Foliar application of ascorbic and citric acids enhanced ‘Red Spur’ apple fruit quality,bioactive compounds and antioxidant activity

This study aimed to assess the effect of the foliar application of ascorbic acid (AA) and citric acid (CA) on total antioxidant activity (TAA), total phenolics, total flavonoids, total anthocyanin content, antioxidant enzymes, phenylalanine ammonialyase (PAL), and polyphenol oxidase (PPO) activities in apple ‘Red Spur’. The experiment was conducted on 12-years-old trees ‘Red Spur’ grafted on MM₁₀₆ rootstock. The trees were sprayed with AA (0, 200 and 400 mg L^?1) and/or CA (0, 200 and 400 mg L^?1) at three different times during summer. Foliar application with AA and CA significantly (p < 0.01) enhanced all measured quality attributes and decreased the activity of PPO. Fruit from trees treated with AA at 400 mg L^?1 and CA at 200 mg L^?1 showed the highest TAA and catalase (CAT) enzyme activity. Total phenolics increased in fruits when trees were sprayed with AA and CA. Contrasting, AA treatment, CA had no significant effect on guaiacol peroxidase (G-POD). A significant decrease in PPO activity was detected in fruits when treated with both AA and CA. Both treatments significantly decreased the activity of PAL at 400 mg L^?1. Considering the results, foliar application of AA and CA, either alone or in combination improved the quality and nutraceutical properties of ‘Red Spur’ apple.     

<Emphasis Type="Italic">BMP3</Emphasis> promoter hypermethylation in plasma-derived cell-free DNA in colorectal cancer patients

Detecting cfDNA in plasma or serum could serve as a ‘liquid biopsy’, for circulating tumor DNA with aberrant methylation patterns offer a possible method for early detection of several cancers which could avoid the need for tumor tissue biopsies. Bone Morphogenetic Protein 3 (BMP3) was identified as a candidate tumor suppressor gene putatively down-regulated in colorectal cancer (CRC). In this study, we aimed to assess the potential role of BMP3 promoter methylation changes in plasma DNA for detection of colorectal cancerous and precancerous lesions. Plasma DNA samples were extracted from 50 patients with histologically diagnosed polyps or tumor and 50 patients reported negative for polyps or tumors. The procedure consists of bisulfite conversion of the extracted DNA, purification of bis-DNA, and BMP3 methylation status analysis by using the bisulfite specific high resolution melting analysis. This study demonstrated that there was a significantly higher frequency of BMP3 methylated DNA in plasma in patients with polyps versus healthy controls with a sensitivity and specificity of 40 and 94%, respectively. In conclusion, our results demonstrated that BMP3 DNA methylation in plasma had not have sufficient sensitivity and it should be used in combination with other biomarkers for the detection of CRC.     

Expression of <Emphasis Type="Italic">GNA</Emphasis> and biting site-restricted <Emphasis Type="Italic">cry1Ac</Emphasis> in cotton; an efficient attribution to insect pest management strategies

Insect-resistant transgenic cotton has been commercialized for two decades. Most of the introduced cultivars express Bt gene(s) constitutively under the control of 35S promoter in whole-plant tissues. However, there have been other promoters considered by researchers to confine the toxin expression to targeted organ and tissues. We developed a triple-gene construct including GNA, cry1Ac and cp4 epsps genes. We attempted to confine cry1Ac expression to insect biting sites by cloning it to downstream of a wound-inducible promoter isolated from Asparagus officinalis (AoPR1). Moreover, to broaden the range of resistance, GNA was driven by the 35S promoter to target the sap-sucking insects like aphids which impose large losses in cotton production. To select the transformants in selection medium and for glyphosate tolerance, GNA and cry1Ac genes were accompanied with cp4 epsps gene. Two binary vectors harboring desired genes were constructed and utilized in the study (pGTGNAoC1AC and pGTGN35C1AC). Transformation of cultivar GSN-12 was carried out by employing Agrobacterium tumefaciens strain EHA105. Plantlets were primarily screened under glyphosate (N-phosphonomethyl glycine) selection pressure and subsequently subjected to molecular and biotoxicity assays. Introduction of cry1Ac and GNA to cotton plant conferred resistance to Spodoptera littoralis and Aphis gossypii Glover. Restriction of cry1Ac toxin protein to insect biting sites along with a plant lectin attributes significantly to insect pest management strategies.     

