Controlled adhesion and growth of long term glial and neuronal cultures on Parylene-c

This paper explores the long term development of networks of glia and neurons on patterns of Parylene-C on a SiO 2 substrate. We harvested glia and neurons from the Sprague-Dawley (P1-P7) rat hippocampus and utilized an established cell patterning technique in order to investigate cellular migration, over the course of 3 weeks. This work demonstrates that uncontrolled glial mitosis gradually disrupts cellular patterns that are established early during culture. This effect is not attributed to a loss of protein from the Parylene-C surface, as nitrogen levels on the substrate remain stable over 3 weeks. The inclusion of the anti-mitotic cytarabine (Ara-C) in the culture medium moderates glial division and thus, adequately preserves initial glial and neuronal conformity to underlying patterns. Neuronal apoptosis, often associated with the use of Ara-C, is mitigated by the addition of brain derived neurotrophic factor (BDNF). We believe that with the right combination of glial inhibitors and neuronal promoters, the Parylene-C based cell patterning method can generate structured, active neural networks that can be sustained and investigated over extended periods of time. To our knowledge this is the first report on the concurrent application of Ara-C and BDNF on patterned cell cultures. © 2011 Delivopoulos, Murray.

RNAi, DRD1, and histone methylation actively target developmentally important Non-CG DNA methylation in Arabidopsis

Cytosine DNA methylation protects eukaryotic genomes by silencing transposons and harmful DNAs, but also regulates gene expression during normal development. Loss of CG methylation in the Arabidopsis thaliana met1 and ddm1 mutants causes varied and stochastic developmental defects that are often inherited independently of the original met1 or ddm1 mutation. Loss of non-CG methylation in plants with combined mutations in the DRM and CMT3 genes also causes a suite of developmental defects. We show here that the pleiotropic developmental defects of drm1 drm2 cmt3 triple mutant plants are fully recessive, and unlike phenotypes caused by met1 and ddm1, are not inherited independently of the drm and cmt3 mutations. Developmental phenotypes are also reversed when drm1 drm2 cmt3 plants are transformed with DRM2 or CMT3, implying that non-CG DNA methylation is efficiently re-established by sequence-specific signals. We provide evidence that these signals include RNA silencing though the 24-nucleotide short interfering RNA (siRNA) pathway as well as histone H3K9 methylation, both of which converge on the putative chromatin-remodeling protein DRD1. These signals act in at least three partially intersecting pathways that control the locus-specific patterning of non-CG methylation by the DRM2 and CMT3 methyltransferases. Our results suggest that non-CG DNA methylation that is inherited via a network of persistent targeting signals has been co-opted to regulate developmentally important genes. © 2006 Chan et al.

A preliminary study on the protein requirement of Chanos chanos (Forskal) fry in a controlled environment

Growth rate of fish appeared to be related to the levels of the protein in the diet up to 40%. Fish fed diets containing 50 and 60% grew slower than those fed 40%, and the optimum level appears to be 40% when fed to fry at a rate of 10% of body weight. Best feed conversion of 1.96 was also obtained from the 40% protein diet. Mean survival rates were low in all treatments, but highest for the 40% protein diet. The competition of 5 isocaloric experimental diets containing various levels of protein are tabulated, as are weight gains, diet conversions and survival rates for milkfish fry fed various dietary levels of protein. Growth curves for milkfish fry are shown, and the relationship between weight gains of milkfish fry and the dietary levels of protein are illustrated.

