Purification and characterization of an antimicrobial protein from Gastrodia elata Blume tubers

Purpose: To purify and characterize a novel antimicrobial protein from the Gastrodia elata Blume (Bl.) plant, which has long been used in herbal medicine.Methods: The procedure for isolation and purification of Gastrodia elata protein (GEP) involved phosphate buffer extraction, ammonium sulfate precipitation, ion-exchange chromatography, and gelfiltration chromatography. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis was employed to detect the apparent molecular mass and determine homogeneity, while paper disc diffusion was used to measure the antibacterial activity of GEP. A hemolytic assay was performed on rabbit red blood cells. The effect of pH, salt concentration, and temperature on the antibacterial activity of GEP was evaluated by minimum inhibitory concentration assay.Results: GEP was a 14-kDa monomer and displayed antimicrobial activity against Staphylococcus aureus and Candida albicans, with 8.0-mm and 9.4-mm zones of inhibition, respectively, but no antibacterial activity was observed against Escherichia coli. GEP had little hemolytic activity on red blood cells even at a concentrations of up to 200 mg/ml. GEP was thermally stable at temperatures below 70 °C for 30 min, and displayed higher antibacterial activity in the pH range 5.0 to 7.0.Conclusion: GEP protein is relatively thermostable and possesses antimicrobial activity. The results suggest that GEP protein has potential agricultural and industrial applications, such as in transgenic plants.Keywords: Antimicrobial protein, Gastrodia elata, Protein characterizatio

New tumor-targeted nanosized delivery carrier for oligonucleotides: characteristics in vitro and in vivo

Tianyang Zhou1,2, Xin Jia1, Huixiang Li3, Jin Wang3, Hongling Zhang1,2, Youmei A1,2, Zhenzhong Zhang1,21School of Pharmaceutical Sciences, 2Nanotechnology Research Center for Drugs, 3Department of Pathology, Medical School of Zhengzhou University, Zhengzhou, People’s Republic of ChinaBackground: The purpose of this study was to investigate the in vitro and in vivo characteristics of a new tumor-targeted nanosized delivery carrier for antisense oligonucleotide (ASON).Methods: Polyethylenimine (PEI) was used to condense ASON to form nanosized complexes (PEI/ASON), which were then modified using asparagine-glycine-arginine (NGR) peptide to obtain a tumor-targeted nanosized delivery carrier (NGR/PEI/ASON). The conditions required to form PEI/ASON were investigated.Results: A linear correlation between the natural logarithm of the N/P ratio (PEI to ASON) and the zeta potential of the PEI/ASON complexes was found, ranging from 1.5 to 5.0. The pH of the solution strongly influenced the zeta potential of the PEI/ASON complexes. PEI/ASON and NGR/PEI/ASON were stable in RPMI-1640 culture medium in the presence of Dextran 70. Incorporation of ASON into PEI/ASON and NGR/PEI/ASON complexes prevented degradation of ASON by DNase I.Conclusion: Both ASON/PEI and NGR/PEI/ASON complexes enhanced the uptake of ASON by EC9706 cells in vitro. In vivo, NGR/PEI/ASON complexes had the ability to target tumor tissues effectively.Keywords: nanosized delivery system, squamous cell carcinoma, antisense oligonucleotid

Bounded real lemmas for positive descriptor systems

A well known result in the theory of linear positive systems is the existence of positive definite diagonal matrix (PDDM) solutions to some well known linear matrix inequalities (LMIs). In this paper, based on the positivity characterization, a novel bounded real lemma for continuous positive descriptor systems in terms of strict LMI is first established by the separating hyperplane theorem. The result developed here provides a necessary and sufficient condition for systems to possess H?H? norm less than ? and shows the existence of PDDM solution. Moreover, under certain condition, a simple model reduction method is introduced, which can preserve positivity, stability and H?H? norm of the original systems. An advantage of such method is that systems? matrices of the reduced order systems do not involve solving of LMIs conditions. Then, the obtained results are extended to discrete case. Finally, a numerical example is given to illustrate the effectiveness of the obtained results

AcoMYB4, an Ananas comosus L. MYB transcription factor, functions in osmotic stress through negative regulation of ABA signaling

