Cellulose-based Polymeric Liquid Crystals as a Biomimetic Modifier for Suppressing Protein Adsorption

A novel biomimetic protein-resistant modifier based on cellulose-based polymeric liquid crystals was described（PLCs）. Two types of PLCs of propyl hydroxypropyl cellulose ester（PPC） and octyl hydroxypropyl cellulose ester（OPC） were prepared by esterification from hydroxypropyl cellulose, and then were mixed with polyvinyl chloride and polyurethane to obtain composite films by solution casting, respectively. The surface morphology of PLCs and their composite films were characterized by polarized optical microscopy（POM） and scanning electron microscopy（SEM）, suggesting the existence of microdomain separation with fingerprint texture in PLC composite films. Water contact angle measurement results indicated that hydrophilicity of PLC/polymer composite films was dependent on the type and content of PLC as well as the type of matrix due to their interaction. Using bovine serum albumin（BSA） as a model protein, protein adsorption results revealed that PLCs with protein-resistant property can obviously suppress protein adsorption on their composite films, probably due to their flexible LC state. Moreover, all PLCs and their composites exhibited non-toxicity by MTT assay, suggesting their safety for biomedical applications.

Protein Drug-Loaded Polymeric Nanoparticles

Considerable interest and research have focused on the administration of therapeutic proteins. For delivery of therapeutic proteins, bioavailability and stabilization of protein drugs to maintain therapeutically acceptable levels is an important challenge in clinical trials. To overcome these challenges, polymeric nanoparticles have become one of the best methods for protein delivery. In this review, we summarize the current available polymeric nanoparticles designed for protein delivery, current status, and advantages of protein delivery systems.

Transfer and Reactivity of Hydrogen Sulfide with Immobilized Hemeproteins in Polymeric Matrix

Hydrogen sulfide (H2S) has been related to be toxic and to have a role in human physiological functions. Therefore, there is a necessity to comprehend ways to scavenger hydrogen sulfide from different media. Here, we used recombinant metaquo-Hemoglobin I (metHbI) from Lucina pectinata and metaquo-myoglobin (metMb) encapsulated in the tetramethyl orthosilicate gel (TMOS), to facilitate the understanding of H2S transfer toward these metaquo-hemeproteins. In this sol-gel environment, metHbI binds and releases H2S with rate constants of 0.0597 M-1·s-1 and 6.67 × 10-5 s-1, respectively. The process generates an H2S affinity constant (kon/koff) of 8.9 × 102 M-1, which is 107 lowers than the analogous constant in solution (6.3 × 109 M-1). Although the H2S koff for the rHbI-H2S complex is almost similar with both sol-gel and solution. To further understand how the H2S koff from rHbI-H2S in solution (5 μM) is influenced by the protein concentration gradient, metHbI and metMb (25 μM) encapsulated in TMOS sol-gel. Under these circumstances, the H2S transfer from a solution of the rHbI-H2S complex to encapsulated hemeprotein resulted in koff values of 1.90 × 10-4 s-1, and 2.09 × 10-4 s-1 leading to the formation of rHbI-H2S and Mb-H2S species, respectively. The results suggest that the: 1) extreme ionic TMOS construct limits the H2S pathways to reach the hemeprotein active center, 2) possible interaction with metHbI hydrophilic forces increases the hydrogen bonding networking and decreases the H2S association constant, 3) hemeproteins concentration gradients between solution and sol-gels also influence its hydrogen sulfide transfer. In the presence of oxygen or hydrogen peroxide metMb generated a mixture of Mb-H2S and sulfmyoglobin derivative, while encapsulated metHbI reaction did not produce the sulfheme species. Consequently, the results show that metHbI encapsulated in TMOS is an excellent trap for H2S from solution or gas media.

