Exploring the interaction between small RNAs and R genes during Brachypodium response to Fusarium culmorum infection

The present study aims to investigate small RNA interactions with putative disease response genes in the model grass species Brachypodium distachyon. The fungal pathogen Fusarium culmorum (Fusarium herein) and phytohormone salicylic acid treatment were used to induce the disease response in Brachypodium. Initially, 121 different putative disease response genes were identified using bioinformatic and homology based approaches. Computational prediction was used to identify 33 candidate new miRNA coding sequences, of which 9 were verified by analysis of small RNA sequence libraries. Putative Brachypodium miRNA target sites were identified in the disease response genes, and a subset of which were screened for expression and possible miRNA interactions in 5 different Brachypodium lines infected with Fusarium. An NBS-LRR family gene, 1g34430, was polymorphic among the lines, forming two major genotypes, one of which has its miRNA target sites deleted, resulting in altered gene expression during infection. There were siRNAs putatively involved in regulation of this gene, indicating a role of small RNAs in the B. distachyon disease response.     

Fabrication of a highly sensitive disposable immunosensor based on indium tin oxide substrates for cancer biomarker detection

Anti-HER-3 antibody was used for the first time in a disposable immunosensor based on indium tin oxide (ITO) substrate for HER-3 quantification. Anti-HER-3 was immobilized onto ITO substrate by 3-aminopropyl)triethoxysilane (APTES) and glutaraldehyde. This highly sensitive immunosensor was capable of detecting concentrations of HER-3 down to the femtogram/ml level by investigating changes in the charge transfer resistance (R_ct) using electrochemical impedance spectroscopy (EIS). Construction of ITO layers was carefully investigated using a broad range of techniques such as voltammetry, EIS, atomic force microscopy (AFM), and scanning electron microscopy (SEM). Meanwhile, in an immunosensor system, the “single frequency impedance” technique was first used for characterization of interaction between HER-3 and anti-HER-3. Eventually, the proposed ITO-based immunosensor was applied to artificial serum samples spiked with HER-3.     

Application of Partially Balanced Block Designs to Factorial Experiments

In this paper we present partially balanced block designs with F_p and C_p association schemes. These designs play a great role in the experimental theory since they can be applied to factorial experiments. A particular stress is put on the formulation of association schemes _p and C_p, presentation of exemplary constructions of these designs and their analysis of variance. The presented considerations are abundantly illustrated with examples.     

Analyses of the stomach deposit that develops in Myzus persicae feeding on sugar beet

Abstract .The white deposit, commonly found in the stomach of aphids that feed on sugar beet ( Beta vulgaris ssp. saccharifera L), turns a dark colour just prior to the death of the aphid, suggesting that chemical changes causing the deposit to darken may be associated with the death of the aphid. Chemical analyses of the white and dark stomach deposits by HPLC after hydrolysis, elemental analysis and FAB-MS, showed that the dark deposit is formed after the loss of hydrogen and oxygen from the organic white deposit, and that it appears to be a complex that is not dominated by any given compound. These results contrast with those of previous studies, in which the white precipitate was described as a polysaccharide or a mucopolysaccharide.     

A simple shoot multiplication procedure using internode explants,and its application for particle bombardment and <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation in watercress

A shoot multiplication system derived from internode explants was investigated with the aim of improving genetic characteristics of watercress (Nasturtium officinale R. Br.). Internodes of ca. 1 cm excised from in vitro stock shoot culture were placed on half-strength Murashige and Skoog (MS) medium supplemented with 3 μM 2,4-dichlorophenoxyacetic acid as a pre-treatment. Laser scanning microscopy indicated clearly that the first sign of meristematic cell division could be seen after 1–2 days of pre-culture, and meristematic tissues multiplied along the vascular cambium of the internode segment during 7 days of culture. Multiple shoots could be obtained from more than 90% of the pre-treated explants when they were subsequently transferred to MS medium supplemented with 1 μM thidiazuron for 3 weeks. These findings indicate that pre-treatment of the internodes for 7 days promoted their capacity for organogenesis. Using this pre-treatment, frequent generation of transgenic watercress plants was achieved by adapting particle bombardment and Agrobacterium-mediated transformation techniques with a construct expressing a synthetic green florescent protein gene.     

