Preparation and Characterization of a Silk Fibroin/Polyurethane Fiber Blend Membrane Containing Actinomycin X2 with Excellent Mechanical Properties and Enhanced Antibacterial Activities

Development of suitable antimicrobial biomaterials for hygienic wound dressing and healing is an important requirement for medical application. Durable mechanical properties increase the application range of biomaterial in different environmental and biological conditions. Due to the inherent brittleness of silk fibroin (SF), polyurethane fiber (PUF) was used to modify SF containing actinomycin X2 (Ac.X2) to prepare silk fibroin@actinomycin X₂/polyurethane fiber (ASF/PUF) blend membranes. The ASF/PUF blend membrane was developed by solution casting method. Incorporation of PUF improved the flexibility of material and introduction of Ac.X2 has increased antibacterial activity of materials. Excellent mechanical properties (tensile strength up to 25.7 MPa and elongation at break up to 946.5 %) of 50 % SF+50 % PUF blend membrane were proved by tensile testing machine. FT-IR spectra, TGA, contact angle and DMA were tested to prove the blend membrane's physico-chemical characteristics. ASF/PUF blend membrane displayed satisfactory antibacterial activity against S. aureus, and the cytotoxicity tests showed that the blend membrane has better biosafety compared to directly applied Ac.X2 in soluble form. These results suggest that the modification of SF through PUF for development of flexible antibacterial membranes has great potential application value in the field of silk-like material fabrication.     

In Silico Design of an Oseltamivir Derivative with Increased Affinity against Wild-Type and Mutant Variants of Neuraminidase and Hemagglutinin of Influenza A H1N1 Virus

Antiviral resistance has turned into a world concern nowadays. Influenza A H1N1 emerged as a problem at the world level due to the neuraminidase (NA) mutations. The NA mutants conferred resistance to oseltamivir and zanamivir. Several efforts were conducted to develop better anti-influenza A H1N1 drugs. Our research group combined in silico methods to create a compound derived from oseltamivir to be tested in vitro against influenza A H1N1. Here we show the results of a new compound derived from oseltamivir but with specific chemical modifications, with significant affinity either on NA (in silico and in vitro assays) or HA (in silico) from influenza A H1N1 strain. We include docking and molecular dynamics (MD) simulations of the oseltamivir derivative at the binding site onto NA and HA of influenza A H1N1. Additionally, the biological experimental results show that oseltamivir derivative decreases the lytic-plaque formation on viral susceptibility assays, and it does not show cytotoxicity. Finally, oseltamivir derivative assayed on viral NA showed a concentration-dependent inhibition behavior at nM, depicting a high affinity of the compound for the enzyme, corroborated with the MD simulations results, placing our designed oseltamivir derivative as a potential antiviral against influenza A H1N1.     

Antibacterial Compound Isolation and Characterization from the Plant Cynotis axillaris Schult

A novel flavone glycoside was isolated from the methanolic extract of Cynotis axillaris Schult. Various analysis and characterization techniques were used to determine its structure and properties. The compound exhibited a melting point range of 231–232 °C and had a molecular formula of C₂₇H₃₀O₁₄. Several spectral characterization techniques were employed to establish the isolated compound's structure. These included UV-visible spectroscopy, FT-IR, LC-ESI-MS, and NMR spectroscopy. Based on these analyses, the structure of the isolated compound was determined to be 5,7,4’-trihydroxyflavone-8-α-L-rhamnopyranoside-4’-O-β-D-galactopyranosyl. This structure indicates that it is a flavone glycoside consisting of a flavone (5,7,4’-trihydroxyflavone) moiety attached to a sugar molecule (galactopyranosyl) at position 4’, which further bears a rhamnose group at position 8 of the flavone. In addition, to the structural characterization, the compound also demonstrated significant antibacterial efficacy against various bacterial pathogens, including Gram-positive bacteria such as Bacillus subtilis MTCC441 and Gram-negative bacteria such as Escherichia coli MTCC1098, Proteus vulgarize MTCC426, and Salmonella Typhimurium MTCC3224. The antimicrobial activity was evaluated by measuring the zone of inhibition in millimetres, which provides an indication of the compound's ability to inhibit bacterial growth. The study successfully identified and characterized a novel flavone glycoside from Cynotis axillaris Schult. and its antimicrobial activity.     

Comparison of mechanistic and hybrid modeling approaches for characterization of a CHO cultivation process: Requirements,pitfalls and solution paths

Despite the advantages of mathematical bioprocess modeling, successful model implementation already starts with experimental planning and accordingly can fail at this early stage. For this study, two different modeling approaches (mechanistic and hybrid) based on a four-dimensional antibody-producing CHO fed-batch process are compared. Overall, 33 experiments are performed in the fractional factorial four-dimensional design space and separated into four different complex data partitions subsequently used for model comparison and evaluation. The mechanistic model demonstrates the advantage of prior knowledge (i.e., known equations) to get informative value relatively independently of the utilized data partition. The hybrid approach displayes a higher data dependency but simultaneously yielded a higher accuracy on all data partitions. Furthermore, our results demonstrate that independent of the chosen modeling framework, a smart selection of only four initial experiments can already yield a very good representation of a full design space independent of the chosen modeling structure. Academic and industry researchers are recommended to pay more attention to experimental planning to maximize the process understanding obtained from mathematical modeling.     

