Chick PTPσ regulates the targeting of retinal axons within the optic tectum

Chick PTP (cPTP), also known as CRYP, is a receptor-like protein tyrosine phosphatase found on axons and growth cones. Putative ligands for cPTP are distributed within basement membranes and on glial end feet of the retina, optic nerve, and optic tectum, suggesting that cPTP signaling is occurring along the whole retinotectal pathway. We have shown previously that cPTP plays a role in supporting the retinal phase of axon outgrowth. Here we have now addressed the role of cPTP within retinal axons as they undergo growth and topographic targeting in the optic tectum. With the use of retroviruses, a secretable cPTP ectodomain was ectopically expressed in ovo in the developing chick optic tectum, with the aim of directly disrupting the function of endogenous cPTP. In ovo, the secreted ectodomains accumulated at tectal sites in which cPTP ligands are also specifically found, suggesting that they are binding to these endogenous ligands. Anterograde labeling of retinal axons entering these optic tecta revealed abnormal axonal phenotypes. These included the premature stalling and arborization of fibers,excessive pretectal arbor formation, and diffuse termination zones. Most of the defects were rostral of the predicted termination zone, indicating that cPTP function is necessary for sustaining the growth of retinal axons over the optic tectum and for directing axons to their correct sites of termination. This demonstrates that regulation of cPTP signaling in retinal axons is required for their topographic mapping, the first evidence of this function for a receptor-like protein tyrosine phosphatase in the retinotectal projection

RhoE Is Regulated by Cyclic AMP and Promotes Fusion of Human BeWo Choriocarcinoma Cells

Fusion of placental villous cytotrophoblasts with the overlying syncytiotrophoblast is essential for the maintenance of successful pregnancy, and disturbances in this process have been implicated in pathological conditions such as pre-eclampsia and intra-uterine growth retardation. In this study we examined the role of the Rho GTPase family member RhoE in trophoblast differentiation and fusion using the BeWo choriocarcinoma cell line, a model of villous cytotrophoblast fusion. Treatment of BeWo cells with the cell permeable cyclic AMP analogue dibutyryl cyclic AMP (dbcAMP) resulted in a strong upregulation of RhoE at 24h, coinciding with the onset of fusion. Using the protein kinase A (PKA)-specific cAMP analogue N6-phenyl-cAMP, and a specific inhibitor of PKA (14–22 amide, PKI), we found that upregulation of RhoE by cAMP was mediated through activation of PKA signalling. Silencing of RhoE expression by RNA interference resulted in a significant decrease in dbcAMP-induced fusion. However, expression of differentiation markers human chorionic gonadotrophin and placental alkaline phosphatase was unaffected by RhoE silencing. Finally, we found that RhoE upregulation by dbcAMP was significantly reduced under hypoxic conditions in which cell fusion is impaired. These results show that induction of RhoE by cAMP is mediated through PKA and promotes BeWo cell fusion but has no effect on functional differentiation, supporting evidence that these two processes may be controlled by separate or diverging pathways

Tyrosine-Phosphorylated Caveolin-1 Blocks Bacterial Uptake by Inducing Vav2-RhoA-Mediated Cytoskeletal Rearrangements

During the early stages of infection, Neisseria gonorrhoeae triggers a phosphotyrosine-dependent Cav1-Vav2-RhoA signaling cascade that promotes the pathogen's extracellular state

Protein tyrosine phosphatases expression during development of mouse superior colliculus

Protein tyrosine phosphatases (PTPs) are key regulators of different processes during development of the central nervous system. However, expression patterns and potential roles of PTPs in the developing superior colliculus remain poorly investigated. In this study, a degenerate primer-based reverse transcription-polymerase chain reaction (RT-PCR) approach was used to isolate seven different intracellular PTPs and nine different receptor-type PTPs (RPTPs) from embryonic E15 mouse superior colliculus. Subsequently, the expression patterns of 11 PTPs (TC-PTP, PTP1C, PTP1D, PTP-MEG2, PTP-PEST, RPTPJ, RPTPε, RPTPRR, RPTPσ, RPTPκ and RPTPγ) were further analyzed in detail in superior colliculus from embryonic E13 to postnatal P20 stages by quantitative real-time RT-PCR, Western blotting and immunohistochemistry. Each of the 11 PTPs exhibits distinct spatiotemporal regulation of mRNAs and proteins in the developing superior colliculus suggesting their versatile roles in genesis of neuronal and glial cells and retinocollicular topographic mapping. At E13, additional double-immunohistochemical analysis revealed the expression of PTPs in collicular nestin-positive neural progenitor cells and RC-2-immunoreactive radial glia cells, indicating the potential functional importance of PTPs in neurogenesis and gliogenesis

Predicting small molecule fluorescent probe localization in living cells using QSAR modeling. 2. Specifying probe, protocol and cell factors; selecting QSAR models; predicting entry and localization

