Immobilization of β-glucosidase on mercaptopropyl-functionalized mesoporous titanium dioxide

Mesoporous titanium dioxide (M-TiO₂) and mercaptopropyl-functionalized M-TiO₂ (SH-M-TiO₂) were prepared as carriers for immobilization of β-glucosidase (BG). Analyses suggested that in SH-M-TiO₂ the grafted mercaptopropyl groups attributed to about 3.5% of the total weight of the matrices. For the immobilization of BG, SH-M-TiO₂ or M-TiO₂ powder was added with free BG solution and stirred at 4 °C for 8 h, followed by centrifugation and washing. BG was effectively immobilized on SH-M-TiO₂ with an enzyme activity of 10.9 U/g, which was 92.8% of the total enzyme activity in the starting solution of free BG. BG immobilized on SH-M-TiO₂ (BG-SH-M-TiO₂) was more stable in pH, thermal, and storage tests than BG on M-TiO₂ or free BG in solution. The K_m and v_max of BG-SH-M-TiO₂ were similar to values for free BG in solution. Batch hydrolysis of cellobiose-containing substrate by BG-SH-M-TiO₂ was conducted. Conversion rates of cellobiose in 10 batches were consistently around 90%. Although the residual enzyme activity of BG-SH-M-TiO₂ decreased gradually, only a small amount of BG activity was detected in supernatants. This indicated effective adsorption of enzyme molecules to SH-M-TiO₂ matrices.     

一种新型网膜对SARS冠状病毒的抑制作用

利用光触媒钛羟基磷灰石网膜（PTAF）具有吸附和酶催化的特点,研究其对SARS病毒的抑制作用.实验结果表明在紫外照射条件下,PTAF膜对SARS冠状病毒的抑制率为100％.在没有紫外照射的条件下,PTAF膜对SARS冠状病毒的抑制率为99.99％, 与对照组相比,PTAF膜抑制病毒的效率是HAF膜的1 000倍以上.研究结果提示,PTAF在预防SARS冠状病毒及其他病毒性疾病流行方面有潜在的应用价值.     

Centerpiece微型钛板固定与传统丝线悬吊在颈椎后路单开门椎管扩大成形术中的应用效果比较研究

目的:比较Centerpiece微型钛板固定与传统丝线悬吊在颈椎后路单开门椎管扩大成形术中的应用效果。方法:选取于2015年2月～2017年9月期间北京大学第一医院收治的拟行颈椎后路单开门椎管扩大成形术的脊髓型颈椎病患者169例,根据治疗方式的不同将患者分为悬吊组(n=87,给予传统丝线悬吊治疗)和钛板组(n=82,给予Centerpiece微型钛板固定治疗),比较两组手术时间、术中出血量、术后颈椎疼痛时间、术后再关门发生率、轴性症状评分、日本骨科协会量表(JOA)评分、颈椎活动度、颈椎管矢状径、颈椎管横截面积、颈椎曲度、开门角度。结果:两组患者手术时间、术中出血量比较差异无统计学意义(P0.05),钛板组术后颈椎疼痛时间明显短于悬吊组,术后再关门发生率低于悬吊组(P0.05)。两组患者术后2个月、末次随访JOA评分均高于术前,且钛板组高于悬吊组(P0.05),两组患者末次随访轴性症状评分高于术后2个月,且钛板组术后2个月、末次随访轴性症状评分均高于悬吊组(P0.05)。与悬吊组相比,钛板组术后2个月、末次随访颈椎活动度、颈椎管矢状径升高(P0.05);而钛板组术后2个月颈椎管横截面积小于悬吊组,末次随访颈椎管横截面积大于悬吊组(P0.05)。悬吊组末次随访颈椎曲度小于术前、术后2个月(P0.05),钛板组手术前后颈椎曲度比较差异无统计学意义(P0.05);钛板组术后2个月、末次随访颈椎曲度、开门角度均大于悬吊组(P0.05)。结论:颈椎后路单开门椎管扩大成形术中应用Centerpiece微型钛板固定比传统丝线悬吊治疗的临床效果更好,可有效维持患者颈椎功能及活动度,并能改善神经功能。     

