Search

Progressive neurodegenerative disorders such as Parkinson Disease (PD) lack curative or long-term treatments. At the same time, the increase of the worldwide elderly population and, consequently, the extension in the prevalence of age-related diseases have promoted research interest in neurodegenerative disorders. Caenorhabditis elegans is a free-living nematode widely used as an animal model in studies of human diseases. Here we evaluated cannabidiol (CBD) as a possible neuroprotective compound in PD using the C. elegans models exposed to reserpine. Our results demonstrated that CBD reversed the reserpine-induced locomotor alterations and this response was independent of the NPR-19 receptors, an orthologous receptor for central cannabinoid receptor type 1. Morphological alterations of cephalic sensilla (CEP) dopaminergic neurons indicated that CBD also protects neurons from reserpine-induced degeneration. That is, CBD attenuates the reserpine-induced increase of worms with shrunken soma and dendrites loss, increasing the number of worms with intact CEP neurons. Finally, we found that CBD also reduced ROS formation and α-syn protein accumulation in mutant worms. Our findings collectively provide new evidence that CBD acts as neuroprotector in dopaminergic neurons, reducing neurotoxicity and α-syn accumulation highlighting its potential in the treatment of PD. [ABSTRACT FROM AUTHOR]

Context: Xanthates are organic compounds that present great interest for coordination chemistry, because they can bond in different ways to the metal ion. Thus, these compounds have several applications, being best known for their environmental application. In fact, xanthates are recognized for their application as heavy metal collector agents in aqueous environments. In view of this application, this study is aimed at showing the thermochemical and electronic parameters obtained for the reactions of substitution water molecules in the aqua zinc complexes, by xanthate ligands (n-propyl, n-butyl, and n-pentyl xanthates). In addition to their environmental application, xanthates have shown biological properties, such as anti-bacterial and anti-cancer. In recent years, xanthates have also been used in the technological area, where it participates as a precursor of sulfides for the manufacture of thin films. Our results showed complexes with distorted octahedral geometries and with negative values of enthalpy and Gibbs free energy, indicating exothermic and spontaneous processes. For all the complexes, it was observed that Zn2+ complexes have both an ionic and covalent character. However, the monosubstituted complexes showed a predominance of the ionic character. In addition, high donor-acceptor interaction energies were obtained, indicating a good superposition between the s and p orbitals involved in the Zn-S bond. Methods: This work consists in theoretical studies of Zn2+ complexes with alkyl xanthate ligands, with different structures, where optimization and normal modes calculations were performed at different DFT levels: M06L, M06-2X, wB97XD, and B3LYP/6-311++G**+LANL2TZ, with Gaussian09 program. The process of substitution of two aqua by two xanthate ligands was analyzed in stages, forming cationic and neutral complexes, in the first and second stages, respectively. In addition, electronic energy decomposition (EDA) and natural bond orbital (NBO) analysis were performed at level M06L/6-311++G**+LANL2TZ with Gamess program. [ABSTRACT FROM AUTHOR]

Photocatalysis is considered to be one of the possible routes to solve energy crises at a low cost. Graphitic-C3N4 combined with other semiconductors at the nanoscale has demonstrated a promising potential in photocatalysis application owing to the heterojunction configuration. In this study, Ag/g-C3N4/CuNb2O6-4 (mass ratio of CuNb2O6 to g-C3N4 = 1:4) composites were prepared by photo-deposition of Ag particles on the hydrothermally synthesized g-C3N4/CuNb2O6-4 composites. The microstructure, morphology, and light absorption property of the photocatalysts were characterized by X-ray diffractometry, scanning electron microscopy, transmission electron microscopy, X-ray photoelectron spectroscopy, and UV–Vis diffuse reflectance spectroscopy. The photocatalytic properties of photocatalysts and active species responsible for the degradation of RhB dye under visible light were investigated. After loading Ag particles on the g-C3N4/CuNb2O6-4 composites, the resulting Ag/g-C3N4/CuNb2O6-4 composites show a bandgap of ~ 2.31 eV and a significant increase in photocatalytic efficiency. In particular, the photocatalytic activity of 11% Ag/g-C3N4/CuNb2O6-4 composite (Ag mass fraction = 11%) is nearly seven times that of orthorhombic CuNb2O6 and 3.8 times that of g-C3N4/CNO-4 composite. The Z-scheme heterojunction has been successfully formed in the Ag/g-C3N4/CuNb2O6-4 composites and it enhances the photocatalytic performance significantly with the best degradation efficiency of 88% after 2 h. [ABSTRACT FROM AUTHOR]