Electrochemical Impedance Analysis of Biofunctionalized Conducting Polymer-Modified Graphene-CNTs Nanocomposite for Protein Detection

We report an electrodeposited poly(pyrrole-co-pyrrolepropylic acid) copolymer modified electroactive graphene-carbon nanotubes composite deposited on a glassy carbon electrode to detect the protein antigen(cTnI). The copolymer provides pendant carboxyl groups for the site-specific covalent immobilization of protein antibody, antitroponin I. The hybrid nanocomposite was used as a transducer for biointerfacial impedance sensing for cTnI detection.The results show that the hybrid exhibits a pseudo capacitive behaviour with a maximum phase angle of 49° near 1 Hz,which is due to the inhomogeneous and porous structure of the hybrid composition. The constant phase element of copolymer is 0.61(n = 0.61), whereas, it is 0.88(n = 0.88) for the hybrid composites, indicating a comparatively homogeneous microstructure after biomolecular functionalization. The transducer shows a linear change in charge transfer characteristic(R_(et)) on cTnI immunoreaction for spiked human serum in the concentration range of 1.0 pg mL^(-1)–10.0 ng mL^(-1). The sensitivity of the transducer is 167.8 ± 14.2 Ω cm^2 per decade, and it also exhibits high specificity and good reproducibility.

Molecularly Imprinted Polymer Based on GO-QDs as Enhanced Fluorescent Nanoscale Device for Specific Recognition of Target Protein

In this work the enhanced molecularly imprinted optosensing material based on graphene oxide-quantum dots （ GO- QDs） was synthesized for highly selective and sensitive specific recognition of the target protein, bovine serum albumin （BSA）. Here, GO was introduced to enhance the efficiency of mass-transfer in recognition of target protein. Molecularly imprinted polymer coated GO-QDs using BSA as template （BMIP-coated GO-QDs ） exhibited a fast mass-transfer speed, which could be ascribed to the high volume of efficient surface area and high target recognition efficiency of the synthesized nanoscale device. Under optimal conditions, it was found that the BSA as target protein could remarkably quench the relative fluorescence intensity of BMIP- coated GO-QDs linearly in a concentration-dependent manner that was best described by a Stern-Volmer equation. The Ksv （Stern- Volmer constant） for template BSA was much higher than bovine hemoglobin （BHb） and lysozyme （Lyz）, implying a highly selective recognition ability of the BMIP-coated GO-QDs to BSA. This enhanced fluorescent nanoscale device may provide opportunities to develop a system that is efficient and effective and has potential in the design of highly effective fluorescent receptor for recognition of target protein in Droteomics studies.

A strong adhesive block polymer coating for antifouling of large molecular weight protein

Some proteins secreted by microorganisms have large molecular weights. We report here an approach to prepare coating by multilayer polymers for antifouling of proteins, especially the proteins with a large molecular weight.Stainless steel was used as the model substrate. The substrate was first coated with a hybrid polymer film, which was formed by simultaneous hydrolytic polycondensation of 3-aminopropyltriethoxysilane and polymerization of dopamine(HPAPD). After grafting the macroinitiator 2-bromoisobutyryl bromide, the block polymer brushes PMMA-b-PHEMA were grafted. Three proteins were used to test protein adsorption and antifouling behavior of the coating, including recombinant green fluorescent(54 k Da), recombinant R-transaminase(2 × 90 k Da), and recombinant catalase(4 × 98 k Da). It is demonstrated that the block polymer brushes not only can prevent the adsorption of small molecular weight proteins, but also can significantly reduce the adsorption of the large molecular weight proteins.

不同外源Cd(Ⅱ)对Pseudomonas aeruginosa EPS的胁迫效应--产量、组分、吸附特性变化及其机制

以CdCl_(2)、CdSO_(4)和Cd(NO_(3))_(2)为胁迫因子,比较了不同阴离子在胁迫/诱导过程中对Pseudomonas aeruginosa(P.aeruginosa)胞外聚合物(EPS)的产量和特性的影响以及对Cd(II)吸附性能的变化.研究发现,当Cd(NO_(3))_(2)胁迫/诱导浓度为50mg/L时,EPS产量达到214.91mg/g VSS,其中蛋白质含量达到136.75mg/g VSS,较胁迫/诱导前增加了409.31%.此时EPS的吸附性能最好,对Cd(II)的平衡吸附量最大,较胁迫/诱导前增加了近一半.HPLC分析表明,EPS蛋白质中氨基酸的变化与EPS的吸附特性密切相关.3D-EEM、XPS和FTIR结果表明,EPS中C=O、C-N和N-H含量增加,有利于重金属的结合.竞争吸附实验发现,EPS对Cd(II)的亲和力高于Zn(II).