Validation of image analysis for enzyme histochemical and immunocytochemical staining

Immunocytochemical and enzyme histochemical analyses of cells and tissues are used to detect changes in the extent of injury and the expression of various molecules. Image analysis quantitation offers an easier, more efficient technique to evaluate these changes. We studied the application of image analysis for evaluating enzyme histochemistry and immunocytochemistry of cells and tissues as a way to assess stroke. Using brain sections, we compared investigator and computer-generated image analysis of 2,3,5-triphenyltetrazolium chloride stained cerebral infarcts in rats subjected to 2 h middle cerebral artery occlusion and 22 h re-perfusion. Both methods documented the infarct volumes with a comparison of means of less then 5%. This suggests no difference between computer- and hand-calculated values. Computer-generated analysis was easier and faster to use. Using endothelial cell monolayers, immunocytochemical staining of a time course of heat shock protein expression was compared to a grading system using fast red chromagen counterstained with hematoxylin. Results demonstrated greater ease and efficiency with computer-generated image analysis compared to other subjective systems of analysis. Image analysis is more useful for detecting small differences in staining, especially when using 3,3-diaminobenzidine as a chromagen. Investigator bias is also reduced using this system. Our comparisons validate the use of this versatile technology to assess more easily both cell and tissues in stroke research.     

Extension of the Labeled Precursor Method of Measuring Neurotransmitter Kinetics to Non-Steady-State Conditions: Application to Striatal Dopamine Metabolism

Abstract: A solution of the differential equations describing the single open compartment model of neurotransmitter kinetics which does not assume a steady state is presented. In addition, a computer procedure is described for analyzing data obtained following the pulse injection of radiolabeled precursors, which eliminates the use of graphical methods in calculating fractional rate constants of synthesis and catabolism. This procedure also provides statistically valid estimates of the standard errors of the rate constants, allowing for tests of the significance of experimental effects. The ability of this approach to discriminate between changes in synthesis and catabolism under non-steadystate conditions is demonstrated by analyzing the kinetics of striatal dopamine metabolism following (a) administration of α-methyl-ρ-tyrosine and (b) lesion of the substantia nigra. The utility of this method and precautions in its use are discussed.     

Sampling error as a misleading artifact in “key factor analysis”

The influences of sampling error in key factor analysis are investigated statistically. The error involved in the data distorts the results in various misleading ways. In the course of detecting key factors by correlation analysis, the distortion arises in the following two ways: (1) the contributions made by the first and the last components of population trend index (log I or K) to the total variation are overrated as compared with the others; and (2) spurious negative correlation arises between successive two components. The risk of misinterpretation due to such disturbance is usually increased further if the error is concentrated on any particular developmental stages. In the tests to detect density-dependence by using regression analysis, the error consistently acts as if it were a density-dependent factor: under the effect of sampling error, the slope b for the regression of log N_i+1 on log N_i, for example, is expected to become<1 even where there is no density-dependent factor at all. A set of formulas are derived which may serve to check and correct these misleading distortions caused by the error. It is also shown that such undesirable influences can be avoided, at least to a considerable extent, if appropriate sampling plans are adopted for the study. The validity of key factor analysis is discussed in reference to this and some related problems.     

A New Statistical Method for Mixed Variates Analysis in Epidemiological Research

Epidemiologically oriented research often may not do without observational or only partially controlled studies. In many such situations both qualitative characteristics and quantitative ones are observed. In literature there are different methods of handling such problems. The paper presents a method for analyzing dependencies resp. associations between random variates of any kind. The model concerned fullfills the whole field between analysis of variance, analysis of covariance and contingency table analysis. The method is named MIVA or mixed variates analysis, bases on the class of Conditional Gaussian Distributions of the exponential family and results in a unique system of mixed and unmixed measures of association–of pairwise, partial, multiple and global type. These measures are easy to be estimated and tested on significant deviation from zero. They may be used describing or analyzing dependence structures in many epidemiological studies but also in other fields.