Herein, a flexible light weight lead-free piezoelectric nanogenerator has been fabricated using electrospun web based on polyacrylonitrile (PAN) and copper oxide (CuO) nanorods. The influence of CuO nanorods on the piezoelectric performance of PAN nanofibrous structure (in the form of a nanocomposite) has been investigated critically. Structural analysis of the electrospun nanocomposite has been done by FTIR and XRD techniques. Morphology of the developed material has been evaluated by scanning electron microscopy (SEM), whereas thermal behaviour has been investigated by thermogravimetric analyser (TGA) and differential scanning calorimetry (DSC). Finally, piezoelectric performance of the device has been measured by a digital oscilloscope. The piezoelectric nanogenerator (PAN/0.5% CuO) has shown an output voltage of ~ 5 V, current ~ 0.172 µA and power density ~ 0.215 µW/cm2 (when pressure is applied by finger tapping). The same nanogenerator has shown ~ 1.25 V and ~ 118 nA output voltage and short circuit current, respectively by using an automatic force-imparting machine. The fabricated nanogenerator is a potent device in lieu of the conventional battery powered devices. [ABSTRACT FROM AUTHOR]

Cadmium selenide quantum dots (QDs) are fluorescent and electron-dense nanoparticles. When used as reporter of immunolabeling, this dual visibility is essential for direct comparison of its fluorescent signals on light microscopy (LM) and their ultrastructrual counterparts on electron microscopy (EM) as correlative light and electron microscopy (CLEM). To facilitate EM recognition, QDs on EM grid were gold enhanced, which increased their size and electron density. On histological sections as well, gold-enhanced QDs, used as a reporter of immunolabeling, were easily recognized on EM. Because target structures are visible on bright field microscopy, gold enhancement facilitated trimming the target structures into final EM sections. Furthermore, gold enhancement of rod-shaped QD655 on EM grid was accentuated on their tips while spherical QD565 was gold-enhanced as sphere in contrast. This EM distinction was evident on histological sections where QD565 (green fluorescence) and QD655 (red fluorescence) were used as a reporter pair for double immunolabeling. Double-labeled immuno-fluorescent images, initially captured before EM processing, are now compared with their respective immuno EM counterparts. Specific labeling of each epitope was corroborated by mutual comparison between LM and EM. Although fluoronanogold may be a candidate reporter partner with QDs for gold-enhanced, double-labeling CLEM, its limited penetration into fixed tissue hampers universal use for thick histological sections. Gold-enhancement of QD immunolabeling, now expanded to double-labeling CLEM for human brain samples, will pave the way to translate molecular events into ultrastructural morphopathogenesis in situ. [ABSTRACT FROM AUTHOR]

Background Studies have demonstrated the potential for patient feedback to inform quality care as well as a direct relationship between patient experience and clinical outcomes. Over recent years, there has been increasing use of online patient feedback platforms, however, there has been little study of the content of patient feedback relating to pharmacy and pharmacy services. Objective This study explores the content of online feedback provided by patients from across the UK in relation to their experiences of their interaction with pharmacy staff and pharmacy services. Main outcome measure Content of online patient feedback relating to pharmacy. Method Patient stories published on Care Opinion, a national online patient feedback platform, for a one-year period were searched for all content relating to patients' pharmacy experiences. A thematic and sentiment analysis was conducted on 237 patient stories. Results Patient stories related to supply, staff attitudes, services, accessibility, systems, and errors. Patient sentiment depended on pharmacy setting, but staff attitudes, services, and accessibility were generally positive across all settings. Waiting time was the most common complaint in both hospital and community pharmacies with stories relaying experiences of slow discharge, stock shortages and poor communication and collaboration between pharmacies and GP surgeries. Conclusions Online patient feedback highlighted factors important to patients when interacting with pharmacies and their staff. Medication supply was the primary topic of patient stories with waiting times and stock shortages being clear areas for improvement; however, accessibility, pharmacy services and advice were key strengths of the profession. Further research is needed to understand how online patient feedback can be used effectively to inform improvements in pharmacy services. [ABSTRACT FROM AUTHOR]