How to avoid the problems of target-setting

Purpose: Advocates and critics of target-setting in the workplace seem unable to reach beyond their own well-entrenched battle lines. While the advocates of goal-directed behaviour point to what they see as demonstrable advantages, the critics of target-setting highlight equally demonstrable disadvantages. Indeed, the academic literature on this topic is currently mired in controversy, with neither side seemingly capable of envisaging a better way forward. This paper seeks to break the current deadlock and move thinking forward in this important aspect of performance measurement and management by outlining a new, more fruitful approach, based on both theory and practical experience. Design/methodology/approach: The topic was approached in three phases: assembling and reading key academic and other literature on the subject of target-setting and goal-directed behaviour, with a view to understanding, in depth, the arguments advanced by the advocates and critics of target-setting; comparing these published arguments with one's own experiential findings, in order to bring the essence of disagreement into much sharper focus; and then bringing to bear the academic and practical experience to identify the essential elements of a new, more fruitful approach offering all the benefits of goal-directed behaviour with none of the typical disadvantages of target-setting. Findings: The research led to three key findings: the advocates of goal-directed behaviour and critics of target-setting each make valid points, as seen from their own current perspectives; the likelihood of these two communities, left to themselves, ever reaching a new synthesis, seems vanishingly small (with leading thinkers in the goal-directed behaviour community already acknowledging this); and, between the three authors, it was discovered that their unusual combination of academic study and practical experience enabled them to see things differently. Hence, they would like to share their new thinking more widely. Research limitations/implications: The authors fully accept that their paper is informed by extensive practical experience and, as yet, there have been no opportunities to test their findings, conclusions and recommendations through rigorous academic research. However, they hope that the paper will move thinking forward in this arena, thereby informing future academic research. Practical implications: The authors hope that the practical implications of the paper will be significant, as it outlines a novel way for organisations to capture the benefits of goal-directed behaviour with none of the disadvantages typically associated with target-setting. Social implications: Given that increased efficiency and effectiveness in the management of organisations would be good for society, the authors think the paper has interesting social implications. Originality/value: Leading thinkers in the field of goal-directed behaviour, such as Locke and Latham, and leading critics of target-setting, such as Ordóñez et al. continue to argue with one another - much like, at the turn of the nineteenth century, proponents of the "wave theory of light" and proponents of the "particle theory of light" were similarly at loggerheads. Just as this furious scientific debate was ultimately resolved by Taylor's experiment, showing that light could behave both as a particle and wave at the same time, the authors believe that the paper demonstrates that goal-directed behaviour and target-setting can successfully co-exist. © Emerald Group Publishing Limited.

Calcium-permeable acid-sensing ion channel is a molecular target of the neurotoxic metal ion lead

Acid-sensing ion channels (ASICs) are emerging as fundamental players in the regulation of neural plasticity and in pathological conditions. Here we showed that lead (Pb2+), a well known neurotoxic metal ion, reversibly and concentration-dependently inhib

The genetic diversity of Acipenser stellatus populations in the Caspian Sea was studied using microsatellite technique