牡蛎疱疹病毒结构蛋白真核表达系统构建及多聚化特性

牡蛎疱疹病毒(OsHV-1)在全球范围内导致牡蛎、扇贝与蚶类的大规模死亡,成为双壳贝类养殖产业的重要威胁。为了解OsHV-1的结构与致病机制。本研究利用人胚胎肾细胞(HEK293t),构建OsHV-1主要核衣壳蛋白(ORF104和ORF33)的真核表达系统,并对ORF104和ORF33潜在相互作用进行分析。实验首先通过特异性PCR扩增技术得到orf 104和orf 33的基因序列,根据其编码蛋白的理化性质、跨膜区与三维结构等生物信息学分析结果,选择pCDNA3.1(+)构建两种基因的重组表达质粒。重组质粒经大肠杆菌扩增、提取后,利用转染试剂Lipo8000™将pCDNA3.1(+)-orf 104与pCDNA3.1(+)-orf 33分别单独或共转染至HEK293t。然后,将转染后的细胞培养18 h后裂解收集蛋白。最后利用蛋白免疫印迹(Western blot,WB)与负染电镜检测两种目的蛋白的表达情况。结果显示,实验成功构建了OsHV-1衣壳蛋白ORF104和ORF33的重组表达质粒载体,通过真核细胞表达得到大小约为135与35 ku的目的蛋白。研究表明,表达质粒可在真核表达系统中实现蛋白单独转染与共转染,共转染蛋白间可能存在相互作用的趋势并形成多聚体,其中,ORF33自身即可形成分子质量不同的多聚体。本研究首次利用真核表达系统开展OsHV-1关键结构蛋白的表达,为进一步开展该病毒结构蛋白功能与互作,以及病毒入侵机制研究奠定基础。

磁性核壳微球的制备及其性能

以苯乙烯、甲基丙烯酸为单体,在偶氮二异丁腈引发下通过分散聚合合成了羧基化纳米单分散共聚微球。用化学沉淀法在共聚微球表面生成均匀分散的纳米四氧化三铁壳层,采用FTIR、SEM、DLS、HR-TEM、XRD、XPS及VSM对微球进行形貌及结构表征。结果显示微球均有良好单分散性且在水相中分散性良好。以BSA为模型研究了核壳微球蛋白分离性能,在体系pH值为4.7时微球对蛋白分离效果最佳,吸附等温线符合Langmuir等温吸附模型,微球的蛋白分离性能达到239.5 mg/g。

Glycine betaine modulates extracellular polymeric substances to enhance microbial salinity tolerance

High salinity inhibits microbial activity in the bioremediation of saline wastewater.To alleviate osmotic stress,glycine betaine(GB),an osmoprotectant,is added to enhance the secretion of extracellular polymeric substances(EPS).These EPS are pivotal in withstanding environmental stressors,yet the intricate interplay between GB supplementation and microbial responses through EPS modificationsdencompassing composition,molecular architecture,and electrochemical featuresdremains elusive in hypersaline conditions.Here we show microbial strategies for salinity endurance by investigating the impact of GB on the dynamic alterations of EPS properties.Our findings reveal that GB supplementation at 3.5%salinity elevates the total EPS(T-EPS)content from 12.50±0.05 to 24.58±0.96 mg per g dry cell weight.The observed shift in zeta potential from-28.95 to-6.25 mV at 0%and 3.5%salinity,respectively,with GB treatment,indicates a reduction in electrostatic repulsion and compaction.Notably,the EPS protein secondary structure transition from b-sheet to a-helix,with GB addition,signifies a more compact protein configuration,less susceptible to salinity fluctuations.Electrochemical analyses,including cyclic voltammetry(CV)and differential pulse voltammetry(DPV),reveal GB's role in promoting the release of exogenous electron shuttles,such as flavins and c-type cytochromes(c-Cyts).The enhancement in DPV peak areas(Q_(DPV))with GB addition implies an increase in available extracellular electron transfer sites.This investigation advances our comprehension of microbial adaptation mechanisms to salinity through EPS modifications facilitated by GB in saline habitats.