Cells, basic units of living structures and functions, build up a complicated small world, and their order and complexity resemble a small universe. The detailed understanding and elucidation of the matter transport and energy conversion mechanisms within a single cell will expand our knowledge about the origin and evolution of life, the disease mechanisms and much more. In past decades, single-cell analysis has been rapidly and significantly improved and various methodologies have been developed to reveal the complexity of mass, energy, and information within single cells. In this review, we focused on the methods developed in recent years for single-cell analysis, including electrochemical method, optical method, and mass spectrometry method. We reviewed the recent advances and representative studies in this research field, and also discussed the strengths and limitations of each method. Finally, we presented the existing technical challenges and further directions for single-cell analysis. [ABSTRACT FROM AUTHOR]

Cryo-correlative light and electron microscopy (Cryo-CLEM) is materializing as a widespread approach amalgamating the advantages of both fluorescence light microscopy (FLM) as well as three dimensional (3D) cryo-electron tomography (cryo-ET) to reveal the ultrastructure of significant target molecules with specific cellular functions. Cryo-CLEM allows imaging of cells by means of fluorescence microscopy exhibiting the location of the destined molecule at high temporal and spatial resolution while cryo-ET is employed to analyze the 3D structure at a molecular resolution in close-to-physiological condition. Present review focuses upon the practical strategies for Cryo-CLEM and recent technical developments that will assist the broad implementation of this technique to investigate and answer questions pertaining to various biological events occurring in the cell. [ABSTRACT FROM AUTHOR]

An animal's stress response requires different adaptive strategies depending on the nature and duration of the stressor. Whereas acute stressors, such as predation, induce a rapid and energy-demanding fight-or-flight response, long-term environmental stressors induce the gradual and long-lasting activation of highly conserved cytoprotective processes1–3. In animals across the evolutionary spectrum, continued activation of the fight-or-flight response weakens the animal's resistance to environmental challenges4,5. However, the molecular and cellular mechanisms that regulate the trade-off between the flight response and long-term stressors are poorly understood. Here we show that repeated induction of the flight response in Caenorhabditis elegans shortens lifespan and inhibits conserved cytoprotective mechanisms. The flight response activates neurons that release tyramine, an invertebrate analogue of adrenaline and noradrenaline. Tyramine stimulates the insulin–IGF-1 signalling (IIS) pathway and precludes the induction of stress response genes by activating an adrenergic-like receptor in the intestine. By contrast, long-term environmental stressors, such as heat or oxidative stress, reduce tyramine release and thereby allow the induction of cytoprotective genes. These findings demonstrate that a neural stress hormone supplies a state-dependent neural switch between acute flight and long-term environmental stress responses and provides mechanistic insights into how the flight response impairs cellular defence systems and accelerates ageing. The release of tyramine during the flight response in nematodes activates the DAF-2/insulin–IGF pathway to downregulate cytoprotective mechanisms and shorten lifespan. [ABSTRACT FROM AUTHOR]

At the presynaptic terminal, neurotransmitters are stored in synaptic vesicles (SVs), which are released and recycled via exo- and endocytosis. SV endocytosis is crucial for sustaining synaptic transmission by maintaining the SV pool. Many studies have shown that presynaptic dysfunction, particularly impairment of SV endocytosis, is related to neurological disorders. Notably, the presynaptic terminal is considered to be a sensitive structure because certain presynaptic dysfunctions, manifested as impaired SV endocytosis or ultrastructural changes in the presynaptic terminal, can be observed before there is a biochemical or pathological evidence of a neurological disorder. Therefore, monitoring and assessing the presynaptic function by SV endocytosis facilitates the development of early markers for neurological disorders. In this study, we reviewed the current methods for assessing and visualizing SV endocytosis at the central nerve terminal. [ABSTRACT FROM AUTHOR]

