Systemic disruption of the homeostasis of transfer RNA isopentenyltransferase causes growth and development abnormalities in Bombyx mori

Isopentenylation at A37 (i⁶A37) of some transfer RNAs (tRNAs) plays a vital role in regulating the efficiency and fidelity of protein synthesis. However, whether insects, which are well known for their highly efficient protein synthesis machinery, employ this regulatory mechanism remains uninvestigated. In the current study, a candidate tRNA isopentenyltransferase (IPT) gene with three alternative splicing isoforms (BmIPT1–BmIPT3) was identified in Bombyx mori (silkworm). Only BmIPT1 could complement a yeast mutant lacking tRNA IPT. Phylogenetic analysis showed that silkworm tRNA IPT is conserved in the Lepidoptera. BmIPT was expressed in all B. mori tissues and organs that were investigated, but was expressed at a significantly higher level in silk glands of the fourth instar compared to the first day of the fifth instar. Interestingly, BmIPT was expressed at a significantly higher level in the domesticated silkworm, B. mori, than in wild Bombyx mandarina in multiple tissues and organs. Knock‐down of BmIPT by RNA interference caused severe abnormalities in silk spinning and metamorphosis. Constitutive overexpression of BmIPT1 using a cytoplasmic actin 4 promoter in B. mori raised its messenger RNA level more than sixfold compared with nontransgenic insects and led to significant decreases in the body weight and cocoon shell ratio. Together, these results confirm the first functional tRNA IPT in insects and show that a suitable expression level of tRNA IPT is vital for silk spinning, normal growth, and metamorphosis. Thus, i⁶A modification at position A37 in tRNA probably plays an important role in B. mori protein synthesis.     

Preparation and characterisation of vancomycin-impregnated gelatin microspheres/silk fibroin scaffold

A type of antibacterial silk fibroin (SF) scaffold was developed and characterised as a potential functional wound dressing for acute trauma treatment. To achieve this, SF solution was mixed with previously fabricated vancomycin (Vm)-loaded gelatin (G) microspheres, followed by a freeze-drying step. Some physical and antimicrobial properties of the prepared Vm/G/SF scaffolds were investigated and the results demonstrated that the average pore size and porosity of the composite scaffold were 60–80?μm and 75%, respectively. The compressive stress and compressive modulus of Vm/G/SF scaffold were 140 and 468?KPa, respectively. Compared with Vm/G microspheres and Vm/SF scaffold, the Vm/G/SF scaffold has slower release rate of Vm. In addition, the Vm release rate of Vm/G/SF scaffold matched well with the degradation rate of SF scaffold. With respect to the antimicrobial effect, the results showed that the Vm/G/SF scaffold had good antimicrobial activity against Staphylococcus aureus (gram-positive), which is a gram-positive bacteria commonly found in infected wounds.     

Evaluation of an antibacterial peptide-loaded amniotic membrane/silk fibroin electrospun nanofiber in wound healing

Antimicrobial peptides (AMPs) are among the compounds that have significant potential to deal with infectious skin wounds. Using wound dressings or skin scaffolds containing AMPs can be an effective way to overcome infections caused by antibiotic-resistant strains. In this study, we developed an amniotic membrane-based skin scaffold using silk fibroin to improve mechanical properties and CM11 peptide as an antimicrobial peptide. The peptide was coated on the scaffold using the soaking method. The fabricated scaffold was characterised by SEM and FTIR, and their mechanical strength, biodegradation, peptide release, and cell cytotoxicity analyses were performed. Then, their antimicrobial activity was measured against antibiotic-resistant strains of Pseudomonas aeruginosa and Staphylococcus aureus. The in vivo biocompatibility of this scaffold was evaluated by subcutaneously implanting it under the skin of the mouse and counting lymphocytes and macrophages in the implanted area. Finally, the regenerative ability of the scaffold was analyzed in the mouse full-thickness wound model by measuring the wound diameter, H&E staining, and examining the expression rate of genes involved in the wound healing process. The developed scaffolds exerted an inhibiting effect on the bacteria growth, indicating their proper antimicrobial property. In vivo biocompatibility results showed no significant count of macrophages and lymphocytes between the test and control groups. The wound closure rate was significantly higher in the wound covered with fibroin electrospun-amniotic membrane loaded with 32 μg/mL CM11, where the relative expression rates of collagen I, collagen III, TGF-β1 and TGF-β3 were higher compared with the other groups.     

