MV-mimicking micelles loaded with PEG-serine-ACP nanoparticles to achieve biomimetic intra/extra fibrillar mineralization of collagen in vitro

Since their discovery, matrix vesicles (MVs) containing minerals have received considerable attention for their role in the mineralization of bone, dentin and calcified cartilage. Additionally, MVs' association with collagen fibrils, which serve as the scaffold for calcification in the organic matrix, has been repeatedly highlighted. The primary purpose of the present study was to establish a MVs–mimicking model (PEG-S-ACP/micelle) in vitro for studying the exact mechanism of MVs-mediated extra/intra fibrillar mineralization of collagen in vivo. In this study, high-concentration serine was used to stabilize the amorphous calcium phosphate (S-ACP), which was subsequently mixed with polyethylene glycol (PEG) to form PEG-S-ACP nanoparticles. The nanoparticles were loaded in the polysorbate 80 micelle through a micelle self-assembly process in an aqueous environment. This MVs–mimicking model is referred to as the PEG-S-ACP/micelle model. By adjusting the pH and surface tension of the PEG-S-ACP/micelle, two forms of minerals (crystalline mineral nodules and ACP nanoparticles) were released to achieve the extrafibrillar and intrafibrillar mineralization, respectively. This in vitro mineralization process reproduced the mineral nodules mediating in vivo extrafibrillar mineralization and provided key insights into a possible mechanism of biomineralization by which in vivo intrafibrillar mineralization could be induced by ACP nanoparticles released from MVs. Also, the PEG-S-ACP/micelle model provides a promising methodology to prepare mineralized collagen scaffolds for repairing bone defects in bone tissue engineering.     

“绿色革命”新进展:赤霉素与氮营养双重调控的表观修饰助力水稻高产高效育种

以半矮秆育种为代表的“绿色革命”极大地提高了作物产量,但也带来氮营养利用效率降低的严重问题。“绿色革命”主要基于调控赤霉素的代谢和信号转导而实现。前期的研究发现,赤霉素信号转导关键因子DELLA蛋白通过调控GRF4而负调控氮素的吸收利用,为半矮秆品系氮利用效率低的问题提供了解决方案。最近的一项研究进一步揭示了GA信号途径与氮响应交叉互作的新机制。该研究发现水稻(Oryza sativa)NGR5是氮素调控分蘖数目的一个关键基因,其表达受氮诱导。通过招募PRC2,NGR5对D14和OsSPL14等分蘖抑制基因所在位点进行H3K27me3甲基化修饰,从而抑制其表达。而在半矮秆背景下超表达NGR5可以提高低氮水平下的水稻产量。NGR5同时也被发现为赤霉素受体GID1的一个新靶标,受到其负调控。该研究发现了调控赤霉素信号通路的新机制,并对高产高效的新一代“绿色革命”育种实践具有重要启示。     

Functional analysis of the C-terminal propeptide of keratinase from Bacillus licheniformis BBE11-1 and its effect on the production of keratinase in Bacillus subtilis

The keratinase from Bacillus licheniformis BBE11-1 is a serine protease and expressed as a pre-pro-precursor. To produce a mature and active keratinase, the propeptide must be cleaved on the C-terminal via cis or trans. In this study, to enhance the production of keratinase in Bacillus subtilis, single amino acid substitutions, single residue deletions and linkers were introduced at the C-terminus of the propeptide. The results showed that optimizing the residue of cleavage site of propeptide will affect the cleavage efficiency of propeptide, and the mature enzyme yield of Leu(P1)Ala mutant increases 50% compared with the wild-type. In addition, inserting linkers and deleting individual residues at the C-terminal of the propeptide decreases the mature keratinase production. Our results indicated that the primary structure of the C-terminus of propeptide is crucial for the mature keratinase production. Propeptide engineering at C-terminus may be an effective approach to increase the yield of keratinase.     