甲鱼蛋白源寡肽体外对微管蛋白聚合-解聚调控模式的影响

为探讨甲鱼蛋白源寡肽对肿瘤细胞微管蛋白活性的调控机制,本研究以前期获得的甲鱼蛋白源三条寡肽DEADLLA(D-7-A)、EAGVNDW(E-7-W)、GSISSGQV(G-8-V)为研究对象,评价其对微管蛋白聚合-解聚的影响,解析其与微管蛋白的作用模式,阐释其诱导肿瘤细胞凋亡作用的机制。结果表明,3条寡肽与长春新碱相似,均为微管蛋白聚合的抑制剂,IC_(50)值分别为3.72、5.01、5.95μmol/L。分子对接结果显示,D-7-A与α-微管蛋白4X1I活性中心结合位点为Ser178和Thr179;E-7-W与活性中心结合位点为Gln15、Thr225、Tyr224、Tyr210、Arg214;G-8-V与活性中心结合位点为Gln11、Ser178、Asp329,与α-微管蛋白具有较强的作用力。细胞实验结果表明,D-7-A、E-7-W、G-8-V均对A549细胞具有抑制作用,且具有剂量依赖性,其IC_(50)值分别为2003±72、1877±102和1789±137μmol/L。G-8-V对A549细胞的抑制效果最强,且具有诱导A549细胞凋亡的作用,并随着药物处理时间的增加,凋亡率明显提高,其作用机制是通过影响α-微管蛋白的价键,抑制微管蛋白的聚合,诱导细胞凋亡。

G蛋白偶联受体多聚化的研究进展

G蛋白偶联受体(G Protein-Coupled Receptor,GPCR)是真核细胞膜表面受体家族中规模最大且种类最多的一类受体蛋白,常以单体、同型或异型二聚体等形式发挥作用,具有良好的成药性。最近的研究发现,GPCR多聚化有助于GPCR的表达、成熟,并影响其与配体互作以及下游信号转导等过程。本文综述了GPCR多聚体在蛋白分选、配体调节、信号转导和内化过程中的作用,以及在某些疾病发生发展中的病理与生理作用机制,并探讨了GPCR多聚体研究的技术方法、目前存在的问题和未来发展方向,这对研发靶向GPCR多聚体的药物具有重要意义。

CMC-MMA-AA纳米乳液的制备及其抗蛋白质吸附性能的研究

本文以羧甲基纤维素(CMC)为乳化剂,甲基丙烯酸甲酯(MMA)和丙烯酸(AA)为单体,H_(2)O_(2)+Fe^(2+)为引发剂,采用乳液聚合技术制备了CMC-MMA-AA纳米乳液。通过SEM、DLS、TG、IR对乳胶粒的理化性质进行表征,并采用BLI、SDS-PAGE等方法验证纳米粒子的抗蛋白质吸附性能。结果表明:使用羧甲基纤维素包被后的聚甲基丙烯酸甲酯-丙烯酸纳米粒子粒径大小均一,粒径分布较窄,且具备良好的抗蛋白质吸附性能。

