Design,synthesis, and evaluation of guanylhydrazones as potential inhibitors or reactivators of acetylcholinesterase

Analogs of pralidoxime, which is a commercial antidote for intoxication from neurotoxic organophosphorus compounds, were designed, synthesized, characterized, and tested as potential inhibitors or reactivators of acetylcholinesterase (AChE) using the Ellman’s test, nuclear magnetic resonance, and molecular modeling. These analogs include 1-methylpyridine-2-carboxaldehyde hydrazone, 1-methylpyridine-2-carboxaldehyde guanylhydrazone, and six other guanylhydrazones obtained from different benzaldehydes. The results indicate that all compounds are weak AChE reactivators but relatively good AChE inhibitors. The most effective AChE inhibitor discovered was the guanylhydrazone derived from 2,4-dinitrobenzaldehyde and was compared with tacrine, displaying similar activity to this reference material. These results indicate that guanylhydrazones as well as future similar derivatives may function as drugs for the treatment of Alzheimer's disease.     

Bioluminescence‐based assays for assessing eco‐ and genotoxicity of airborne emissions

Ecotoxicological tests (or bioassays) are controlled, reproducible tests where ecological responses are determined quantitatively. Due to numerous difficulties arising when airborne emissions are sampled, relatively few ecotoxicological assays have been applied. Aerosol particles are generally collected on a filter, which limits the quantity of the sample, thus also limiting the range of available test organisms. Bacterial bioassays require low sample quantity, and make a good choice when eco‐ or genotoxicity of the sample are to be determined. Of bacterial assays, the bioluminescence inhibition test has been proven applicable for assessing toxicity of airborne contaminants. In this paper diverse test protocols and their modifications are reviewed. Copyright © 2016 John Wiley & Sons, Ltd.     

Similar recent selection criteria associated with different behavioural effects in two dog breeds

Selection during the last decades has split some established dog breeds into morphologically and behaviourally divergent types. These breed splits are interesting models for behaviour genetics since selection has often been for few and well‐defined behavioural traits. The aim of this study was to explore behavioural differences between selection lines in golden and Labrador retriever, in both of which a split between a common type (pet and conformation) and a field type (hunting) has occurred. We hypothesized that the behavioural profiles of the types would be similar in both breeds. Pedigree data and results from a standardized behavioural test from 902 goldens (698 common and 204 field) and 1672 Labradors (1023 and 649) were analysed. Principal component analysis revealed six behavioural components: curiosity, play interest, chase proneness, social curiosity, social greeting and threat display. Breed and type affected all components, but interestingly there was an interaction between breed and type for most components. For example, in Labradors the common type had higher curiosity than the field type (F_1,1668 = 18.359; P < 0.001), while the opposite was found in goldens (F_1,897 = 65.201; P < 0.001). Heritability estimates showed considerable genetic contributions to the behavioural variations in both breeds, but different heritabilities between the types within breeds was also found, suggesting different selection pressures. In conclusion, in spite of similar genetic origin and similar recent selection criteria, types behave differently in the breeds. This suggests that the genetic architecture related to behaviour differs between the breeds.     

The influence of AVPR1A genotype on individual differences in behaviors during a mirror self‐recognition task in chimpanzees (Pan troglodytes)

The mark/rouge test has been used to assess mirror self‐recognition (MSR) in many species. Despite consistent evidence of MSR in great apes, genetic or non‐genetic factors may account for the individual differences in behavioral responses that have been reported. We examined whether vasopressin receptor gene (AVPR1A) polymorphisms are associated with MSR‐related behaviors in chimpanzees since vasopressin has been implicated in the development and evolution of complex social relations and cognition and chimpanzees are polymorphic for the presence of the RS3‐containing DupB region. We compared a sample of DupB+/? and DupB?/? chimpanzees on a mark test to assess its role on social behavior toward a mirror. Chimpanzees were administered two, 10‐min sessions where frequencies of mirror‐guided self‐directed behaviors, contingent actions and other social behaviors were recorded. Approximately one‐third showed evidence of MSR and these individuals exhibited more mirror‐guided self‐exploratory behaviors and mouth contingent actions than chimpanzees not classified as passers. Moreover, DupB+/? males exhibited more scratching and agonistic behaviors than other male and female cohorts. Our findings support previous studies demonstrating individual differences in MSR abilities in chimpanzees and suggest that AVPR1A partly explains individual differences in MSR by influencing the behavioral reactions of chimpanzees in front of a mirror.     