Background Errors in discharge prescriptions are problematic. When hospital pharmacists write discharge prescriptions improvements are seen in the quality and efficiency of discharge. There is limited information on the incidence of errors in pharmacists' medication orders. Objective To investigate the extent and clinical significance of errors in pharmacist-written discharge medication orders. Setting 1000-bed teaching hospital in London, UK. Method Pharmacists in this London hospital routinely write discharge medication orders as part of the clinical pharmacy service. Convenient days, based on researcher availability, between October 2013 and January 2014 were selected. Pre-registration pharmacists reviewed all discharge medication orders written by pharmacists on these days and identified discrepancies between the medication history, inpatient chart, patient records and discharge summary. A senior clinical pharmacist confirmed the presence of an error. Each error was assigned a potential clinical significance rating (based on the NCCMERP scale) by a physician and an independent senior clinical pharmacist, working separately. Main outcome measure Incidence of errors in pharmacist-written discharge medication orders. Results 509 prescriptions, written by 51 pharmacists, containing 4258 discharge medication orders were assessed (8.4 orders per prescription). Ten prescriptions (2%), contained a total of ten erroneous orders (order error rate-0.2%). The pharmacist considered that one error had the potential to cause temporary harm (0.02% of all orders). The physician did not rate any of the errors with the potential to cause harm. Conclusion The incidence of errors in pharmacists' discharge medication orders was low. The quality, safety and policy implications of pharmacists routinely writing discharge medication orders should be further explored. [ABSTRACT FROM AUTHOR]

We studied the structural formation process of polyacrylonitrile (PAN) gelation by a wide-spatial scale. In micron-scale structure, light transmittance and gelation time were evaluated. The presence of water in the mixed solvent accelerated the phase separation and gelation because of the poor solubility of PAN in water, while in the case of low water concentration, transparent gelation process could be observed. Further, we conducted in situ small-angle neutron scattering measurements to clarify the gelation processes of 10 wt% PAN solutions with various water contents with new-type spectrometer in MLF, J-PARC, Japan. The nanometer-scaled inhomogeneities in the polymer chains develop immediately after quenching. In the opaque-gel case, we observed the nanometer-scaled correlation of crystalline cross-linking points and smooth boundary of two micron-scaled phases due to liquid-liquid phase separation. Cross-linking points begin to emerge and grow just after quenching. The correlation length is independent of the annealing time, then the cross-linking point exists locally and micro-gelation occurs during phase separation. By contrast, in the transparent-gel case, we did not observe scattering from the surfaces of the cross-linking points; we were able to measure only the correlation length. The correlation length was found to increase throughout the gelation process, and the entanglements of the polymer chains play a role similar to that of cross-linking points. [ABSTRACT FROM AUTHOR]

Background: Traditionally, hospital pharmacists' roles have been associated with dispensing medications prescribed by doctors and offering advice about medicines to patients and other healthcare professionals. In England, significant changes in the structure of hospital pharmacy practice began in the 1970s and currently hospital pharmacists are undertaking a number of advanced roles including prescribing.Objective: This study investigated the facilitators to change in hospital pharmacy practice in England in order to identify lessons that might assist in the potential changes needed in other countries for extended clinical roles.Setting: The study was conducted in England.Methods: A qualitative study using semi-structured interviews was conducted with 28 participants, comprising 22 pharmacists and 6 pharmacy technicians from England. They were recruited through a snowball sampling technique. Transcribed interviews were entered into the QSR NVivo 10 software for data management and analysed thematically. Main outcome measure Pharmacists and pharmacy technicians' perception of the facilitators to hospital pharmacy practice change in England.Result: Three major themes emerged from this study: drivers for change, strategies for change and efficiency. Many of the drivers identified were linked to changes in the structure of hospital pharmacy including education and training; specialisation in practice and career structure. Strategies employed to achieve practice change included broadening the role of pharmacy technicians in order to free-up pharmacists' time; seizing opportunities for extended roles; developing a relationship with the medical profession and professional leadership influence. Participants perceived that the development of pharmacists' clinical roles have resulted in a more efficient healthcare provision where patients were offered seamless services.Conclusion: Changes in the professional structure of pharmacy including education and training, specialisation, career structure and the roles of pharmacy technicians could benefit the development of pharmacists' clinical roles in other countries. [ABSTRACT FROM AUTHOR]