Dismissal of Acoelomorpha: Acoela and Nemertodermatida are separate early bilaterian clades

We used new 18S and 28S rRNA sequences analysed with parsimony, maximum likelihood and Bayesian methods of phylogenetic reconstruction to show that Nemertodermatida, generally classified as the sister group of Acoela within the recently proposed Phylum Acoelomorpha, are a separate basal bilaterian lineage. We used several analytical approaches to control for possible long branch attraction (LBA) artefacts in our results. Parsimony and the model based phylogenetic reconstruction methods that incorporate 'corrections' for substitution rate heterogenities yielded concordant results. When putative long branch taxa were experimentally removed the resulting topologies were consistent with our total evidence analysis. Deletion of fast-evolving nucleotide sites decreased resolution and clade support, but did not support a topology conflicting with the total evidence analysis. Establishment of Acoela and Nemertodermatida as two early lineages facilitates reconstruction of ancestral bilaterian features. The ancestor of extant Bilateria was a small, benthic direct developer without coelom or a planktonic larval stage. The previously proposed Phylum Acoelomorpha is dismissed as paraphyletic.     

Simultaneous electrochemical determination of l-cysteine and l-cysteine disulfide at carbon ionic liquid electrode

A linear sweep voltammetric method is used for direct simultaneous determination of l-cysteine and l-cysteine disulfide (cystine) based on carbon ionic liquid electrode. With carbon ionic liquid electrode as a high performance electrode, two oxidation peaks for l-cysteine (0.62 V) and l-cysteine disulfide (1.3 V) were observed with a significant separation of about 680 mV (vs. Ag/AgCl) in phosphate buffer solution (pH 6.0). The linear ranges were obtained as 1.0–450 and 5.0–700 μM and detection limits were estimated to be 0.298 and 4.258 μM for l-cysteine and l-cysteine disulfide, respectively. This composite electrode was applied for simultaneous determination of l-cysteine and l-cysteine disulfide in two real samples, artificial urine and nutrient broth. Satisfactory results were obtained which clearly indicate the applicability of the proposed electrode for simultaneous determination of these compounds in complex matrices.     

The Degree of Resistance of Erythrocyte Membrane Cytoskeletal Proteins to Supra-Physiologic Concentrations of Calcium: An In Vitro Study

Calcium is a key regulator of cell dynamics. Dysregulation of its cytosolic concentration is implicated in the pathophysiology of several diseases. This study aimed to assess the effects of calcium on the network of membrane cytoskeletal proteins. Erythrocyte membranes were obtained from eight healthy donors and incubated with 250 µM and 1.25 mM calcium solutions. Membrane cytoskeletal proteins were quantified using SDS-PAGE at baseline and after 3 and 5 days of incubation. Supra-physiologic concentrations of calcium (1.25 mM) induced a significant proteolysis in membrane cytoskeletal proteins, compared with magnesium (p < 0.001). Actin exhibited the highest sensitivity to calcium-induced proteolysis (6.8 ± 0.3 vs. 5.3 ± 0.6, p < 0.001), while spectrin (39.9 ± 1.0 vs. 40.3 ± 2.0, p = 0.393) and band-6 (6.3 ± 0.3 vs. 6.8 ± 0.8, p = 0.191) were more resistant to proteolysis after incubation with calcium in the range of endoplasmic reticulum concentrations (250 µM). Aggregation of membrane cytoskeletal proteins was determined after centrifugation and was significantly higher after incubation with calcium ions compared with control, EDTA and magnesium solutions (p < 0.001). In a supra-physiologic range of 1.25–10 mM of calcium ions, there was a nearly perfect linear relationship between calcium concentration and aggregation of erythrocyte membrane cytoskeletal proteins (R ² = 0.971, p < 0.001). Our observation suggests a strong interaction between calcium ions and membrane cytoskeletal network. Cumulative effects of disrupted calcium homeostasis on cytoskeletal proteins need to be further investigated at extended periods of time in disease states.     