Overview of bacterial cellulose composites: A multipurpose advanced material

Bacterial cellulose (BC) has received substantial interest owing to its unique structural features and impressive physico-mechanical properties. BC has a variety of applications in biomedical fields, including use as biomaterial for artificial skin, artificial blood vessels, vascular grafts, scaffolds for tissue engineering, and wound dressing. However, pristine BC lacks certain properties, which limits its applications in various fields; therefore, synthesis of BC composites has been conducted to address these limitations. A variety of BC composite synthetic strategies have been developed based on the nature and relevant applications of the combined materials. BC composites are primarily synthesized through in situ addition of reinforcement materials to BC synthetic media or the ex situ penetration of such materials into BC microfibrils. Polymer blending and solution mixing are less frequently used synthetic approaches. BC composites have been synthesized using numerous materials ranging from organic polymers to inorganic nanoparticles. In medical fields, these composites are used for tissue regeneration, healing of deep wounds, enzyme immobilization, and synthesis of medical devices that could replace cardiovascular and other connective tissues. Various electrical products, including biosensors, biocatalysts, E-papers, display devices, electrical instruments, and optoelectronic devices, are prepared from BC composites with conductive materials. In this review, we compiled various synthetic approaches for BC composite synthesis, classes of BC composites, and applications of BC composites. This study will increase interest in BC composites and the development of new ideas in this field.     

Evaluation of four phosphopeptide enrichment strategies for mass spectrometry-based proteomic analysis

Information about phosphorylation status can be used to prioritize and characterize biological processes in the cell. Various analytical strategies have been proposed to address the complexity of phosphorylation status and comprehensively identify phosphopeptides. In this study, we evaluated four strategies for phosphopeptide enrichment, using titanium dioxide (TiO₂) and Phos-tag ligand particles from in-gel or in-solution digests prior to mass spectrometry-based analysis. Using TiO₂ and Phos-tag magnetic beads, it was possible to enrich phosphopeptides from in-gel digests of phosphorylated ovalbumin separated by Phos-tag SDS-PAGE or in-solution serum digests, while minimizing non-specific adsorption. The tip-column strategy with TiO₂ particles enabled enrichment of phosphopeptides from in-solution digests of whole-cell lysates with high efficiency and selectivity. However, the tip-column strategy with Phos-tag agarose beads yielded the greatest number of identified phosphopeptides. The strategies using both types of tip columns had a high degree of overlap, although there were differences in selectivity between the identified phosphopeptides. Together, our results indicate that multi-enrichment strategies using TiO₂ particles and Phos-tag agarose beads are useful for comprehensive phosphoproteomic analysis.     

Titanium oxide nanoparticles fabrication,hemoglobin interaction,white blood cells cytotoxicity,and antibacterial studies

This study is focused on the fabrication and characterization of titanium oxide (TiO₂) NPs. Afterwards; the interaction of TiO₂ NPs with human hemoglobin (Hb) was investigated by FTIR spectroscopy, fluorescence spectroscopy, and molecular docking studies. Also, the cytotoxic effect of fabricated TiO₂ NPs against human white blood cells (WBCs) was considered by MTT assay. The antibacterial effect of synthesized NPs was examined on Pseudomonas aeruginosa (ATCC 27853); Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923). TEM and DLS investigations showed that the synthesized TiO₂ NPs have a narrow nano-sized distribution. XRD pattern of the fabricated NPs exhibited that the TiO₂ NPs contain anatase phase. Similarity in amide I and II signal intensities showed that secondary structure of the adsorbed Hb is preserved. The intrinsic fluorescence study revealed that the fluorescence quenching of Hb was done by complex formation between Hb and TiO₂ NPs trough the hydrogen bond and van der Waals interactions. Synchronous fluorescence spectroscopy determined that interaction of TiO₂ NPs with Hb did not unfold the Hb structure in the vicinity of the Tyr and Trp residues. Molecular docking study depicted that Glu-95, Thr-134 and Tyr-140 are involved in the formation of hydrophilic bonds. MTT data and antibacterial assays indicated that TiO₂ NPs endow distinguished antibacterial activities against Gram-negative and Gram positive strains at safe concentrations. This study may reveal that fabricated TiO₂ NP can be used as a safe and potent antibacterial agent.

