Development of an enzymatic method for individual separation of banded coral shrimp (Stenopus hispidus) embryos

The outer coat of embryo of Stenopus hispidus may affect cryoprotectant penetration and post‐thaw procedures during cryopreservation studies. The mechanical method for separating embryos of S. hispidus was exhausting and time consuming. The aim of the present study was to compare the effect of different enzymatic isolation techniques to achieve the individual separation of embryos. Embryo clusters were exposed to three enzymes (trypsin, collagenase and hyaluronidase) at different concentrations (0.2–1.6 mg mL^?1) for different treatment times (5–20 min). Morphological appearance and hatching competence of the embryos separated mechanically and enzymatically were assessed. The optimum conditions for the three enzymes were 10 min 0.4 mg mL^?1 trypsin, 10 min 0.8 mg mL^?1 collagenase and 10 min 0.8 mg mL^?1 hyalruonidase and embryo hatching percentages were 43.8 ± 4.2%, 39.9 ± 4.9% and 42.2 ± 3.5% respectively. The enzymatic isolation technique does not affect either morphological appearance or hatching competence of the separated embryos. The method developed in this study will be helpful for future cryopreservation studies on S. hispidus embryos.     

Chitosan-Based Anti-Oxidation Delivery Nano-Platform: Applications in the Encapsulation of DHA-Enriched Fish Oil

Refined cobia liver oil is a nutritional supplement (CBLO) that is rich in polyunsaturated fatty acids (PUFAs), such as DHA and EPA; however, PUFAs are prone to oxidation. In this study, the fabrication of chitosan-TPP-encapsulated CBLO nanoparticles (CS@CBLO NPs) was achieved by a two-step method, including emulsification and the ionic gelation of chitosan with sodium tripolyphosphate (TPP). The obtained nanoparticles were inspected by dynamic light scattering (DLS) and showed a positively charged surface with a z-average diameter of between 174 and 456 nm. Thermogravimetric analysis (TGA) results showed the three-stage weight loss trends contributing to the water evaporation, chitosan decomposition, and CBLO decomposition. The loading capacity (LC) and encapsulation efficiency (EE) of the CBLO loading in CS@CBLO NPs were 17.77–33.43% and 25.93–50.27%, respectively. The successful encapsulation of CBLO in CS@CBLO NPs was also confirmed by the Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD) techniques. The oxidative stability of CBLO and CS@CBLO NPs was monitored by FTIR. As compared to CBLO, CS@CBLO NPs showed less oxidation with a lower generation of hydroperoxides and secondary oxidation products after four weeks of storage. CS@CBLO NPs are composed of two ingredients that are beneficial for health, chitosan and fish oil in a nano powdered fish oil form, with an excellent oxidative stability that will enhance its usage in the functional food and pharmaceutical industries.     

钾肥对富士苹果着色的影响及机理

【目的】钾肥过量是目前苹果果皮发黄、着色不良的重要原因。拟通过田间试验,调查不同钾肥用量在氮、磷、钙、镁不同肥料配合下,对苹果果实色泽、养分元素含量及果皮色素的影响。探讨过量钾肥导致富士苹果果皮发黄的作用机制,为有效预防提供理论依据。【方法】供试树为9年生富士,八棱海棠为砧木,单株产量约60 kg。土壤为中性潮土,中等肥力。根据苹果对营养元素的吸收特性及相互作用原理,参考农民施肥习惯和前人研究结果,试验以N-P、N-P-Mg、N-P-Ca 3种肥料配合为基础,与每株0-2 000 g (0、125、250、500、1 000和2 000 g/株）的K2O 配合施用,测定果实着色度、花青素、叶绿素、类胡萝卜素、果实和土壤中的主要矿质元素含量。【结果】在3种肥料配合中,果实着色度均随钾肥用量的增加呈先提高后降低的变化趋势,N-P-Mg配合果实着色度最好,N-P-Ca配合最差;果实着色最好的钾肥用量,N-P、N-P-Ca配合在250 g/株,N-P-Mg配合延迟至1 000 g/株;N-P配合中钾肥用量与果实K含量、土壤残留K含量呈显著正相关,与果实Mg含量呈极显著负相关;N-P-Mg配合中钾肥用量与果实中矿质元素含量相关性均不显著;N-P-Ca配合中钾肥用量与果实中N含量呈极显著负相关,与果实中Fe含量呈显著正相关,果实中N的均值比N-P处理高37.5%;果皮叶绿素含量随钾肥用量的增加,在N-P配合下呈先提高后降低趋势,在N-P-Mg配合下逐渐降低,而在N-P-Ca配合下呈先降低后提高趋势;果皮类胡萝卜素含量随钾肥用量增加,在N-P-Mg配合下呈先降低后提高的变化趋势,与N-P配合基本一致,而在N-P-Ca配合下基本上呈不断提高的趋势。3种配合的果皮类胡萝卜素含量均在钾肥用量最多时,达到最高,但花青素含量却最低,因此导致果皮发黄,着色不好。【结论】富士苹果着色度随钾肥用量的增加呈先提高后降低趋势;过量施钾促进了果实钾的积累,抑制了镁的吸收,导致果皮中花青素含量降低,类胡萝卜素含量提高,使果皮呈黄色着色不良;配施镁肥使果皮着色最好的K2O用量从250 g/株延迟到1 000 g/株,有效减轻了高量钾肥对苹果着色的不良影响;配施钙肥由于增加了果实中N和Fe的含量,使果皮叶绿素含量提高而花青素含量降低,对果实着色有负作用。本研究结果表明果实色泽是N、P、K、Ca、Mg、Fe、Mn几种养分元素共同作用的结果,增施某种养分时,必需注意保持与其它各元素间的平衡。     