Effect of lipid composition on incorporation of trastuzumab-PEG-lipid into nanoliposomes by post-insertion method: physicochemical and cellular characterization

Context: Anti-HER2 immunoliposomes are promising nanotechnology based systems for active targeting of breast tumors, which depends on the amount of incorporated antibody.

Objective/Aim: In this work, we investigated the possible effect of lipid composition on the incorporation of trastuzumab-PEG-PE micelles into nanoliposomes and on their subsequent specific cellular targeting.

Materials and methods: Trastuzumab (anti-HER2 monoclonal antibody) was monothiolated and conjugated to maleimide-PEG-PE micelles. Liposomes of different lipid compositions were prepared by the thin layer hydration. Trastuzumab-PEG-PE micelles were incorporated into the liposomes by the post-insertion method. The percentage of lipid mixing was determined based on fluorescence resonance energy transfer. Cellular binding and uptake of rhodamine-labeled immunoliposomes were studied in SKBR-3 (HER2⁺⁺⁺) and MCF-7 (HER2⁺) cells. Also, antitumor cell activity of the immunoliposomes was compared to free trastuzumab and the liposomes.

Results: The lipid mixing of trastuzumab-PEG-PE micelles depended on the liposome composition. The immunoliposomes containing DPPC, cholesterol and PEG-PE showed prominent lipid mixing. The lipid mixing was consistent with the cell binding results which showed an efficient and specific binding of the immunoliposomes to SKBR-3 cells. Antitumor cell activity of the immunoliposomes in SKBR-3, unlike MCF-7 cells, depended on the content of trastuzumab.

Discussion: Cholesterol and PEG-PE in the liposome composition are prerequisites for a successful lipid mixing due to their ability to facilitate fusion. The higher lipid mixing results in higher antibody incorporation and consequently higher targeted cell binding.

Conclusions: The lipid mixing depends on the liposome composition, which reflects targeted cell binding of the immunoliposomes.     

胡椒木叶片精油成分分析及其抗氧化、驱虫、抗菌活性

为解决在城市绿化中滥用化学农药引起的生态问题,研究了胡椒木(Zanthoxylum piperitum)叶片的精油成分及其驱虫、杀虫、杀菌活性。结果表明,采用石油醚萃取胡椒木叶片精油的最佳工艺条件为提取时间4 h、温度50℃、料液比1:5,得率最高为0.5123%。从胡椒木叶片精油中共鉴定出25个化学成分,主要为酯类、烃类和醇类,占总峰面积的99.96%,而具有驱虫、杀虫和抗菌活性的成分占88.50%,包括肉桂酸甲酯(87.83%)、柠檬烯(0.18%)、薄荷醇(0.11%)、香茅醛(0.10%)、α-蒎烯(0.08%)、α-石竹烯(0.08%)、松油醇(0.04%)、石竹烯(0.03%)、羟基大牻牛儿-1(10),5-二烯(0.03%)和β-水芹烯(0.02%)等。20 mg m L~(–1)的精油对DPPH·自由基的清除率达85.38%,接近BHT阳性对照;但还原性低于Vc,表现出较弱活性,这可能与提取工艺有关。因此,胡椒木叶精油含有多种驱虫、杀虫、抗菌等生物活性成分,具有开发为天然植物农药的潜力。     

细菌VBNC态检测技术的研究进展

细菌在一些不良压力条件下会进入"活的非可培养状态"(viable but non-culturable,VBNC),其仍具有活力但不能采用常规的平板培养基进行培养,VBNC态细菌一旦培养条件适宜仍能继续生长繁殖,有些致病菌依旧具有毒力。如果检测方法不当,会造成假阳性或假阴性的结果,需要采取适合的检测方法进行检测。文章概述了几种检测VBNC态细菌的方法,如活菌直接计数法、核酸染料检测法、呼吸检测法、分子生物学法、免疫学法、流式细胞仪检测法等,并对检测方法的应用现状进行了评述。     

