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    ABSTRACT: Sampling of various terrestrial habitats (woodland, grassland, sea edge) of Hawthorn Dene and surroundings within the Magnesian Limestone plateau of County Durham yielded 17 species of Enchytraeidae and the naidid/tubificid Rhyacodrilus falciformis. Five enchytraeid species were recorded for the first time in the British Isles: Fridericia auritoides, F. maculatiformis, F. semisetosa, F. sylvatica, and F. isseli.
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    ABSTRACT: Hydrodynamics, cell wall and ion dynamics are all important properties that regulate pollen tube growth. Currently, the two main pollen tube growth models, the cell wall model and the hydrodynamic model do not appear to be reconcilable. Here we develop an integrative model for pollen tube growth and show that our model reproduces key experimental observations: (1) that the hypertonic condition leads to a much longer oscillatory period and that the hypotonic condition halves the oscillatory period; (2) that oscillations in turgor are experimentally undetectable; (3) that increasing the extracellular calcium concentration or decreasing the pH decreases the growth oscillatory amplitude; (4) that knockout of Raba4d, a member of the Rab family of small GTPase proteins, decreases pollen tube length after germination for 24 h. Using the model generated here, we reveal that (1) when cell wall extensibility is large, pollen tube may sustain growth at different volume changes and maintain relatively stable turgor; (2) turgor increases if cell wall extensibility decreases; (3) increasing turgor due to decrease in osmolarity in the media, although very small, increases volume change. However, increasing turgor due to decrease in cell wall extensibility decreases volume change. In this way regulation of pollen tube growth by turgor is context dependent. By changing the osmolarity in the media, the main regulatory points are extracellular osmolarity for water flow and turgor for the volume encompassed by the cell wall. However, if the viscosity of cell wall changes, the main regulatory points are turgor for water flow and wall extensibility for the volume encompassed by the cell wall. The novel methodology developed here reveals the underlying context-dependent regulatory principle of pollen tube growth.
    Frontiers in Plant Science 08/2014; 5:392. DOI:10.3389/fpls.2014.00392
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    ABSTRACT: Ubiquitin is a peptide modifier able to form polymers of varying length and linkage as part of a powerful signaling system. Perhaps the best-known aspect of this protein's function is as the driver of targeted protein degradation through the Ubiquitin Proteasome System (UPS). Through the formation of lysine 48-linked polyubiquitin chains, it is able to direct the degradation of tagged proteins by the 26S proteasome, indirectly controlling many processes within the cell. However, recent research has indicated that ubiquitin performs a multitude of other roles within the cell beyond protein degradation. It is able to form 6 other "atypical" linkages though lysine residues at positions 6, 11, 27, 29, 33, and 63. These atypical chains perform a range of diverse functions, including the regulation of iron uptake in response to perceived deficiency, repair of double stranded breaks in the DNA, and regulation of the auxin response through the non-proteasomal degradation of auxin efflux carrier protein PIN1. This review explores the role ubiquitin chain topology plays in plant cellular function. We aim to highlight the importance of these varying functions and the future challenges to be encountered within this field.
    Frontiers in Plant Science 04/2014; 5:122. DOI:10.3389/fpls.2014.00122
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    ABSTRACT: Plants are sessile organisms and therefore they must adapt their growth and architecture to a changing environment. Understanding how hormones and genes interact to coordinate plant growth in a changing environment is a major challenge in developmental biology. Although a localized auxin concentration maximum in the root tip is important for root development, auxin concentration cannot change independently of multiple interacting hormones and genes. In this review, we discuss the experimental evidence showing that the POLARIS peptide of Arabidopsis plays an important role in hormonal crosstalk and root growth, and review the crosstalk between auxin and other hormones for root growth with and without osmotic stress. Moreover, we discuss that experimental evidence showing that, in root development, hormones and the associated regulatory and target genes form a network, in which relevant genes regulate hormone activities and hormones regulate gene expression. We further discuss how it is increasingly evident that mathematical modeling is a valuable tool for studying hormonal crosstalk. Therefore, a combined experimental and modeling study on hormonal crosstalk is important for elucidating the complexity of root development.