Effect of proteins,polysaccharides,and particle sizes on sludge dewaterability

Four batch experiments of hydrolysis and acidification were carried out to investigate the distributions of proteins （PN） and polysaccharides （PS） in the sludge, the PN/PS ratio, the particle sizes, and their relationship with sludge dewaterability （as determined by capillary suction time, CST）. The sludge flocs were stratified through centrifugation- and ultrasound-based method into four fractions： （1） slime, （2） loosely bound extracellular polymeric substances （LB-EPS）, （3） tightly bound EPS （TB-EPS）, and （4） pellet. The results showed that PN was mainly partitioned in the pellet （80.7%） and TB-EPS （9.6%） fractions, while PS distributed evenly in the four fractions. During hydrolysis and acidification, PN was transferred from the pellet and TB-EPS fractions to the slime fraction, but PS had no significant transfer trends. The mean particle sizes of the sludge flocs decreased with hydrolysis and acidification. The pH had a more significant influence on the dewaterability of sludge flocs than temperature. Sludge dewaterability during hydrolysis and acidification processes greatly deteriorated from 9.7 s at raw sludge to 340-450 s under alkaline conditions. However, it was just slightly increased under acidic conditions. Further investigation suggested that CST was affected by soluble PN, soluble PN/PS, and particle sizes of sludge flocs, but was affected slightly by total PN, PS, or PN/PS in the whole sludge flocs and other fractions （except slime）.

Design and Fabrication of Superparamaganitic Hybrid Microspheres for Protein Immobilization

Superparamagnetic poly（styrene）-co-poly（2-acrylanmido-2-methyl propanesulfonic acid） （PSt-co-PAMPS） and poly（methylmethacrylate）-co-poly（glycidyl methacrylate） （PMMA-co-PGMA） microspheres with mean size of 170 nm were prepared by emulsion polymerization in the presence of oleic acid-coated Fe3O4 nanoparticles. The structures, morphologies, diameter and diameter distribution of the as-prepared microspheres were identified by Fourier transform infrared spectroscopy （FT-IR） and transmission electron microscopy （TEM）. The saturation magnetizations of PSt-co-PAMPS and PMMA-co-PGMA microspheres are 21.94 and 25.07 emu/g, respectively. The as-synthesized magnetic microspheres were used for immobilization of Bovine serum albumin （BSA） by physical interaction and covalent interaction respectively. The equilibrium amount of BSA immobilized onto PMMA-co-PGMA microspheres was 86.48 mg/g microspheres in 90 min, while on PSt-co-PAMPS microspheres was 59.62 mg/g microspheres in 120 min.

Plasma graft of poly(ethylene glycol) methyl ether methacrylate (PEGMA) on RGP lens surface for reducing protein adsorption

Poly（ethylene glycol） methyl ether methacrylate（PEGMA） was grafted on fluorosilicone acrylate rigid gas permissible contact lens surface by means of argon plasma induced polymerization to improve surface hydrophilicity and reduce protein adsorption.The surface properties were characterized by contact angle measurement,x-ray photoelectron spectroscopy（XPS） and atomic force microscopy respectively.The surface protein adsorption was evaluated by lysozyme solution immersion and XPS analysis.The results indicated that a thin layer of PEGMA was successfully grafted.The surface hydrophilicity was bettered and surface free energy increased.The lysozyme adsorption on the lens surface was reduced greatly.

Interpenetrated Chitosan-Poly(Acrylic Acid-Co-Acrylamide) Hydrogels. Synthesis, Characterization and Sustained Protein Release Studies

Interpenetrated polymer networks of chitosan (CHI), polyacrylic acid (PAA) and polyacrylamide (PAM) were prepared by free radical polymerization. These hydrogels were either washed with double distilled water (CHI/PAA/PAM) A or hydrolyzed with 1M sodium hydroxide (NaOH), (CHI/PAA/PAM) S. Both types of hydrogels were characterized by infrared spectroscopy, microstructural techniques and compressive mechanical testing. Finally, hydrogels were loaded with bovine serum albumin (BSA) and release followed at different pHs. Infrared spectra analysis showed correspondence between hydrogels and monomer feed compositions. Hydrolyzed hydrogels, had increased water content and pH swelling dependence. Compression modulus of swelled hydrolyzed hydrogels decreased with increasing equilibrium water content. Higher BSA loadings were achieved on hydrolyzed hydrogels due to their high water content and porosity. Protein release from hydrogels was low (≤ 20% after 10 hours) at pH 1.2, but sustained release was observed at pH 6.8 and 7.4. The integrity of the protein released at 6.8 and 7.4 by hydrolyzed hydrogels was unaffected. The hydrogles showed no cytotoxic effects on human skin dermal fibroblasts as determined by MTT assay except for two compositions of (CHI/PAA/PAM) A samples, which after seven days presented a viability lower than 80% respect to the control.