A total of 361 caudal fin samples were collected from adult A. stellatus specimens caught in the north Caspian Sea, including specimens from Kazakhstan (Ural River), Russia (Volga River), Azerbaijan (Kura River), specimens caught in the south Caspian Sea including specimens from Fishery Zone 1 (from Astara to Anzali), Fishery Zone 2 (from Anzali to Ramsar), Fishery Zone 3 (from Nowshahr to Babolsar), Fishery Zone 4 (from Miyankaleh to Gomishan) as well as from specimens caught in Turkmenistan (all specimens were collected during the sturgeon stock assessment survey). About 2 g of fin tissue was removed from each caudal fin sample, stored in 96% ethyl alcohol and transferred to the genetic laboratory of the International Sturgeon Research Institute. Genomic DNA was extracted using phenol-chloroform method. The quality and quantity of DNA was assessed using 1% Agarose gel electrophoresis and Polymerase Chain Reaction (PCR) was conducted on the target DNA using 15 paired microsatellite primer. PCR products were electrophoresed on polyacrylamide gels (6%) that were stained using silver nitrate. Electrophoretic patterns and DNA bands were analyzed with BioCapt software. Allele count and frequency, genetic diversity, expected heterozygosity and observed heterozygosity allele number, and the effective allele number, genetic similarity and genetic distance, FST and RST were calculated. The Hardy Wienberg Equilibrium based on X2 and Analysis of Molecular Variance (AMOVA) at 10% confidence level was calculated using the Gene Alex software. Dendrogram for genetic distances and identities were calculated using TFPGA program for any level of the hierarchy. It is evident from the results obtained that the 15 paired primers studied, polymorphism was observed in 10 pairs in 12 loci, while one locus did not produce DNA bands. Mean allele number was 13.6. Mean observed and expected heterozygosity was 0.86 and 0.642, respectively. It was also seen that specimens from all regions were not in Hardy Wienberg Equilibrium in most of the loci (P≤0.001). Highest Fst (0.063) was observed when comparing specimens from Fishery Zone 2 and Fishery Zone 4 (Nm=3.7) and lowest FST (0.028) was observed when comparing specimens from the Volga River and those from the Ural River (8.7). Significant differences (P<0.01) were observed between RST recorded in the specimens studied. Highest genetic distance (0.604) and lowest genetic resemblance (0.547) were observed between specimens from Fishery zones 2 and 4. Lowest genetic distance (0.311) and highest genetic resemblance (0.733) was observed between specimens from Turkmenistan and specimens from Fishery zone 1. Based on the genetic dendrogeram tree derived by applying UPGMA algorithm, A. stellatus specimens from Fishery zone 2 or in other words specimens from the Sepidrud River belong to one cluster which divides into two clusters, one of which includes specimens from Fishery zones 1, 3 and 4 and specimens from Turkmenistan while the other cluster includes specimens from Ural, Volga and Kura Rivers. It is thus evident that the main population of this species belongs to the Sepidrud River. Results obtained from the present study show that at least eight different populations of A. stellatus are found in the north and south Caspian Sea, four of which are known populations including the Ural River population, the Volga River population, the Kura River population and the Sepidrud River populations. The four other populations identified belonging to Fishery zones 1, 3, and 4 and to Turkmenistan are most probably late or early spawners of the spring run and autumn run of each of the major rivers mentioned. Specific markers were also identified for each of the populations identified. The Ural River population can be identified using primers Spl-68, 54b and Spl-104, 163 170, 173, the Volga River population can be identified using primers LS-54b and Spl-104, 170, 173 113a and similarly the population from the Kura River can be identified using primers LS-34, 54b and Spl-163, 173 and that from the Sepidrud River can be identified using primers LS-19, 34, 54b and Spl-105, 113b. This study gives evidence of the presence of different populations of this species and calls for serious measures to be taken to protect the genetic stocks of these populations. Considering that the population of A. stellatus in Fishery zone 2 is an independent population of the Sepidrud River in the Gilan Province, the catch of these fishes in the region needs to be controlled and regulated in order to restore the declining stocks of this species.

Identification of triosephosphate isomerase (TIM) genes from Microcystis (Cyanobacteria : Chroococcales) and theoretical analyses of the potential of cyanobacterial TIM as a target for designing specific inhibitors

The genes encoding triosephosphate isomerase (TIM) in three species of Microcystis (M. aeruginosa, M. viridis and M. wesenbergii) were investigated. Reverse transcriptase-polymerase chain reaction indicated that they were transcribed in the cells. Analyses showed that their DNA and deduced amino acid sequences were highly conserved between all the three species, only a single nonsynonymous substitution was seen at position 31, from an Asp in M. aeruginosa and M. viridis to Glu in M. wesenbergii. Sequence alignment of these with 12 other known cyanobacterial TIM sequences showed that all the cyanobacterial TIMs had a very high level of amino acid identity (over 50% between each two). Comparison of the cyanobacterial TIMs with other reported TIMs (from diverse lineages of the three Domains) showed that they possessed common active-site residues and sequence motifs. All cyanobacterial TIMs have two common cysteine residues (Cys127 and Cys176), and the Cys176 is almost cyanobacteria-specific with only one exception in Streptomyces coelicolor. Both secondary structure alignment and comparative modelling of Synechocystis sp. TIM showed that Cys176 was located at the hinge region of the flexible loop-6 and might therefore be critical to the movement of TIM's loop-6, which is important to the function of the enzyme. Thus, the cyanobacterial TIM-specific Cys176 may be a potential site for the discovery of suitable drugs against cyanobacteria, and such drugs may have utility in controlling water blooms due to cyanobacteria.