    Frontiers in Plant Science 03/2014; 5:116. DOI:10.3389/fpls.2014.00116
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    ABSTRACT: The atrial volume receptor reflex arc serves to regulate plasma volume. Atrial volume receptors located in the endocardium of the atrial wall undergo mechanical deformation as blood is returned to the atria of the heart. The mechanosensitive channel(s) responsible for regulating plasma volume remain to be determined. Here we report that the TRP channel family members TRPC1 and TRPV4 were expressed in sensory nerve endings in the atrial endocardium. Furthermore, TRPC1 and TRPV4 were coincident with the nerve ending vesicle marker synaptophysin. Calcitonin gene related peptide was exclusively confined to the myo- and epicardium of the atria. The small conductance Ca(2+)-activated K(+) channels (SK2 and SK4) were also present, however there was no relationship between SK and TRP channels. SK2 channels were expressed in nerves in the epicardium, while SK4 channels were in some regions of the endocardium but appeared to be present in epithelial cells rather than sensory endings. In conclusion, we have provided the first evidence for TRPC1 and TRPV4 channels as potential contributors to mechanosensation in the atrial volume receptors.
    Neuroscience 03/2014; 267. DOI:10.1016/j.neuroscience.2014.02.047
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    ABSTRACT: LINC (Linker of Nucleoskeleton and Cytoskeleton) complex is an evolutionary conserved structure that spans the entire nuclear envelope (NE), and integrates the nuclear interior with the cytoskeleton, in order to support a diverse array of fundamental biological processes. A key component of the LINC complex are nesprins (Nuclear Envelope SPectrin Repeat proteINS) that were initially described as large integral NE proteins. However, nesprin genes are complex and generate many variants, which occupy various sub-cellular compartments suggesting additional functions. Hence, the potential involvement of nesprins in disease has expanded immensely on what we already know. That is, nesprins are implicated in diseases such as: cancer, myopathies, arthrogryposis, neurological disorders and hearing loss. Here we review nesprins by providing an in depth account of their structure, molecular interactions and cellular functions with relevance to their potential roles in disease. Specifically, we speculate about possible pathomechanisms underlying nesprin-associated diseases.
    Seminars in Cell and Developmental Biology 12/2013; 29. DOI:10.1016/j.semcdb.2013.12.010
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    ABSTRACT: A pathological feature of heart failure (HF) is abnormal control of the sympathetic nervous system. The paraventricular nucleus of the hypothalamus (PVN) is one of the most important central sites involved in regulating sympathetic tone and is in part responsible for the dysregulation of the sympathetic nervous system evident in HF. Generation of sympathetic tone in response to fluctuations in cardiovascular regulation utilises discrete anatomical pathways and neurochemical modulators. Direct and indirect projections from the PVN-preautonomic neurons innervate the sympathetic preganglionic neurons in the spinal cord, which in turn innervate sympathetic ganglia that give rise to the sympathetic nerves. Preautonomic neurons of the PVN themselves receive an afferent input arising from the nucleus tractus solitarii (NTS) and viscerosensory receptors convey cardiovascular fluctuations to the NTS. The PVN contains excitatory and inhibitory neurons, whose balance determines the sympathetic tone. In non-pathological conditions the tonic inhibition of the PVN-preautonomic neurons is mediated by γ-aminobutyric acid (GABA) and nitric oxide (NO) releasing neurons. In HF, the preautonomic neurons are disinhibited by the actions of the excitatory neurotransmitters glutamate and angiotensin II (ANGII) leading to increased sympathetic activity. A key feature of the disinhibition is a reduction in the bioavailability of NO as a consequence of disrupted CAPON and PIN signalling mechanisms within the neuron. Another critical feature that contributes to increased neuronal excitation within the PVN is the production of proinflammatory cytokines immediately following a myocardial infarction, the activation of the ANGII-type-1 receptor and the production of reactive oxygen species. By examining the changes associated with the sympathetic nervous system pathway we will progress our understanding of sympathetic regulation in HF, identify gaps in our knowledge and suggest new therapeutic strategies.