Despite intensive research, it is still unclear how an immediate and profound acceleration of exocytosis is triggered by appropriate Ca-stimuli in presynaptic terminals. This is due to the fact that the molecular mechanisms of 'docking' and 'priming' reactions, which set up secretory vesicles to fuse at millisecond time scale, are extremely hard to study. Yet, driven by a fruitful combination of in vitro and in vivo analyses, our mechanistic understanding of Ca-triggered vesicle fusion has certainly advanced in the past few years. In this review, we aim to highlight recent progress and emerging views on the molecular mechanisms, by which constitutively forming SNAREpins are organized in functional, tightly regulated units for synchronized release. In particular, we will focus on the role of the small regulatory factor complexin whose function in Ca-dependent exocytosis has been controversially discussed for more than a decade. Special emphasis will also be laid on the functional relationship of complexin and synaptotagmin, as both proteins possibly act as allies and/or antagonists to govern SNARE-mediated exocytosis. [ABSTRACT FROM AUTHOR]

Ultrastructural examination of cells and tissues by electron microscopy (EM) yields detailed information on subcellular structures. However, EM is typically restricted to small fields of view at high magnification; this makes quantifying events in multiple large-area sample sections extremely difficult. Even when combining light microscopy (LM) with EM (correlated LM and EM: CLEM) to find areas of interest, the labeling of molecules is still a challenge. We present a new genetically encoded probe for CLEM, named 'FLIPPER', which facilitates quantitative analysis of ultrastructural features in cells. FLIPPER consists of a fluorescent protein (cyan, green, orange, or red) for LM visualization, fused to a peroxidase allowing visualization of targets at the EM level. The use of FLIPPER is straightforward and because the module is completely genetically encoded, cells can be optimally prepared for EM examination. We use FLIPPER to quantify cellular morphology at the EM level in cells expressing a normal and disease-causing point-mutant cell-surface protein called EpCAM (epithelial cell adhesion molecule). The mutant protein is retained in the endoplasmic reticulum (ER) and could therefore alter ER function and morphology. To reveal possible ER alterations, cells were co-transfected with color-coded full-length or mutant EpCAM and a FLIPPER targeted to the ER. CLEM examination of the mixed cell population allowed color-based cell identification, followed by an unbiased quantitative analysis of the ER ultrastructure by EM. Thus, FLIPPER combines bright fluorescent proteins optimized for live imaging with high sensitivity for EM labeling, thereby representing a promising tool for CLEM. [ABSTRACT FROM AUTHOR]

Background The last decade has seen a drive towards non-medical prescribing in the United Kingdom (UK). However, there is a dearth of any published literature on applying the principles of service redesign to support pharmacist prescribing in any sphere of practice. Objective To develop consensus guidance to facilitate service redesign around pharmacist prescribing. Setting UK hospital practice. Methods The Delphi technique was used to measure consensus of a panel of expert opinion holders in Scotland. Individuals with key strategic and operational roles in implementing initiatives of pharmacy practice and medicines management were recruited as experts. An electronic questionnaire consisting of 30 statements related to pharmacist prescribing service redesign was developed. These were presented as five-point Likert scales with illustrative quotes. Main outcome measures Consensus, defined as 70 % of panel members agreeing (ranked strongly agree/agree) with each statement. Results Responses were obtained from 35/40 (87.5 %) experts in round one and 29 (72.5 %) in round two. Consensus in round one was achieved for 27/30 of statements relating to aspects of generic 'service development' (e.g. succession planning, multidisciplinary working, quality evaluation, practice development and outcome measures) and 'pharmacist prescribing role development' (e.g. education and future orientation of service). Issues of disagreement were around targeting of pharmacist prescribing to clinical specialities and financial remuneration for prescribing in the hospital setting. Conclusion Consensus guidance has been developed to facilitate service redesign around hospital pharmacist prescribing. [ABSTRACT FROM AUTHOR]

In these last two decades , fluorescent proteins (FPs) have become highly valued imaging tools for cell biology, owing to their compatibility with living samples, their low levels of invasiveness and the possibility to specifically fuse them to a variety of proteins of interest. Remarkably, the recent development of phototransformable fluorescent proteins (PTFPs) has made it possible to conceive optical imaging experiments that were unimaginable only a few years ago. For example, it is nowadays possible to monitor intra- or intercellular trafficking, to optically individualize single cells in tissues or to observe single molecules in live cells. The tagging specificity brought by these genetically encoded highlighters leads to constant progress in the engineering of increasingly powerful, versatile and non-cytotoxic FPs. This review is focused on the recent developments of PTFPs and highlights their contribution to studies within cells, tissues and even living organisms. The aspects of single-molecule localization microscopy, intracellular tracking of photoactivated molecules, applications of PTFPs in biotechnology/optobiology and complementarities between PTFPs and other microscopy techniques are particularly discussed. [ABSTRACT FROM AUTHOR]

