APPLICABILITY OF CAPILLARY ELECTROPHORESIS IN PROTEIN SEPARATIONS WITH REGARD TO WHEAT AND WHEAT ANALOGOUS PROTEINS

High-performance capillary electrophoresis (HPCE), among other analytical techniques (e.g. acid polyacrylamide gel electrophoresis, sodium dodecyl sulphate polyacrylamide gel electrophoresis and reversed-phase high-performance liquid chromatography) [1]-[4] is more and more widely used also in the field of separation of cereal proteins [5]-[18]. HPCE is versatile, easy to automate, does not need toxic reagents or long analysis time, but requires only small sample size, small amount of buffer and provides high-resolution separations. So this analytical technique is particularly applicable for studying the fine structure of the composition of wheat proteins. Capillary electrophoresis has been used at our department for the determination of the fine structure of different wheat protein fractions [19]. Differences between the various wheat cultivars and changes during grain maturation process have also been studied using a home-built capillary electrophoretic system [20]. The same system has been used to investigate the electrophoretic properties of a gliadin analogous protein (BM180 - a basement membrane protein with a potential autoantigen role) [21]. The more strictly controllable nature of an automated system (especially the temperature control of the capillary) allows the use of higher voltages, so separation time decreases and resolution increases. Purchasing an automated capillary electrophoretic system we gained an opportunity to compare the two electrophoretic systems also in the field of wheat protein analyses. The aim of present work was to give an impression of the work done by capillary electrophoresis at our department

Proof-of-Concept for the Analgesic Effect and Thermoregulatory Safety of Orally Administered Multi-Target Compound SZV 1287 in Mice: A Novel Drug Candidate for Neuropathic Pain

SZV 1287 (3-(4,5-diphenyl-1,3-oxazol-2-yl)propanal oxime) is a novel multi-target candidate under preclinical development for neuropathic pain. It inhibits amine oxidase copper containing 3, transient receptor potential ankyrin 1 and vanilloid 1 (TRPV1) receptors. Mainly under acidic conditions, it is transformed to the cyclooxygenase inhibitor oxaprozin, which is ineffective for neuropathy. Therefore, an enterosolvent capsule is suggested for oral formulation, which we investigated for nociception, basic kinetics, and thermoregulatory safety in mice. The antihyperalgesic effect of SZV 1287 (10, 20, 50, and 200 mg/kg, p.o.) was determined in partial sciatic nerve ligation-induced traumatic neuropathy by aesthesiometry, brain and plasma concentrations by HPLC, and deep body temperature by thermometry. Its effect on proton-induced TRPV1 activation involved in thermoregulation was assessed by microfluorimetry in cultured trigeminal neurons. The three higher SZV 1287 doses significantly, but not dose-dependently, reduced neuropathic hyperalgesia by 50% of its maximal effect. It was quickly absorbed; plasma concentration was stable for 2 h, and it entered into the brain. Although SZV 1287 significantly decreased the proton-induced TRPV1-mediated calcium-influx potentially leading to hyperthermia, it did not alter deep body temperature. Oral SZV 1287 inhibited neuropathic hyperalgesia and, despite TRPV1 antagonistic action and brain penetration, it did not influence thermoregulation, which makes it a promising analgesic candidate

A simple modification increases specificity and efficiency of asymmetric PCR

Although various methods have been developed to suffice the oligonucleotide demand of molecular biology laboratories, in vitro production of high-purity ssDNAs remains to be a challenging task. We hypothesized that complementing the asymmetric PCR with 3’ phosphate blocked limiting primer decreases the mispriming thus reduces polymerisation of DNA by-products. The presented results attest our assumption that the primer blocked asymmetric PCR (PBA-PCR) selectively produces ssDNA of interest and is even suitable for effective amplification of DNA libraries of large sequence space. The high-throughput sequence analysis demonstrated that PBA-PCR also alleviates the PCR bias obstacle since it does not distort the sequence space. The practicability of the novel method was verified by monitoring the process of SELEX and screening of aptamer candidates using PBA-PCR produced ssDNAs in Amplified Luminescent Proximity Homogeneous Assay. In summary, we have developed a generally applicable method for straightforward, cost-effective production of ssDNA with on demand labelling. © 2018 Elsevier B.V