Sonication assisted assemblage of exotic polymer supported nanostructured bio-hybrid system and prospective application

This work was focused on sonication mediated immobilization of porcine pancreatic lipase (PPL) onto poly(ethylene glycol) supported silver-iron oxide hybrid nanoparticles (PEG-Ag/IONPs). Selected process parameters of sonication were optimized using response surface methodology. Sonication assisted assemblage of spherical PEG-Ag/IONPs and consequent evolution of nanorods post PPL immobilization were documented. The efficacy of the reported immobilization strategy was attested by the increased thermostability, storage stability and enhanced activity of the biocatalyst, suggestive of plausible structural modulations post immobilization. The commercial prospect of the antibacterial and magnetically recyclable system was vouched by its excellent compatibility with some commercial detergents for oil de-staining.     

Identification of Phenolic Compounds by LC-MS/MS and Evaluation of Bioactive Properties of Two Edible Halophytes: Limonium effusum and L. sinuatum

This work aimed to evaluate the phenolic content and in vitro antioxidant, antimicrobial and enzyme inhibitory activities of the methanol extracts and their fractions of two edible halophytic Limonium species, L. effusum (LE) and L. sinuatum (LS). The total phenolic content resulted about two-fold higher in the ethyl acetate fraction of LE (522.82 ± 5.67 mg GAE/g extract) than in that of LS (274.87 ± 1.87 mg GAE/g extract). LC-MS/MS analysis indicated that tannic acid was the most abundant phenolic acid in both species (71,439.56 ± 3643.3 µg/g extract in LE and 105,453.5 ± 5328.1 µg/g extract in LS), whereas hyperoside was the most abundant flavonoid (14,006.90 ± 686.1 µg/g extract in LE and 1708.51 ± 83.6 µg/g extract in LS). The antioxidant capacity was evaluated by DPPH and TAC assays, and the stronger antioxidant activity in ethyl acetate fractions was highlighted. Both species were more active against Gram-positive bacteria than Gram negatives and showed considerable growth inhibitions against tested fungi. Interestingly, selective acetylcholinesterase (AChE) activity was observed with LE and LS. Particularly, the water fraction of LS strongly inhibited AChE (IC₅₀ = 0.199 ± 0.009 µg/mL). The ethyl acetate fractions of LE and LS, as well as the n-hexane fraction of LE, exhibited significant antityrosinase activity (IC₅₀ = 245.56 ± 3.6, 295.18 ± 10.57 and 148.27 ± 3.33 µg/mL, respectively). The ethyl acetate fraction and methanol extract of LS also significantly inhibited pancreatic lipase (IC₅₀ = 83.76 ± 4.19 and 162.2 ± 7.29 µg/mL, respectively). Taken together, these findings warrant further investigations to assess the potential of LE and LS as a bioactive source that can be exploited in pharmaceutical, cosmetics and food industries.     

TTF-1、SPA 表达与Ⅱ期肺腺癌患者术后复发及生存关系的研究

目的 探讨甲状腺转录因子-1（TTF-1）、肺表面活性物质A（SPA）的表达与Ⅱ期肺腺癌患者术后复发及生存的相关 性。方法 收集经根治手术治疗的104 例Ⅱ期原发性肺腺癌患者资料,采用免疫组化法综合染色强度和阳性细胞所占比例进行半定量测定肿瘤组织的TTF-1、SPA 表达,分析两者与患者术后复发及生存的相关性。结果 TTF-1 阳性表达患者术后2 年复发率较阴性表达患者低[61.2%（60/98）vs 83.3%（5/6）],术后生存期较长[（26.00±1.89）个月vs（22.00±8.76）个月],但差异均无统计学意义（均P＞0.05）。SPA 阳性表达患者术后2年复发率较阴性表达患者明显为低[54.8%（34/62）vs 79.2%（19/24）],术后生存期明显较长[（30.00±1.41）个月vs（21.00±2.18）个月],差异均有统计学意义（均P＜0.05）。结论 SPA 阳性表达预示Ⅱ期原发性肺腺癌患者术后2年复发率低,生存期长。TTF-1 表达与Ⅱ期肺腺癌患者术后复发及生存有一定的相关性。     

A G-Quadruplex-Binding Small Molecule and the HDAC Inhibitor SAHA (Vorinostat) Act Synergistically in Gemcitabine-Sensitive and Resistant Pancreatic Cancer Cells