重组蛛丝蛋白支架材料在大鼠体内的组织相容性

目的探讨基因重组蛛丝蛋白等4种支架材料在大鼠体内的组织相容性。方法按照生物材料生物学评价试验国家标准进行4种材料的动物体内植入实验,通过大体观察和组织学方法对4种材料进行检测评价。结果4种材料组织相容性优良顺序为pNSR16、pNS2、PVA、pNSR-Z。pNSR16和pNS2支架材料均显示出良好的组织相容性。结论重组蛛丝蛋白有良好的组织相容性,其应用于组织工程的前景是广阔的。     

N-3 fatty acids reduced trans fatty acids retention and increased docosahexaenoic acid levels in the brain

Introduction: The levels of docosahexaenoic acid (DHA, 22:6n-3) and arachidonic acid (AA, 20:4n-6) are critical for the normal structure and function of the brain. Trans fatty acids (TFA) and the source of the dietary fatty acids (FA) interfere with long-chain polyunsaturated fatty acids (LC-PUFA) biosynthesis.

Objectives: The aim of this study was to investigate the effect of TFA supplementation in diets containing different proportions of n-9, n-6, and n-3 FA on the brain FA profile, including the retention of TFA, LC-PUFA levels, and n-6/n-3 PUFA ratios. These parameters were also investigated in the liver, considering that LC-PUFA are mainly bioconverted from their dietary precursors in this tissue and transported by serum to the brain. Also, stearoyl-CoA desaturase-1 (SCD1) and sterol regulatory element-binding protein-1c (SREBP-1c) gene expressions were evaluated.

Methods: Male CF1 mice were fed (16 weeks) diets containing different oils (olive, corn, and rapeseed) with distinct proportions of n-9, n-6, and n-3 FA (55.2/17.2/0.7, 32.0/51.3/0.9, and 61.1/18.4/8.6), respectively, substituted or not with 0.75% of TFA. FA composition of the brain, liver, and serum was assessed by gas chromatography.

Results: TFA were incorporated into, and therefore retained in the brain, liver, and serum. However, the magnitude of retention was dependent on the tissue and type of isomer. In the brain, total TFA retention was lower than 1% in all diets.

Discussion: Dietary n-3 PUFA decreased TFA retention and increased DHA accretion in the brain. The results underscore the importance of the type of dietary FA on the retention of TFA in the brain and also on the changes of the FA profile.     

丝素/壳聚糖/沙蒿籽胶复合水凝胶的制备与表征

目的：制备丝素/壳聚糖/沙蒿籽胶（SF/CS/SA）复合水凝胶,并进行性能表征和进一步优化,为制备性能优异的神经移植物提供依据。方法：采用溶胶-凝胶方法制备SF/CS/SA复合水凝胶,根据成胶效果进行初步筛选,然后对筛选出的样品进行成胶时间、孔隙率、溶胀率、降解残留率和杨氏模量分析。结果：（1）通过丝素、壳聚糖和沙蒿籽胶的不同比例可以制备出SF/CS/SA复合水凝胶,其中9组成胶完整且稳定,编号A-I,进行后续实验。（2）A、B、C、G、H、I组成胶时间较快,且成胶效果好;复合水凝胶的孔隙率主要集中60%和80%附近,溶胀率变化与孔隙率呈正相关;A、B、G、H、I组降解性能良好;力学性能测定发现A、B组柔性较好,而C、G、H组脆性较好。结论：筛选出成胶时间较快、溶胀性能较好、降解速度较慢、力学性能优良的A、B、G、H组丝素/壳聚糖/沙蒿籽胶复合水凝胶,为制备性能优异的神经移植物提供重要参考。     