The high toxicity of the seven serotypes of botulinum neurotoxins (BoNT/A to G), together with their specificity and reversibility, includes them in the list A of potential bioterrorism weapons and, at the same time, among the therapeutics of choice for a variety of human syndromes. They invade nerve terminals and cleave specifically the three proteins which form the heterotrimeric SNAP REceptors (SNARE) complex that mediates neurotransmitter release. The BoNT-induced cleavage of the SNARE proteins explains by itself the paralysing activity of the BoNTs because the truncated proteins cannot form the SNARE complex. However, in the case of BoNT/A, the most widely used toxin in therapy, additional factors come into play as it only removes a few residues from the synaptosomal associate protein of 25 kDa C-terminus and this results in a long duration of action. To explain these facts and other experimental data, we present here a model for the assembly of the neuroexocytosis apparatus in which Synaptotagmin and Complexin first assist the zippering of the SNARE complex, and then stabilize and clamp an octameric radial assembly of the SNARE complexes. [ABSTRACT FROM AUTHOR]

Background Under UK legislation, suitably qualified non-medical professionals can practise as prescribers. Few studies have explored the views of the general public towards non-medical prescribing. Objective The aim was to explore the views of the Scottish general public on non-medical prescribing. Setting General community in Scotland. Method A pre-piloted survey was mailed to a random sample of 5,000 members of the general public in Scotland. In addition to the items on awareness of and attitudes towards non-medical prescribing, respondents were asked to 'give any other comments, issues or concerns you have in relation to health professionals other than doctors prescribing.' Responses were subjected to content analysis. Main outcome measures Key themes identified from content analysis. Results The overall questionnaire response rate was 37.1 % (n = 1,855) of which 27.2 % (n = 505) provided comments. Most were directly related to pharmacist prescribing (n = 312) while others referred to non-medical prescribers generically (n = 172) or other healthcare professionals (n = 79). Nine themes were identified: perception of knowledge and training; support for a limited range of non-medical prescribing; access to medical records; motivation and convenience; confidence, faith and trust; privacy and confidentiality; risks, controls and continuity of care; supervision and conflict of interest; communication and cooperation. Conclusions The findings identify support for non-medical prescribing but indicate the need for non-medical prescribers to engage more with the general public. The comments also provide insight into the challenges for non-medical prescribers, as they strive to fulfil their extended healthcare roles. [ABSTRACT FROM AUTHOR]

Fear, a reaction to a threatening situation, is a broadly adaptive feature crucial to the survival and reproductive fitness of individual organisms. By contrast, anxiety is an inappropriate behavioral response often to a perceived, not real, threat. Functional imaging, biochemical analysis, and lesion studies with humans have identified the HPA axis and the amygdala as key neuroanatomical regions driving both fear and anxiety. Abnormalities in these biological systems lead to misregulated fear and anxiety behaviors such as panic attacks and post-traumatic stress disorders. These behaviors are often treated by increasing serotonin levels at synapses, suggesting a role for serotonin signaling in ameliorating both fear and anxiety. Interestingly, serotonin signaling is highly conserved between mammals and invertebrates. We propose that genetically tractable invertebrate models organisms, such as Drosophila melanogaster and Caenorhabditis elegans, are ideally suited to unravel the complexity of the serotonin signaling pathways. These model systems possess well-defined neuroanatomies and robust serotonin-mediated behavior and should reveal insights into how serotonin can modulate human cognitive functions. [ABSTRACT FROM AUTHOR]

Monoamines and neuropeptides interact to modulate key behaviors in most organisms. This review is focused on the interaction between octopamine (OA) and an array of neuropeptides in the inhibition of a simple, sensory-mediated aversive behavior in the C. elegans model system and describes the role of monoamines in the activation of global peptidergic signaling cascades. OA has been often considered the invertebrate counterpart of norepinephrine, and the review also highlights the similarities between OA inhibition in C. elegans and the noradrenergic modulation of pain in higher organisms. [ABSTRACT FROM AUTHOR]