Drought and salt stress are the main environmental cues affecting the survival, development, distribution, and yield of crops worldwide. MYB transcription factors play a crucial role in plants’ biological processes, but the function of pineapple MYB genes is still obscure. In this study, one of the pineapple MYB transcription factors, AcoMYB4, was isolated and characterized. The results showed that AcoMYB4 is localized in the cell nucleus, and its expression is induced by low temperature, drought, salt stress, and hormonal stimulation, especially by abscisic acid (ABA). Overexpression of AcoMYB4 in rice and Arabidopsis enhanced plant sensitivity to osmotic stress; it led to an increase in the number stomata on leaf surfaces and lower germination rate under salt and drought stress. Furthermore, in AcoMYB4 OE lines, the membrane oxidation index, free proline, and soluble sugar contents were decreased. In contrast, electrolyte leakage and malondialdehyde (MDA) content increased significantly due to membrane injury, indicating higher sensitivity to drought and salinity stresses. Besides the above, both the expression level and activities of several antioxidant enzymes were decreased, indicating lower antioxidant activity in AcoMYB4 transgenic plants. Moreover, under osmotic stress, overexpression of AcoMYB4 inhibited ABA biosynthesis through a decrease in the transcription of genes responsible for ABA synthesis (ABA1 and ABA2) and ABA signal transduction factor ABI5. These results suggest that AcoMYB4 negatively regulates osmotic stress by attenuating cellular ABA biosynthesis and signal transduction pathways

Estimates on compressed neural networks regression

When the neural element number nn of neural networks is larger than the sample size mm, the overfitting problem arises since there are more parameters than actual data (more variable than constraints). In order to overcome the overfitting problem, we propose to reduce the number of neural elements by using compressed projection AA which does not need to satisfy the condition of Restricted Isometric Property (RIP). By applying probability inequalities and approximation properties of the feedforward neural networks (FNNs), we prove that solving the FNNs regression learning algorithm in the compressed domain instead of the original domain reduces the sample error at the price of an increased (but controlled) approximation error, where the covering number theory is used to estimate the excess error, and an upper bound of the excess error is given

Personalized antiplatelet therapy guided by clopidogrel pharmacogenomics in acute ischemic stroke and transient ischemic attack: A prospective, randomized controlled trial

Background: Clopidogrel is frequently used in patients with ischemic stroke or transient ischemic attack (TIA), but its efficacy is hampered by inter-individual variability, due to genetic differences associated with clopidogrel metabolism. We conducted this randomized controlled trial to validate whether the personalized antiplatelet therapy based on clopidogrel pharmacogenomics and clinical characteristics leads to better clinical outcomes compared with standard treatment.Methods: Patients were randomly divided into the standard group or pharmacogenetic group, in which the pharmacogenetic group required the detection of the genotyping of CYP2C19*2, CYP2C19*3, and CYP2C19*17. Patients were followed up for 90 days for the primary efficacy endpoint of new stroke events, secondary efficacy endpoint of individual or composite outcomes of the new clinical vascular events, and the incidence of disability. The primary safety outcome was major bleeding.Results: A total of 650 patients underwent randomization, among which 325 were in the pharmacogenomics group while 325 were in the standard group. Our study found after a 90-day follow-up, the risk of stroke and composite vascular events in the pharmacogenomics group was lower than that in the standard group. The incidence of disability significantly decreased in the pharmacogenomics group. In addition, no statistically significant differences were observed in bleeding events between the two groups.Conclusion: The present study demonstrates that personalized antiplatelet therapy guided by clopidogrel pharmacogenomics and clinical characteristics can significantly improve the net clinical benefit of ischemic stroke or TIA patients during the 90-day treatment period without increasing bleeding risk

Small Molecule, Non-Peptide p75NTR Ligands Inhibit Aβ-Induced Neurodegeneration and Synaptic Impairment

The p75 neurotrophin receptor (p75NTR) is expressed by neurons particularly vulnerable in Alzheimer's disease (AD). We tested the hypothesis that non-peptide, small molecule p75NTR ligands found to promote survival signaling might prevent Aβ-induced degeneration and synaptic dysfunction. These ligands inhibited Aβ-induced neuritic dystrophy, death of cultured neurons and Aβ-induced death of pyramidal neurons in hippocampal slice cultures. Moreover, ligands inhibited Aβ-induced activation of molecules involved in AD pathology including calpain/cdk5, GSK3β and c-Jun, and tau phosphorylation, and prevented Aβ-induced inactivation of AKT and CREB. Finally, a p75NTR ligand blocked Aβ-induced hippocampal LTP impairment. These studies support an extensive intersection between p75NTR signaling and Aβ pathogenic mechanisms, and introduce a class of specific small molecule ligands with the unique ability to block multiple fundamental AD-related signaling pathways, reverse synaptic impairment and inhibit Aβ-induced neuronal dystrophy and death