEAD), and a 313‐bp COI fragment was amplified. Metabarcoding detected at least 200 macroinvertebrate taxa, including most taxa detected through morphology and for which there was a reference barcode. Better results were obtained with BEAD than SOIL or TISSUE, and with subsamples taken 7–14 than 1–7 days after sampling, in terms of DNA concentration and integrity, taxa diversity and matching between metabarcoding and morphology. Most variation in community composition was explained by differences among sites, with small but significant contributions of subsampling day and extraction method, and negligible contributions of extraction and PCR replication. Our methods enhance reliability of preservative ethanol as a potential source of DNA for macroinvertebrate metabarcoding, with a strong potential application in freshwater biomonitoring.