Liberation and oxygenation of polyenoic acids in stimulated platelets

Radiolabeled polyenoic acids were incorporated into human platelet lipids using albumin as vector. Platelets were then triggered with 0.1 or 1 U/ml thrombin, and 0.5 or 2 x 10(-6) M calcium ionophore A23187. Lipid extracts were analyzed for neutral lipids, free fatty acids, monohydroxylated acids, prostanoids and glycocerophospholipid subclasses. During platelet activation induced by thrombin or by ionophore, arachidonic and eicosapentaenoic acids were liberated from phospholipids in large amounts and were subsequently oxygenated via platelet oxygenases. Substantial amounts of lipoxygenase products and thromboxanes were produced from these acids. Liberation and oxygenation of linoleic, alpha-linolenic, and docosahexaenoic acids were much less pronounced. Polyenoic acid liberation from phospholipid subclasses also behaved quite differently. Apart from alpha-linolenic and adrenic acids, which were poorly liberated, all the others were freed from phosphatidylinositol. In addition, arachidonic, eicosapentaenoic, and 5, 8, 11-eicosatrienoic acids were liberated from phosphatidylcholine at high concentrations of agonists and partially reincorporated into phosphatidylethanolamine. Finally, linoleic acid was deacylated from phosphatidylinositol and phosphatidylserine and almost entirely reacylated into phosphatidylcholine, whereas docosahexaenoic acid was deacylated from phosphatidylcholine and phosphatidylinositol reacylated into phosphatidylethanolamine, respectively. It is concluded that these polyenoic acids, all for which modulate platelet functions, exhibit very different metabolisms. They may act via their oxygenated derivatives and/or at the membrane phospholipid level.     

First report of computational protein–ligand docking to evaluate susceptibility to HIV integrase inhibitors in HIV-infected Iranian patients

BackgroundIran has recently included integrase (INT) inhibitors (INTIs) in the first‐line treatment regimen in human immunodeficiency virus (HIV)-infected patients. However, there is no bioinformatics data to elaborate the impact of resistance-associated mutations (RAMs) and naturally occurring polymorphisms (NOPs) on INTIs treatment outcome in Iranian patients.MethodIn this cross-sectional survey, 850 HIV-1-infected patients enrolled; of them, 78 samples had successful sequencing results for INT gene. Several analyses were performed including docking screening, genotypic resistance, secondary/tertiary structures, post-translational modification (PTM), immune epitopes, etc.ResultThe average docking energy (E value) of different samples with elvitegravir (EVG) and raltegravir (RAL) was more than other INTIs. Phylogenetic tree analysis and Stanford HIV Subtyping program revealed HIV-1 CRF35-AD was the predominant subtype (94.9%) in our cases; in any event, online subtyping tools confirmed A1 as the most frequent subtype. For the first time, CRF-01B and BF were identified as new subtypes in Iran. Decreased CD4 count was associated with several factors: poor or unstable adherence, naïve treatment, and drug user status.ConclusionAs the first bioinformatic report on HIV-integrase from Iran, this study indicates that EVG and RAL are the optimal INTIs in first-line antiretroviral therapy (ART) in Iranian patients. Some conserved motifs and specific amino acids in INT-protein binding sites have characterized that mutation(s) in them may disrupt INT-drugs interaction and cause a significant loss in susceptibility to INTIs. Good adherence, treatment of naïve patients, and monitoring injection drug users are fundamental factors to control HIV infection in Iran effectively.     

A segmentation approach to long duration surface EMG recordings.

The purpose of this study was to develop an automatic segmentation method in order to identify postural surface EMG segments in long-duration recordings. Surface EMG signals were collected from the cervical erector spinae (CES), erector spinae (ES), external oblique (EO), and tibialis anterior (TA) muscles of 11 subjects using a bipolar electrode configuration. Subjects remained seated in a car seat over the 150-min data-collection period. The modified dynamic cumulative sum (MDCS) algorithm was used to automatically segment the surface EMG signals. Signals were rejected by comparison with an exponential mathematical model of the spectrum of a surface EMG signal. The average power ratio computed between two successive retained segments was used to classify segments as postural or surface EMG. The presence of a negative slope of a regression line fitted to the median frequency values of postural surface EMG segments was taken as an indication of fatigue. Alpha level was set at 0.05. The overall classification error rate was 8%, and could be performed in 25 min for a 150-min signal using a custom-built software program written in C (Borland Software Corporation, CA, USA). This error rate could be enhanced by concentrating on the rejection method, which caused most of the misclassification (6%). Furthermore, the elimination of non-postural surface EMG segments by the use of a segmentation approach enabled muscular fatigue to be identified in signals that contained no evidence of fatigue when analysed using traditional methods.     

Ovicidal activity of Sesamum indicum (L.) oil against Spodoptera littoralis (Boisd.)

In the research for alternative tools and botanical products to control Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae). Sesamum indicum (L.) (Lamiales: Pedaliaceae) oil was assayed as an ovicide. The mortality increased with existence of fatty acids. Chemical analysis of S. indicum oil using GLC analysis showed palmitic acid as the major fatty acid (51.27%), while the major hydrocarbon and sterols were found to be heneicosane (58.63%) and β-sitosterol (2.60%), respectively. Generally, the values of LC₅₀s indicated that one-day-old egg masses are more susceptible than three-day-old eggs. Also, the leaf dip technique is more efficient than the spraying one. Results showed several features of chorionic surface deformation treated with sesame and KZ oils than control using scanning electron microscopy. Meanwhile, the tested oils caused significant reduction in both total soluble protein and transaminase enzymes as compared to control. Additionally, the oils elongated the incubation period and larval duration than control.     