Effects of Propofol combined with remifentanil on the levels of MBP,NSE and S100B protein,D-D and inflammatory factors in patients with acute craniocerebral trauma

Objective: To investigate the effects of Propofol combined with remifentanil on serum levels of MBP, NSE and S100B protein, D-D and inflammatory factors in patients with acute craniocerebral trauma. Methods: A total of 100 patients were selected with traumatic brain injury who underwent emergency surgery from August 2014 to May 2017 in our hospital, then randomly divided them into the control group and the experimental group, 50 cases each. The control group received isoflurane combined with remifentanil to maintain anesthesia, and the experimental group received propofol and remifentanil to maintain anesthesia. The inflammatory factors and the levels of MBP, NSE, S100B and D-D in the two groups before and after anesthesia (T0), 1H (T1) and postoperative 1H (T2) were detected and compared. Results: There was no significant difference between the two groups in the levels of TNF-α. The serum level of hs-CRP in two groups of T1, T2 increased significantly, the difference was statistically significant compared with T0, in the experimental group, serum level of hs-CRP at T1 and T2 was significantly higher than the control group, the difference was statistically significant. Conclusion: Propofol combined with remifentanil anesthesia for acute craniocerebral trauma can maintain the balance of inflammatory cytokine levels during the perioperative period, inhibit the elevation of serum MBP, NSE, S100B protein and D-D levels, reduce brain cell damage. It has a good protective effect on brain cells and is worthy of clinical application.

Polymer Nanoparticles Prepared by Supercritical Carbon Dioxide for in Vivo Anti-cancer Drug Delivery

A new approach for producing polymer nanoparticles made of bovine serum albumin-poly(methy methacrylate) conjugate by precipitating in supercritical CO2 is reported. The nanoparticles were loaded with the anti-tumor drug camptothecin. With albumin serving as a nutrient to cells, the drug-encapsulated nanoparticle shows an enhanced ability to kill cancer cells compared to that of the free drug in solution both in vitro and in vivo.

Engineered polymer nanoparticles incorporating L-amino acid groups as affinity reagents for fibrinogen

Synthetic polymer hydrogel nanoparticles(NPs)were developed to function as abiotic affinity reagents for fibrinogen.These NPs were made using both temperature-sensitive N-isopropyl acrylamide(NIPAm)and L-amino acid monomers.Five kinds of L-amino acids were acryloylated to obtain functional monomers:L-phenylalanine(Phe)and L-leucine(Leu)with hydrophobic side chains,L-glutamic acid(Glu)with negative charges,and L-lysine(Lys)and L-arginine(Arg)with positive charges.After incubating the NPs with fibrinogen,g-globulin,and human serum albumin(HSA)respectively,the NPs that incorporated Nacryloyl-Arg monomers(AArg@NPs)showed the strongest and most specific binding affinity to fibrinogen,when compared with g-globulin and HSA.Additionally,the fibrinogen-AArg binding model had the best docking scores,and this may be due to the interaction of positively charged AArg@NPs and the negatively charged fibrinogen D domain and the hydrophobic interaction between them.The specific adsorption of AArg@NPs to fibrinogen was also confirmed by the immunoprecipitation assay,as the AArg@NPs selectively trapped the fibrinogen from a human plasma protein mixture.AArg@NPs had a strong selectivity for,and specificity to,fibrinogen and may be developed as a potential human fibrinogen-specific affinity reagent.