Developing a technology intelligence strategy at kodak european research: Scan & target

OVERVIEW: Kodak European Research (KER) developed a strategy for technology intelligence based on a theoretical model developed by Kerr et al. (2006). KER scouts designed and implemented a four-step approach to identify relevant technologies and research centers across Europe, Africa and the Middle East. The approach provides clear guidance for integrating web searches, scouting trips, networking and interactions with intermediaries. KER's example illustrates how companies can organize themselves to look outside corporate boundaries in search of technologies relevant for their business. The approach may be useful to those in other companies who have been asked to start a technology intelligence activity. © 2010 Industrial Research Institute, Inc.

The morphology of decorated amyloid fibers is controlled by the conformation and position of the displayed protein.

Self-assembled structures capable of mediating electron transfer are an attractive scientific and technological goal. Therefore, systematic variants of SH3-Cytochrome b(562) fusion proteins were designed to make amyloid fibers displaying heme-b(562) electron transfer complexes. TEM and AFM data show that fiber morphology responds systematically to placement of b(562) within the fusion proteins. UV-vis spectroscopy shows that, for the fusion proteins under test, only half the fiber-borne b(562) binds heme with high affinity. Cofactor binding also improves the AFM imaging properties and changes the fiber morphology through changes in cytochrome conformation. Systematic observations and measurements of fiber geometry suggest that longitudinal registry of subfilaments within the fiber, mediated by the interaction and conformation of the displayed proteins and their interaction with surfaces, gives rise to the observed morphologies, including defects and kinks. Of most interest is the role of small molecule modulation of fiber structure and mechanical stability. A minimum complexity model is proposed to capture and explain the fiber morphology in the light of these results. Understanding the complex interplay between these factors will enable a fiber design that supports longitudinal electron transfer.

The application of STEM and in situ controlled dehydration to bacterial systems using ESEM.

Transmission imaging with an environmental scanning electron microscope (ESEM) (Wet STEM) is a recent development in the field of electron microscopy, combining the simple preparation inherent to ESEM work with an alternate form of contrast available through a STEM detector. Because the technique is relatively new, there is little information available on how best to apply this technique and which samples it is best suited for. This work is a description of the sample preparation and microscopy employed by the authors for imaging bacteria with Wet STEM (scanning transmission electron microscopy). Three different bacterial samples will be presented in this study: first, used as a model system, is Escherichia coli for which the contrast mechanisms of STEM are demonstrated along with the visual effects of a dehydration-induced collapse. This collapse, although clearly in some sense artifactual, is thought to lead to structurally meaningful morphological information. Second, Wet STEM is applied to two distinct bacterial systems to demonstrate the novel types of information accessible by this approach: the plastic-producing Cupriavidus necator along with wild-type and ΔmreC knockout mutants of Salmonella enterica serovar Typhimurium. Cupriavidus necator is shown to exhibit clear internal differences between bacteria with and without plastic granules, while the ΔmreC mutant of S. Typhimurium has an internal morphology distinct from that of the wild type.

Improving model quality through computer controlled saturation

In dynamic centrifuge modelling, fluids with enhanced viscosity are often used to correct for the discrepancy in time scaling. However, great care must be taken when using a high viscosity fluid that hydraulic gradients during saturation do not become high enough to cause excessive model disturbance. This paper introduces the CAM-Sat system which aims to improve the saturation process by continually controlling the fluid flow into the model, limiting it to rates low enough to avoid model disturbance. A new method for measuring the fluid flow rate is then described, and its implementation & improvement to the system is discussed. © 2010 Taylor & Francis Group, London.