    Experimental physiology 12/2013; 99(2). DOI:10.1113/expphysiol.2013.072678
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    ABSTRACT: We have recently developed a range of synthetic retinoid analogues which include the compounds EC23 and EC19. They are stable on exposure to light and are predicted to be resistant to the normal metabolic processes involved in the inactivation of retinoids in vivo. Based on the position of the terminal carboxylic acid groups in the compounds we suggest that EC23 is a structural analogue of all-trans retinoic acid (ATRA), and EC19 is an analogue of 13-cis retinoic acid. Their effects on the differentiation of pluripotent stem cells has been previously described in vitro and are consistent with this hypothesis. We present herein the first description of the effects of these molecules in vivo. Retinoids were applied to the anterior limb buds of chicken embryos in ovo via ion-exchange beads. We found that retinoid EC23 produces effects on the wing digits similar to ATRA, but does so at two orders of magnitude lower concentration. When larger quantities of EC23 are applied, a novel phenotype is obtained involving production of multiple digit 1s on the anterior limb. This corresponds to differential effects of ATRA and EC23 on sonic hedgehog (shh) expression in the developing limb bud. With EC23 application we also find digit 1 phenotypes similar to thumb duplications described in the clinical literature. EC23 and ATRA are shown to have effects on the entire proximal-distal axis of the limb, including hitherto undescribed effects on the scapula. This includes suppression of expression of the scapula marker Pax1. EC23 also produces effects similar to those of ATRA on the developing face, producing reductions of the upper beak at concentrations two orders of magnitude lower than ATRA. In contrast, EC19, which is structurally very similar to EC23, has novel, less severe effects on the face and rarely alters limb development. EC19 and ATRA are effective at similar concentrations. These results further demonstrate the ability of retinoids to influence embryonic development. Moreover, EC23 represents a useful new tool to investigate developmental processes and probe the mechanisms underlying congenital abnormalities in vertebrates including man.
    Journal of Anatomy 12/2013; 224(4). DOI:10.1111/joa.12147
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    ABSTRACT: This study describes the development of a functional porous polymer for use as a scaffold to support 3D hepatocyte culture. A high internal phase emulsion (HIPE) is prepared containing the monomers styrene (STY), divinylbenzene (DVB), and 2-ethylhexyl acrylate (EHA) in the external oil phase and the monomer acrylic acid (Aa) in the internal aqueous phase. Upon thermal polymerization with azobisisobutyronitrile (AIBN), the resulting porous polymer (polyHIPE) is found to have an open-cell morphology and a porosity of 89%, both suitable characteristics for 3D cell scaffold applications. X-ray photo-electron spectroscopy reveals that the polyHIPE surface contained 7.5% carboxylic acid functionality, providing a useful substrate for subsequent surface modifications and bio-conjugations. Initial bio-compatibility assessments with human hepatocytes show that the acid functionality does not have any detrimental effect on cell adhesion. It is therefore believed that this material can be a useful precursor scaffold towards 3D substrates that offer tailored surface functionality for enhanced cell adhesion.
    Macromolecular Rapid Communications 12/2013; 34(23-24). DOI:10.1002/marc.201300709
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    ABSTRACT: Artificial epidermis can be reconstituted in vitro by seeding primary epidermal cells (keratinocytes) onto a supportive substrate and then growing the developing skin equivalent at the air-liquid interface. In vitro skin models are widely used to study skin biology and for industrial drug and cosmetic testing. Here, we describe updated methods for growing 3-dimensional skin equivalents using de-vitalized, de-epidermalized dermis (DED) substrates including methods for DED substrate preparation, cell seeding, growth conditions, and fixation procedures.
    Methods in molecular biology (Clifton, N.J.) 11/2013; DOI:10.1007/7651_2013_47
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