γ-turn types prediction in proteins using the two-stage hybrid neural discriminant model

Due to the slightly success of protein secondary structure prediction using the various algorithmic and non-algorithmic techniques, similar techniques have been developed for predicting γ-turns in proteins by Kaur and Raghava [2003. A neural-network based method for prediction of γ-turns in proteins from multiple sequence alignment. Protein Sci. 12, 923-929]. However, the major limitation of previous methods was inability in predicting γ-turn types. In a recent investigation we introduced a sequence based predictor model for predicting γ-turn types in proteins [Jahandideh, S., Sabet Sarvestani, A., Abdolmaleki, P., Jahandideh, M., Barfeie, M, 2007a. γ-turn types prediction in proteins using the support vector machines. J. Theor. Biol. 249, 785-790]. In the present work, in order to analyze the effect of sequence and structure in the formation of γ-turn types and predicting γ-turn types in proteins, we applied novel hybrid neural discriminant modeling procedure. As the result, this study clarified the efficiency of using the statistical model preprocessors in determining the effective parameters. Moreover, the optimal structure of neural network can be simplified by a preprocessor in the first stage of hybrid approach, thereby reducing the needed time for neural network training procedure in the second stage and the probability of overfitting occurrence decreased and a high precision and reliability obtained in this way.     

RGD conjugation to polyethyleneimine does not improve DNA delivery to bone marrow stromal cells

Bone marrow stromal cells (BMSC) modified with therapeutic genes are being actively pursued for gene therapy protocols. To develop safe and effective nonviral methods for BMSC modification, the cationic polymer polyethyleneimine (PEI) has been utilized to condense plasmid DNA for intracellular delivery. This study was conducted to explore the feasibility of increasing the PEI's effectiveness by coupling integrin-binding arginine-glycine-aspartic acid (RGD) peptides to the polymer. BMSC from rats were isolated and expanded in culture for gene transfer studies. In contrast to our expectations, RGD-conjugated PEI did not exhibit an enhanced binding to BMSC. This was the case where the peptides were conjugated to PEI by short, disulfide linkages or long poly(ethylene glycol) linkages. Using a reporter gene for the enhanced green fluorescent protein, the transfection efficiency of RGD-conjugated PEI was also lower than the delivery by the native PEI, which exhibited equivalent transfection efficiency to that of an adenovirus. We conclude that native PEI was sufficient for the transformation of BMSC and that coupling of the integrin-binding RGD-peptides did not improve the effectiveness of this polymer for BMSC transfection.     

Transient Global Amnesia after Ablation of Premature Ventricular Beats Arising from the Right Coronary Cusp

A 58-year-old female underwent PVC ablation within the right coronary cusp for symptomatic PVCs and suspected PVC-induced cardiomyopathy. Immediately after the procedure, she started to complain about feelings of impending doom, disorientation to time and place, and amnesia regarding the procedure. No sensory or motor deficits could be elicited. A thromboembolic event was suspected and she was evaluated by a neurologist. CT scan of her brain was negative. She was diagnosed with transient global amnesia and her mentation returned to baseline within 4 hours after the procedure.     

Cancer and diet: How are they related?

Extensive research in the past decade has revealed cancer to be a multigenic disease caused by perturbation of multiple cell signalling pathways and dysregulation of numerous gene products, all of which have been linked to inflammation. It is also becoming evident that various lifestyle factors, such as tobacco and alcohol use, diet, environmental pollution, radiation and infections, can cause chronic inflammation and lead to tumourigenesis. Chronic diseases caused by ongoing inflammation therefore require chronic, not acute, treatment. Nutraceuticals, compounds derived from fruits, vegetables, spices and cereals, can be used chronically. This study discusses the molecular targets of some nutraceuticals that happen to be markers of chronic inflammation and how they can prevent or treat cancer. These naturally-occurring agents in the diet have great potential as anti-cancer drugs, thus proving Hippocrates, who proclaimed 25 centuries ago, 'Let food be thy medicine and medicine be thy food'.