Lipoxygenase from banana leaf: purification and characterization of an enzyme that catalyzes linoleic acid oxygenation at the 9-position

The objective of the present study was to purify and characterize the lipoxygenase (LOX) from banana leaf (Giant Cavendishii, AAA), an unutilized bioresource. LOX was extracted, isolated, and purified 327-fold using 25-50% saturation of ammonium sulfate fractionation, hydroxyapatite column separation, and gel filtration on Superdex 200. The molecular mass of the purified LOX was 85 kDa, K(m) was 0.15 mM, and V(max) was 2.4 microM/min.mg using linoleic acid as substrate. Triton X-100 was required in the extraction medium; otherwise, no LOX activity was detected. LOX activity increased with the concentration of Triton X-100 with an optimum at 0.1%. The optimal pH of the purified LOX from banana leaf was 6.2, and optimal temperature was 40 degrees C. The LOX showed the highest reactivity toward 18:2 followed by 18:3 and 20:4. A very low reaction rate was observed toward 20:5 and 22:6. On the basis of retention time in normal phase HPLC, the products of 18:2 or 18:3 catalyzed by purified LOX were hydroperoxyoctadecadienoic acid or hydroperoxyoctadecatrienoic acid. It seems that 9-LOX is the predominant enzyme in banana leaf. Banada leaf dried at 110 degrees C for 2 h developed algal aroma. Banana leaf extract stored at 10 degrees C for 12 h formed an oolong tea-like flavor. Banana leaf extract reacted with 18:2 or soybean oil pretreated with bacterial lipase produced green and melon-like aroma, whereas the same reaction with 18:3 produced a sweet, fruity, cucumber-like flavor note.     

Production and characterization of human extracellular superoxide dismutase in the methylotrophic yeast Pichia pastoris

Reactive oxygen species are associated with various diseases including cardiovascular diseases, neurological disorders, and pulmonary diseases. Extracellular superoxide dismutase (ECSOD) is an antioxidant enzyme secreted by cells to prevent overproduction of reactive oxygen species. We expressed an ECSOD gene isolated from a human aortic smooth muscle cDNA library in the methylotrophic yeast Pichia pastoris. A synthetic secretion cassette was constructed with the inducible promoter of the alcohol oxidase 1 gene (AOX1) and the yeast alpha-mating factor signal peptide. As much as 25% of the total protein was ECSOD in some transformants grown under inducing conditions. After 36 h of methanol induction, ECSOD was exported into the culture medium at a concentration of approximately 440 mg/L with an antioxidative activity of 760 +/- 20 U/mg ECSOD. Transformed yeast cells were more resistant to heat shock and H(2)O(2) oxidative stress, indicating that the human ECSOD expressed by P. pastoris had multiple biological functions. Our data suggest that the methylotrophic yeast inducible system is suitable for large-scale production of enzymatically active human ECSOD.     

A multi-gene analysis of diversity of bartonella detected in fleas from Algeria

We report the molecular detection of several Bartonella species in 44 (21.5%) of 204 fleas from Algeria collected from 26 rodents and 7 hedgehogs. Bartonella elizabethae and B. clarridgeiae were detected in the fleas collected on hedgehogs. Bartonella tribocorum and B. elizabethae were detected in fleas collected from rats and mice, and sequences similar to an unnamed Bartonella sp. detected in rodents from China were detected in rats as well as a genotype of Bartonella closely related to Bartonella rochalimae detected in fleas collected on brown rats (Rattus norvegicus).     