A Novel Molecular Design for a Hybrid Phage-DNA Construct Against DKK1

Nucleic acid immunization has recently exhibited a great promise for immunotherapy of various diseases. However, it is now clear that powerful strategies are imminently needed to improve their efficiency. In this regard, whole bacteriophage particles have been described as efficient DNA vaccine delivery vehicles, capable of circumventing the limitations of naked DNA immunization. Moreover, phage particles could be engineered to display specific peptides on their surfaces. Given these inherent characteristics of phages, we have designed a novel hybrid phage-DNA immunization vector using both M13 and pAAV plasmid elements. Following the construction and in vitro confirmation of the designed vectors, they were used for comparative mice immunization, carrying the same DNA sequence. The results indicated the efficacy of the designed hybrid phage particles, to elicit higher humoral immunity, in comparison to conventional DNA-immunization vectors (pCI). In light of these findings, it could be concluded that using adeno-associated virus (AAV) expression cassette along with displaying TAT peptide on the surface of the phage particle could be deemed as an appealing strategy to enhance the DNA-immunization and vaccination efficacy.     

Genetically encoded FRET-based optical sensor for Hg2+ detection and intracellular imaging in living cells

Journal of Industrial Microbiology & Biotechnology - Due to the potential toxicity of mercury, there is an immediate need to understand its uptake, transport and flux within living cells....     

The effect of redox signaling on extracellular matrix changes in diabetic wounds leading to amputation

Introduction& Objectives: Redox signaling is a critical regulator in the process of wound healing. This signaling pathway can be effective in the development or healing of diabetic ulcers through the ECM.In this study, the structure of extracellular matrix investigated in relation to redox signaling in the tissue of patients with diabetic ulcers that lead to organ amputation.Materials and methodsThe case-control design on diabetic patients ulcers as case group and non-diabetic limb ischemia as control were used.Hematoxylin-eosin, trichrome, and elastin staining methods were used for pathological evaluations of ECM. MDA, total thiol, and SOD levels were measured using ELISA kits to assess the oxidative stress level. Also, NO level was measured by using ELISA kits in both groups. Expression levels of genes MMP2, MMP9, and HIF were detected using real-time PCR with SYBR-green assay.ResultsThe pathological results showed an increase in the thickness of collagen and elastin fibers. Lipids atrophy was visible in the tissue isolated from the diabetic wound group. The amount of MAD to evaluate the level of lipid oxidation in patients with diabetic Ulcer was significantly higher than the control group(p < 0.01). Thiol level was significantly lower in the diabetic ulcer group than in the control group(p < 0.0001). The expression of metalloproteinases 2 and 9 genes in the tissues isolated from diabetic ulcers was lower than the control group(p < 0.0001). While the expression of the HIF gene in this group was higher than the control group(p < 0.0001).ConclutionIn the diabetic wound, the HIF secretion due to hypoxic conditions is beneficial for matrix deposition and prevents protease activity, but if the hypoxia persists, it can lead to ECM deposition subsequently increases the tissue pressure, increases of the collagen I-to-collagen III ratio in collagen accumulation that due to more hypoxia , lipidsAtrophy and eventually amputation.     

Expression of tomato thymidine kinase 1 by means of the baculovirus expression vector system

ABSTRACT

Tomato thymidine kinase 1 (ToTK1) is a deoxyribonucleoside kinase (dNK) that has been subject to study because of its potential to phosphorylate the nucleoside analogue 3-azido-2,3-dideoxythymidine (azidothymidine, AZT) equally well as its natural substrate thymidine (dThd). The combination of ToTK1 and AZT has been tested in two animal studies for its efficiency and use in suicide gene therapy for malignant glioma. The determination of the 3D structure of ToTK1 might shed light on the structure–function relationships of nucleoside activation by this enzyme and thereby show routes toward further improvement of ToTK1 and other TK1-like dNKs for suicide gene therapy. Here we report the successful expression of both full-length ToTK1 and a C-terminal truncated ToTK1 in Spodoptera frugiperda and Trichoplusia ni insect cells using the baculovirus expression vector system. This constitutes a further step on the road to determine the 3D structure of the first TK1 of plant origin, but also an enzyme with great potential for dNK-mediated suicide gene therapy.     

NSort/DB:An Intranuclear Compartment Protein Database

Distinct substructures within the nucleus are associated with a wide variety of important nuclear processes.Structures such as chromatin and nuclear pores have specific roles,while others such as Cajal bodies are more functionally varied.Understanding the roles of these membraneless intra-nuclear compartments requires extensive data sets covering nuclear and compartment-associated proteins.NSort/DB is a database providing access to intra-or sub-nuclear compartment associations for the mouse nuclear proteome.Based on resources ranging from large-scale curated data sets to detailed experiments,this data set provides a high-quality set of annotations of non-exclusive association of nuclear proteins with structures such as promyelocytic leukaemia bodies and chromatin.The database is searchable by protein identifier or compartment,and has a documented web service API.The search interface,web service and data download are all freely available online at http://www.nsort.org/db/.Availability of this data set will enable systematic analyses of the protein complements of nuclear compartments,improving our understanding of the diverse functional repertoire of these structures.