We describe the practical issues and the methodological procedures that must be carried out to construct and use QSAR models for predicting localization of probes in single cells. We address first the determination of probe factors starting with a consideration of the chemical nature of probe molecules present. What is their identity? Do new compounds arise in incubation media or intracellularly? For each probe, how many distinct chemical species are present? For each probe species, the derivation of the following numerical structure parameters, or descriptors, is set out with worked examples of electric charge and acid/base strength (Z and pKa); hydrophilicity/lipophilicity (log P); amphiphilicity (AI and HGH); conjugated bond number and largest conjugated fragment (CBN and LCF); width and length (W and L); and molecular and ionic weights, head group size and substituent bulk (MW, IW, HGS and SB). Next, protocol factors are specified by focusing separately on the mode of introduction of the probe to the cells, other application phenomena, and factors that influence directly observations of outcomes. Cell factors then are specified by considering separately structural and functional aspects. The next step is to select appropriate QSAR models and to integrate probe, protocol and cell factors to predict the interactions of the probe with the cell. Finally, we use an extended case example to explore the intracellular localization of certain photodynamic therapy dyes to illustrate these procedures

Entry into the legal professions The Law Student Cohort Study years 1 and 2

SIGLEAvailable from British Library Document Supply Centre-DSC:7771.850(15) / BLDSC - British Library Document Supply CentreGBUnited Kingdo

Uptake and localization mechanisms of fluorescent and colored lipid probes: part 3. protocols for predicting intracellular localization of lipid probes using QSAR models

We provide detailed protocols for applying the QSAR decision-rule models described in Part 2 of this paper. These procedures permit prediction of the intracellular localization of fluorescent probes or of any small molecular xenobiotic whether fluorescent or not. Also included is a set of notes that give practical advice on various possible problems and limitations of the methods, together with a flow chart that provides a graphical algorithmic summary of the QSAR models

Uptake and localisation of small-molecule fluorescent probes in living cells: a critical appraisal of QSAR models and a case study concerning probes for DNA and RNA

Small-molecule fluorochromes are used in biology and medicine to generate informative microscopic and macroscopic images, permitting identification of cell structures, measurement of physiological/physicochemical properties, assessment of biological functions and assay of chemical components. Modes of uptake and precise intracellular localisation of a probe are typically significant factors in its successful application. These processes and localisations can be predicted using quantitative structure activity relations (QSAR) models, which correlate aspects of the physicochemical properties of the probes (expressed numerically) with the uptake/localisation. Pay-offs of such modelling include better understanding and trouble-shooting of current and novel probes, and easier design of future probes (“guided synthesis”). Uptake models discussed consider adsorptive (to lipid or protein domains), phagocytic and pinocytotic endocytosis, as well as passive diffusion. Localisation models discussed include those for cytosol, endoplasmic reticulum, Golgi apparatus, lipid droplets, lysosomes, mitochondria, nucleus and plasma membrane. A case example illustrates how such QSAR modelling of probe interactions can clarify localisation and mode of binding of probes to intracellular nucleic acids of living cells, including not only eukaryotic chromatin DNA and ribosomal RNA, but also prokaryote chromosomes

Are secondary school students from the Middle East independent learners?

Several factors influence the quality of higher education, inputs such as quality of students and teachers, the curriculum and the pedagogy. The purpose of our research was to examine whether there were any differences in learner autonomy as measured by self-directed learning readiness (SDLR) between secondary school students who entered medicine with a local Bahraini school certificate and those students who entered with an international school certificate. Using a modified questionnaire we identified how elements such as self-management, desire for learning, self-control and total SDLR scores varied in relation to the student’s previous exit award: ‘A’ levels (or equivalent) or Bahrain Secondary School (BSS) certificate. BSS certificate students had a significantly lower mean standardised desire for learning score (63.5) compared to those entering with ‘A’ levels or equivalent (73.6; p=0.003). BSS certificate students also had a significantly lower mean total self-directed learning readiness score (192.3) compared to those students with the ‘A’ levels and equivalent (214.5; p=0.015). When we controlled for all the other factors, secondary school award certificate was the only independent predictor of self-control (standardised beta 0.4; p=0.02) and SDLR (standardised beta 0.36; p=0.043). Social shifts and changing economic workforce requirements both regionally and globally are driving an increased interest in higher education in the Middle East. Students who exit with a local secondary school certificate are finding it difficult to prepare themselves for independent learning in medical school. This poses a challenge for higher education institutions bringing a more learner autonomous type of curriculum to the Middle East

Predicting small molecule fluorescent probe localization in living cells using QSAR modeling. 1. Overview and models for probes of structure, properties and function in single cells

Small molecule fluorochromes (synonyms: biosensors, chemosensors, fluorescent probes, vital stains) are widely used to investigate the structure, composition, physicochemical properties and biological functions of living cells, tissues and organisms. Selective entry and accumulation within particular cells and cellular structures are key processes for achieving these diverse objectives. Despite the complexities, probes routinely are applied using standard protocols, often without experimenter awareness of what factors that control accumulation and localization. The mechanisms of many such selective accumulations, however, now are known. Moreover, the influence of physicochemical properties of probes on their uptake and localization often can be defined numerically, hence predicted, using quantitative structure activity relations (QSAR) models with its required numerical structure parameters (or “descriptors”). The state of the art of this approach is described. Available QSAR models are summarized for uptake into cells and localization in the cytosol, endoplasmic reticulum, generic biomembranes, Golgi apparatus, lipid droplets, lysosomes/endosomes, mitochondria, eukaryotic nuclei (histones and DNA), plasma membrane, and ribosomal RNA (cytoplasmic and nucleolar). Integration of such core models to both aid understanding and troubleshooting of current fluorescent probes and to assist the design of novel probes is outlined and illustrated using case examples. Limitations and generic problems arising with this approach and comments on application of such approaches to xenobiotics other than probes, e.g., drugs and herbicides, together with a brief note about an alternative approach to prediction, are given