Effects of intumescent formulation for acrylic-based coating on flame-retardancy of painted red lauan (<Emphasis Type="Italic">Parashorea</Emphasis> spp.) thin plywood

Acrylic emulsion based painted red lauan plywood (Parashorea spp.) is most commonly used for indoor furnishings. This study investigated the enhancement of the fire retardance of painted plywood by interaction among four major components of intumescent formulation: (1) acrylic emulsion resin as binder resin (BR), (2) pentaerythritol as carbonizing substance (CS), (3) melamine as foam producing substance (FPS) and (4) ammonium polyphosphate as dehydrating agent (DA). Effects of changing BR/CS ratios (designated as FRA series) and FPS/DA ratios (designated as FRM series) on flame-retardance of painted plywood were investigated using a cone calorimeter. The intumescent formulation significantly enhanced fire retardancy of painted plywood by exhibiting lower peak release rates and longer times to reach peak release rates, compared with uncoated plywood (UP) panel and plywood panel solely coated with acrylic emulsion resin. Lower BR content in the FRA series and lower FPS content in the FRM series were shown to enhance flame retardancy of painted plywood. The positive correlation between total heat release values under increasing combustion duration and incremental changes of BR and FPS contents in two series further verified the above findings. Consistent with the observed flame retardancy enhancement of painted plywood, lower heats of combustion and weight losses for paints in the FRM series were also identified by oxygen bomb calorimeter measurements and thermogravimetrical analysis. Infrared analysis of the chars indicated the formation of phosphate ester linkages with the lowest BR content in the FRA series and the lowest FPS content in the FRM series showing superior enhancements of flame retardancy for painted red lauan plywood.     

Identification of natural variants affecting chlorophyll content dynamics during rice seedling development

Seedling vigour is an important characteristic in relation to crop growth and yield. Traits such as photosynthetic capacity and chlorophyll content contribute significantly to seedling establishment at the early growth stage in various crop species, including rice. A diverse panel of 227 rice varieties from several countries was evaluated to determine chlorophyll contents at multiple time points during the seedling stage using a soil–plant analysis development (SPAD) meter, a non‐destructive portable device. Using new statistical approaches, several chromosomal regions associated with variations in chlorophyll content in the third leaf at 13, 16 and 19 days after imbibition were detected. A single nucleotide polymorphism (SNP) cluster on the end of chromosome 11 was significantly associated with the onset of leaf senescence. This region was enriched with genes related to cell death and the stress response. We have identified rice germplasm showing delayed‐senescence phenotypes, these could be suitable donors and genetic resources for breeding, and the use of significant SNP markers associated with these traits could enhance the efficiency of their selection in breeding programmes.     

Radial growth of <Emphasis Type="Italic">Cinnamomum kanehirae</Emphasis> Hayata displays a larger temperature sensitivity in dominant than codominant trees

Key message

The radial wood growth curves of Cinnamomum kanehirae Hayata (an endangered species of subtropical Taiwan) exhibit an S shape. The dominant trees displayed a larger radial growth than the codominant trees, and their growth was more sensitive to air temperature.

Context

Knowledge of wood radial growth is important for evaluating the factors that limit tree growth performance. The relevant experiments have mostly been conducted in cold and temperate ecosystems, but rarely in subtropical ecosystems.

Aims

In this study, we aimed to construct a unified radial growth model for Cinnamomum kanehirae Hayata and to identify its sensitivity to temperature.

Methods

The wood radial increments were quantified for 3 years by either pinning or microcoring. The radial wood growth curves were modelled integratively by semiparametric regression and individually by curve fitting. The effects of tree social class, interannual and environmental factors on radial growth were analysed quantitatively.

Results

A unified S-shaped growth model for C. kanehirae was successfully constructed. By including the social class effect, the model was significantly improved. The maximum radial increment (A) was significantly correlated with the maximum growth rate (μ); both A and μ were significantly higher in dominant than in codominant trees. The time-varying radial growth rate was more sensitive to air temperature in dominant than in codominant trees.

Conclusion

Semiparametric models revealed an S-shaped growth curve of C. kanehirae and confirmed the higher temperature sensitivity of dominant trees compared to codominant trees in humid subtropical areas.