The stabilisation of G-quadruplexes (G4s) by small-molecule compounds is an effective approach for causing cell growth arrest, followed by cell death. Some of these compounds are currently being developed for the treatment of human cancers. We have previously developed a substituted naphthalene diimide G4-binding molecule (CM03) with selective potency for pancreatic cancer cells, including gemcitabine-resistant cells. We report here that CM03 and the histone deacetylase (HDAC) inhibitor SAHA (suberanilohydroxamic acid) have synergistic effects at concentrations close to and below their individual GI₅₀ values, in both gemcitabine-sensitive and resistant pancreatic cancer cell lines. Immunoblot analysis showed elevated levels of γ-H2AX and cleaved PARP proteins upon drug combination treatment, indicating increased levels of DNA damage (double-strand break events: DSBs) and apoptosis induction, respectively. We propose that the mechanism of synergy involves SAHA relaxing condensed chromatin, resulting in higher levels of G4 formation. In turn, CM03 can stabilise a greater number of G4s, leading to the downregulation of more G4-containing genes as well as a higher incidence of DSBs due to torsional strain on DNA and chromatin structure.     

Effective Tumor Targeting by EphA2-Agonist-Biotin-Streptavidin Conjugates

We recently reported on a potent synthetic agent, 135H11, that selectively targets the receptor tyrosine kinase, EphA2. While 135H11 possesses a relatively high binding affinity for the ligand-binding domain of EphA2 (Kd~130 nM), receptor activation in the cell required the synthesis of dimeric versions of such agent (namely 135H12). This was expected given that the natural ephrin ligands also need to be dimerized or clustered to elicit agonistic activity in cell. In the present report we investigated whether the agonistic activity of 135H11 could be enhanced by biotin conjugation followed by complex formation with streptavidin. Therefore, we measured the agonistic EphA2 activity of 135H11-biotin (147B5) at various agent/streptavidin ratios, side by side with 135H12, and a scrambled version of 147B5 in pancreatic- and breast-cancer cell lines. The (147B5)_n-streptavidin complexes (when n = 2, 3, 4, but not when n = 1) induced a strong receptor degradation effect in both cell lines compared to 135H12 or the (scrambled-147B5)₄-streptavidin complex as a control, indicating that multimerization of the targeting agent resulted in an increased ability to cause receptor clustering and internalization. Subsequently, we prepared an Alexa-Fluor-streptavidin conjugate to demonstrate that (147B5)₄-AF-streptavidin, but not the scrambled equivalent complex, concentrates in pancreatic and breast cancers in orthotopic nude-mouse models. Hence, we conclude that these novel targeting agents, with proper derivatization with imaging reagents or chemotherapy, can be used as diagnostics, and/or to deliver chemotherapy selectively to EphA2-expressing tumors.     

N-Acetyl peak in MR spectra of intracranial metastatic mucinous adenocarcinomas

Absence of N-acetylaspartate (NAA) is one important diagnostic criterion of MR spectroscopy (MRS) that may suggest that an intracranial mass lesion is a metastasis. We report two cases of histopathology-confirmed intracranial metastatic mucinous adenocarcinoma, which predominantly showed a large metabolite peak at 2.0 ppm, mimicking an NAA peak of normal brain tissue. This finding could be of help in the interpretation of MRS in cases of intracranial enhancing mass lesions, metastases or gliomas.     

Determination of aprotinin by high-performance liquid chromatography

Summary A highly accurate and reproducible method for the determination of aprotinin (bovine pancreatic trypsin inhibitor) by HPLC is described. In the experiments, the relative standard deviation was 1.2% and detection limit 1 FIP-U cm^–3. Also, the method is quick and selective and active ingredients from difference source correlate well with enzymatic method. Analyses at different laboratories can be compared directly.     

Preparation and characterization of pancreatic lipase immobilized in Eudragit-matrix

Pancreatic lipase (EC 3.1.1.3) was immobilized by entrapping in a commercial preparation of acrylic/methacrylic acid ester-based copolymer (Eudragit E 30 D). The activity of the immobilized lipase beads with a diameter of 1.5-2.0 mm was found to be lower than that of the free lipase. The optimum pH was shifted to the alkaline region and the thermal stability increased, whereas the optimum temperature level remained unchanged. The most important reason for the decreased activity was diffusion limitations. The diffusion of the substrate and products became more pronounced, and lipolytic activity increased upon addition of n-hexane into the reaction medium. The storage and operational stabilities of the immobilized lipase were investigated, and both characteristics were found to be increased when compared to the free enzyme. Furthermore, mechanical or magnetic stirring during the operation were found to have no influence on the carrier-matrix as determined by nephelometric measurements.