Purification of antibacterial attacins induced in Hyalophora cecropia pupae immunized with Enterobacter cloacae C7-501 using ion-exchange chromatography,hydrophobic interaction chromatography,and Rotofor® isoelectric focusing

Hyalophora cecropia pupae were infected by Enterobacter cloacae C7-501 to induce antibacterial attacins for purification. The induction of attacins in immunized pupae was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Ion-exchange chromatography (IEC), hydrophobic interaction chromatography (HIC), and Rotofor® isoelectric focusing (ISEF) were applied to isolate attacins from the hemolymph. IEC separated attacins from most hemolymph proteins, but the fractions containing attacins also had other proteins of 20 and 64 kDa in length. In IEC, attacin was eluted with ~0.2 M NaCl. The best conditions for IEC were pH 9, flow rate of 2 mL/min, with step elution (0.025, 0.05, 0.075, 0.1, 0.2, 0.4 and 1.0 M NaCl). In HIC, most other proteins were eluted with the ammonium persulfate treatment. HIC isolated attacin proteins under hydrophobic conditions, at ~50% EtOH. However, the fraction with attacins also contained other proteins. The Rotofor® ISEF produced fractions containing attacins at isoelectric points ranging between 5.7 and 8.3. However, non-specific proteins were detected in the fraction samples, and the recovery of attacins was low. The purification efficiency of ISEF was lower than IEC and HIC. In this study, the expression of attacins was induced in H. cecropia pupae infected with E. cloacae C7-501, and attacins could be purified by IEC and ISEF. Overall, IEC provided better separation of attacins from the hemolymph of H. cecropia pupae immunized with E. cloacae bacteria than HIC and Rotofor® ISEF.     

Search for substrate-based inhibitors fitting the S2' space of malarial aspartic protease plasmepsin II.

Plasmepsin (Plm) has been identified as an important target for the development of new antimalarial drugs, since its inhibition leads to the starvation of Plasmodium falciparum. A series of substrate-based dipeptide-type Plm II inhibitors containing the hydroxymethylcarbonyl isostere as a transition-state mimic were synthesized. The general design principle was provision of a conformationally restrained hydroxyl group (corresponding to the set residue at the P2' position in native substrates) and a bulky unit to fit the S2' pocket.     

Population genetic structure of the butterfly Melitaea didyma (Nymphalidae) along a northern distribution range border

The population genetic structure of the butterfly Melitaea didyma was studied along the northern distribution range border in Central Germany by means of allozyme electrophoresis. Individuals were sampled from a total of 21 habitat patches from four regions, and two provinces. Sampling was designed to estimate local vs. regional differentiation. High levels of variability were found, H _e= 0.14–0.21. The mean expected sample heterozygosity from one region, Mosel, was significantly lower than from the Hammelburg region, H _e= 0.17 and 0.19, respectively. Two hierarchical levels of genetic differentiation were found. Within regions individuals sampled from different patches behaved as belonging to one population with high levels of gene flow (Hammelburg F _ST= 0.015, Mosel F _ST= 0.044), though local isolation barriers did create a substructuring of these populations. The inbreeding coefficients, F _IS, were constant over all sample levels, suggesting a similar distribution of habitat patches within regions. Between regions gene flow was limited. An isolation by distance analysis indicated that the hierarchical structure, at the provincial level, may be breaking down due to isolation of regional populations. A more general observation was that the sampling design may greatly have influenced the estimation of genetic differentiation. Depending on which samples were included, overall F _ST estimates ranged from 0.059–0.090.     

饱和试验设计资料的逐步回归分析

对饱和试验设计资料,一次最小二乘分析是不合理的,采用逐步回归分析法,在选择适当的理论F值前提下,不仅可以检验每个回归系数的显著性,而且能够获得最优回归方程．     

适用于获取最优化配方的一种算法

本文应用印楝种仁提取物（F3）与敌敌畏混配为例,以斜纹夜蛾（Spodopteralitura）为目标害虫,介绍一种适用于获取最优化配方的算法,在二次通用回归旋转组合设计的基础上,经参数辨识,获取二次回归方程,经失拟性、回归显著性检验,本方程基本能够反映杀虫剂用量与斜纹夜蛾幼虫死亡机率值之间的关系．在害虫防治实践中,要求在防治费用最小的基础上,目标害虫有最大的死亡率.因此,以防治目标害虫的费用作为优化算法的目标函数,以害虫死亡机率值最大作为约束条件,有如下的一组优化算式为目标函数约束条件式中a1,a2分别为参试杀虫剂1,2最低用量,b1,b2则为相应的最高用量．C1,C2分别为杀虫剂1,2的单价,N1,N2为杀虫剂l,2的用量．Y为目标害虫死亡机率值回归方程．本文所依据的试验设计中,以对数函数关系变换编码值与使用浓度之间的关系,所以应用拉格朗日求极值原理求取最优化配方．由计算所得的混配比例与其他方法所获结果一致．     

多元方差分析中的单一自由度独立比较

本文根据一元方差分析的单一自由度比较原理提出了多元方差分析中的单一自由度比较方法,以用于各种平衡试验设计中具多个变量的处理间多重比较．