Gene expression changes associated with cytotoxicity identified using cDNA arrays

In order to investigate gene expression changes associated with cytotoxicity, we used cDNA arrays to monitor the expression of over 5,000 genes in response to toxic stress in the HepG2 liver cell line. Cells were treated with cytotoxic doses of acetaminophen, caffeine or thioacetamide for nine time points ranging from 1 to 24 h. Samples of mRNA from each time point were used to prepare radiolabeled cDNA, which was hybridized to nylon-membrane-based cDNA arrays. High-stringency washes were applied to reduce cross-hybridization. Analysis of spot intensities revealed that each compound led to approximately 150-250 gene expression changes that were sustained over at least three adjacent time points. The affected genes could be classified into clusters based on their temporal patterns of differential expression. A common set of 44 genes showed similar expression changes in response to all three compounds. Of these changes, 90% could be confirmed by quantitative RT-PCR analysis. The results indicate that detailed array-based time-course studies, coupled with a sensitive and highly specific confirmation assay, provide a powerful means of identifying cytotoxicity-associated gene expression changes. Electronic Publication     

The Effect of Leaf Age on Gas Exchange and Malate Accumulation in C3-CAM Plant Marrubium Frivaldszkyanum (Lamiaceae)

For the first time the expression of C₃ and CAM in the leaves of different age of Marrubium frivaldszkyanum Boiss, is reported. With increasing leaf age a typical C₃ photosynthesis pattern and high transpiration rate were found. In older leaves a shift to CAM occurred and the 24-h transpiration water loss decreased. A correlation was established between leaf area and accumulation of malate. Water loss at early stages of leaf expansion may be connected with the shift to CAM and the water economy of the whole plant.     

Food utilization by insects: Interpretation of observed differences between dry weight and energy efficiencies

Over 80% of the values of approximate digestibility (AD), efficiency of conversion of assimilated food to biomass (ECD) and efficiency of conversion of ingested food (ECI) calculated using energy terms are greater than the corresponding dry weight (DW) values, based on data for over 65 species (38 studies; number of comparative values: AD=139, ECD=128 and ECI=169). Largest positive differences (energy > DW values) are 30 (AD, ECD) and 24 (ECI) percentage points and largest negative differences (energy < DW values) are 9 (AD), 11 (ECD) and 8 (ECI) percentage points. These differences generally are least for ECI (71% of the differences fall between 0 and +5 percentage points), and AD (68%), followed by ECD (only 47% fall between 0 and +5), and they may vary with temperature, food and other factors. The differences tend to increase (esp. for ECD and ECI) when comparing later with earlier instars. Energy > DW efficiency values are commonly expected for AD because of the generally greater energy content of food than feces, and for ECD and ECI because of the generally greater energy content of insect biomass than ingested and assimilated food. Deviations from predicted differences in surveyed literature data are discussed in terms of possible methodological sources of error.

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Résumé Plus de 80% des valeurs de la digestibilité approchée (AD), de l'efficacité de la conversion de la nourriture assimilée en biomasse (ECD) et de l'efficacité de la conversion de l'aliment ingéré (ECI), calculées en termes énergétiques, et obtenus à partir de données sur 65 espèces, sont supérieures aux valeurs des poids secs correspondants (DW): 38 études; valeurs comparatives: AD=139, ECD=128, ECI=169. Les plus importantes différences positive (énergie>valuers DW) sont de 30 (AD, ECD) et de 24 (ECI) centièmes (les différences négatives les plus fortes = 9 (AD), 11 (ECD) et 8 (ECI); ces différences sont moindres pour ECI (71% des différences tombent à 0 et +5 centièmes), et AD (68%), suivi de ECD (seulement 47% tombent entre 0 et +5). Ces différences peuvent varier avec la température, l'alimentation et d'autres facteurs; les différences tendent à croître (particulièrement pour ECD et ECI) quant on les compare plus tard avec des stades plus précoces. Energie > aux valeurs d'efficacité DW sont généralement attendues pour AD par suite du contenu énergétique supérieur de l'aliment à celui des excréments, et pour ECD et ECI par suite du contenu énergétique généralement plus élevé pour la biomasse de l'insecte que pour l'aliment ingéré et assimilé. Les écarts par rapport aux différences prédites dans les données de la littérature examinée sont analysées en considérant les sources possibles d'erreurs méthodologiques.