Communicated by Ramaswamy H. Sarma     

Identification of BoLA-DRB3.2 alleles in Korean native cattle (Hanwoo) and Holstein populations using a next generation sequencer

Bovine leucocyte antigen (encoded by BoLA) has been widely studied to identify the association with many traits related to immunity. Exon2 of BoLA-DRB3 is extremely polymorphic, and more than 100 alleles have been identified. We investigated polymorphisms of BoLA-DRB3.2 in Korean native cattle and Holstein populations using a next generation sequencer of the GS-FLX Titanium system. We found 38 alleles including 11 new alleles (BoLA-DRB3*1303, *4702, *7101, *7501, *7201, *7301, *7601, *1104, *7701, *7401 and *50021) in Hanwoo, and nine alleles including one new allele (BoLA-DRB3*7601) in Holstein. The 454 sequencing method is a promising alternative technology for high throughput genotyping of BoLA-DRB3.2 because of its technical advantages that allow it to overcome the disadvantages of sequence-based typing methods.     

Modified nano-TiO2 coupled with fluorescence spectroscopy for the separation/analysis of l-tryptophan

A novel adsorbent of carboxymethyl-β-cyclodextrin modified nanometer TiO₂ (CM-β-CD/TiO₂) was prepared and used as a solid-phase extraction (SPE) material coupled to fluorescence spectroscopy determination of l-tryptophan (l-Trp) in biological samples. The experimental conditions for modified nanometer TiO₂ separation/preconcentration of l-Trp were optimized. The adsorption capacity of CM-β-CD/TiO₂ for l-Trp was 75.2 μg/g. The linear range, detection limit (DL), and the relative standard deviation (RSD) were 0.10-1.20 μg/mL, 18.8 ng/mL, and 0.67% (n = 3, 1.0 μg/mL), respectively, with a preconcentration factor of 10. The developed method was applied to determination of l-Trp in real samples and the recoveries were found to be in the range of 99.2-100.3%. For validation, a comparison material of NIC-140686 sample was analyzed and the determined value was in good agreement with the certified value.     

Transition metals as protease inhibitors

An alternative approach to the development of clinically useful protease inhibitors was investigated. The approach utilized coordination chemistry of transition metal ions rather than substrate analogs to block active sites of these enzymes. In the case of serine proteases it was found that aqueous Ti(IV) is a potent inhibitor of the trypsin subclass, but not the chymotrypsin subclass. The direct binding of Ti(IV) to trypsin was made possible by the presence of a free carboxyl group at the bottom of the substrate binding pocket of the enzyme, and the five-coordinate geometry of TiO(SO₄)(H₂O). Although initial binding of Ti(IV) was reversible, it was followed in time by irreversible inhibition. Direct binding of octahedral or tetrahedral metal ion complexes was prevented by the inability of the enzyme active sites to promote formation of a five-coordinate transition state of the metal ion required for reaction. These studies demonstrate the ability of direct metal ion binding as a way to enhance blocking of enzyme active sites as compared with that of traditional organic inhibitors. Application of these findings was investigated by measuring the affect Ti(IV) had on growth ofEscherichia coli, Salmonella typhimurium, andPseudotnonas aeruginosa. Five-coordinate titanyl sulfate completely inhibited the growth of these organisms. This suggests that five-coordinate titanyl sulfate, which is easier and less expensive to manufacture than conventional antibiotics, may be useful in controlling endemic infections ofE. coli andS. typhimurium.