取向性生物导电型材料的制备及对神经再生的影响*

目的：通过不同方法研制出取向性静电纺丝复合材料,确定材料与神经再生之间的构效关系,为理想的神经再生植入装置的设计提供扎实的理论基础,为周围神经缺损的修复提供更有效的方法。方法：本实验通过切片法结合静电纺丝及3D打印等技术制备具有取向性的丝素（silk fibroin,SF）/石墨烯（Graphene,Gr）复合材料。显微镜观察材料表面结构形貌。电化学分析提供复合材料的电活性表征。细胞共培养观察该复合材料对神经细胞的影响。结果：不同方法制备的不同取向性复合材料具有不同的导电性,其中3D打印生成取向性的静电纺丝复合材料具有良好的导电性,并且通过体外细胞实验表明其支持培养的施万细胞的存活和生长,揭示其具有良好的生物相容性。结论：结果均表明3D打印取向性复合材料结合了优秀的导电性能与良好的生物相容性等性质,很好地模拟了神经发育的天然神经细胞微环境。     

Thermal conditions during juvenile development affect adult dispersal in a spider

Understanding the causes and consequences of dispersal is a prerequisite for the effective management of natural populations. Rather than treating dispersal as a fixed trait, it should be considered a plastic process that responds to both genetic and environmental conditions. Here, we consider how the ambient temperature experienced by juvenile Erigone atra, a spider inhabiting crop habitat, influences adult dispersal. This species exhibits 2 distinct forms of dispersal, ballooning (long distance) and rappelling (short distance). Using a half-sib design we raised individuals under 4 different temperature regimes and quantified the spiders' propensity to balloon and to rappel. Additionally, as an indicator of investment in settlement, we determined the size of the webs build by the spiders following dispersal. The optimal temperature regimes for reproduction and overall dispersal investment were 20 °C and 25 °C. Propensity to perform short-distance movements was lowest at 15 °C, whereas for long-distance dispersal it was lowest at 30 °C. Plasticity in dispersal was in the direction predicted on the basis of the risks associated with seasonal changes in habitat availability; long-distance ballooning occurred more frequently under cooler, spring-like conditions and short-distance rappelling under warmer, summer-like conditions. Based on these findings, we conclude that thermal conditions during development provide juvenile spiders with information about the environmental conditions they are likely to encounter as adults and that this information influences the spider's dispersal strategy. Climate change may result in suboptimal adult dispersal behavior, with potentially deleterious population level consequences.     

Mechanical property and biological performance of electrospun silk fibroin-polycaprolactone scaffolds with aligned fibers

The mechanical strength, biocompatibility, and sterilizability of silk fibroin allow it to be a possible candidate as a natural bone regenerate material. To improve mechanical character and reinforce the cell movement induction, silk fibroin (SF)–polycaprolactone (PCL) alloy was fabricated by electrospinning techniques with a rotating collector to form aligned fibrous scaffolds and random-oriented scaffolds. The scanning electron microscope image of the scaffold and the mechanical properties of the scaffold were investigated by tensile mechanical tests, which were compared to random-oriented scaffolds. Furthermore, mesenchymal stem cells were planted on these scaffolds to investigate the biocompatibility, elongation, and cell movement in situ. Scanning electron microscopy shows that 91% fibers on the aligned fibroin scaffold were distributed between the dominant direction ±10°. With an ideal support for stem cell proliferation in vitro, the aligned fibrous scaffold induces cell elongation at a length of 236.46 ± 82 μm and distribution along the dominant fiber direction with a cell alignment angle at 6.57° ± 4.45°. Compared with random-oriented scaffolds made by artificial materials, aligned SF-PCL scaffolds could provide a moderate mesenchymal stem cell engraftment interface and speed up early stage cell movement toward the bone defect.