New hybrid molecules with anticonvulsant and antinociceptive activity derived from 3-methyl- or 3,3-dimethyl-1-[1-oxo-1-(4-phenylpiperazin-1-yl)propan-2-yl]pyrrolidine-2,5-diones

The purpose of this study was to synthetize the focused library of 34 new piperazinamides of 3-methyl- and 3,3-dimethyl-(2,5-dioxopyrrolidin-1-yl)propanoic or butanoic acids as potential new hybrid anticonvulsants. These hybrid molecules join the chemical fragments of well-known antiepileptic drugs (AEDs) such as ethosuximide, levetiracetam, and lacosamide. Compounds 5–38 were prepared in a coupling reaction of the 3-methyl- or 3,3-dimethyl-2-(2,5-dioxopyrrolidin-1-yl)propanoic (1, 2) or butanoic acids (3, 4) with the appropriately substituted secondary amines in the presence of the N,N-carbonyldiimidazole reagent. The initial anticonvulsant screening was performed in mice (ip) using the ‘classical’ maximal electroshock (MES) and subcutaneous pentylenetetrazole (scPTZ) tests as well as in the six-Hertz (6 Hz) model of pharmacoresistant limbic seizures. The acute neurological toxicity was determined applying the chimney test. The broad spectra of activity across the preclinical seizure models in mice ip displayed compounds 7, 15, and 36. The most favorable anticonvulsant properties demonstrated 15 (ED₅₀ MES = 74.8 mg/kg, ED₅₀ scPTZ = 51.6 mg/kg, ED₅₀ 6 Hz = 16.8 mg/kg) which showed TD₅₀ = 213.3 mg/kg in the chimney test that yielded satisfying protective indexes (PI MES = 2.85, PI scPTZ = 4.13, PI 6 Hz = 12.70) at time point of 0.5 h. As a result, compound 15 displayed comparable or better safety profile than clinically relevant AEDs: ethosuximide, lacosamide or valproic acid. In the in vitro assays compound 15 was observed as relatively effective binder to the neuronal voltage-sensitive sodium and L-type calcium channels. Beyond the anticonvulsant properties, 6 compounds diminished the pain responses in the formalin model of tonic pain in mice.     

Mechanical properties of mosquito nets in the context of hernia repair

The paper deals with issue of applying mosquito nets as implants in hernia repair, which have already been used in resource-poor developing countries. Uniaxial tensile tests have been conducted on polyester mosquito meshes in two orthogonal directions. Non-linear elastic constitutive laws parameters have been identified to be applied in dense net material models. Mechanical performance of tested mosquito nets has been compared with properties of commercial implants used in treatment of hernia and with properties of human tissue. This study contributes to mechanical knowledge of hernia repair issue by investigation of cheaper alternative to commercial implants.     

Patients with chronic,but not episodic,migraine display altered activity of their neck extensor muscles

The current study aimed to investigate differences in activity of neck flexor and extensor muscles in women with migraine considering the chronicity of their condition. Thirty-one subjects with episodic migraine, 21 with chronic migraine and 31 healthy controls participated. Surface electromyography signals were recorded bilaterally from the sternocleidomastoid, anterior scalene, splenius capitis and upper trapezius muscles as subjects performed 5 stages of cranio-cervical flexion (CCF), representing a progressive increase in range of CCF motion. Comparison of normalized root-mean-square among groups was conducted with 3 × 5 ANCOVA with task level as the within-subject variable, group as the between-subject variable, and the presence of neck pain and disability as co-variates. The group with chronic migraine exhibited increased activity of their extensor muscles compared to the control and episodic migraine groups (splenius capitis: F = 3.149, P = 0.045; upper trapezius: F = 3.369, P = 0.041). No significant between-group differences were found for the superficial neck flexors (sternocleidomastoid: F = 1.161, P = 0.320; anterior scalene: F = 0.135, P = 0.874). In conclusion, women with chronic migraine exhibit increased activity of their superficial neck extensor muscles when acting as antagonists during low-load isometric CCF contractions in comparison to non-headache subjects.