Pharmacological and genetic tools targeting the 5-hydroxytryptamine (5-HT)7 receptor in preclinical animal models have implicated this receptor in diverse (patho)physiological processes of the central nervous system (CNS). Some data obtained with 5-HT7 receptor knockout mice, selective antagonists, and, to a lesser extent, agonists, however, are quite contradictory. In this review, we not only discuss in detail the role of the 5-HT7 receptor in the CNS but also propose some hypothetical models, which could explain the observed inconsistencies. These models are based on two novel concepts within the field of G protein-coupled receptors (GPCR), namely biphasic signaling and G protein-independent signaling, which both have been shown to be mediated by GPCR dimerization. This led us to suggest that the 5-HT7 receptor could reside in different dimeric contexts and initiate different signaling pathways, depending on the neuronal circuitry and/or brain region. In conclusion, we highlight GPCR dimerization and G protein-independent signaling as two promising future directions in 5-HT7 receptor research, which ultimately might lead to the development of more efficient dimer- and/or pathway-specific therapeutics. [ABSTRACT FROM AUTHOR]

We report the ac conductivity and relaxation behavior analysis for a heterogeneous polymer-clay nanocomposite (PNC) having composition (polyacrylonitrile)LiCFSO + x wt.% dodecylamine modified montmorillonite. Charge transport behavior in an ionically conducting PNC has been analyzed systematically and correlated with the macroscopic parameters like polymer glass transition temperature and available free mobile charge carriers. Intercalation of cation coordinated polymer into the nanometric clay channels has been confirmed by high-resolution transmission electron microscopy. The electrical properties of the intercalated PNC films have been studied using complex impedance/admittance spectroscopy. Excellent correlation of relaxation behavior with polymer glass transition temperature ( T) confirmed the objectives of the work. An analysis of dielectric relaxation indicates that PNC films are lossy when compared with polymer-salt film. This result is a direct outcome of faster ion dynamics leading to strong electrode polarization effect due to the accumulation of charge carriers at the interface. [ABSTRACT FROM AUTHOR]

This review considers the factors involved in the regulation of feeding and metabolism in response to food deprivation using Caenorhabditis elegans as a model organism. Some of the sensory neurons and interneurons involved in food intake are described, together with an overview of pharyngeal pumping. A number of chemical transmitters control feeding in C. elegans including 5-hydroxytryptamine (5-HT, serotonin), acetylcholine, glutamate, dopamine, octopamine, and tyramine. The roles of these transmitters are modified by neuropeptides, including FMRFamide-like peptides (FLPs), neuropeptide-like protein (NLPs), and insulin-like peptides. The precise effects of many of these neuropeptides have yet to be elucidated but increasingly they are being shown to play a role in feeding and metabolism in C. elegans. The regulation of fat stores is complex and appears to involve the expression of a large number of genes, many with mammalian homologues, suggesting that fat regulatory signalling is conserved across phyla. Finally, a brief comparison is made between C. elegans and mammals where for both, despite their evolutionary distance, classical transmitters and neuropeptides have anorectic or orexigenic properties. Thus, there is a rationale to support the argument that an understanding of the molecular and genetic basis of feeding and fat regulation in C. elegans may contribute to efforts aimed at the identification of targets for the treatment of conditions associated with abnormal metabolism and obesity. [ABSTRACT FROM AUTHOR]

The ability to safely control transgene expression from viral vectors is a long-term goal in the gene therapy field. We have previously reported tight regulation of GFP expression in rat brain using a self-regulating tet-off rAAV vector. The immune responses against tet regulatory elements observed by other groups in nonhuman primates after intramuscular injection of tet-on encoding vectors raise concerns about the clinical value of tet-regulated vectors. However, previous studies have not examined immune responses following injection of AAV vectors into brain. Therefore, rat striatum was injected with tet-off rAAV harboring a therapeutic gene for Parkinson's disease, either hAADC or hGDNF. The expression of each gene was tightly controlled by the tet-off regulatory system. Using an ELISA developed with purified GST–tTA protein, no detectable immunogenicity against tTA was observed in sera of rats that received an intrastriatal injection of either vector. In contrast, sera from rats intradermally injected with an adenovirus containing either tTA or rtTA, as positive controls, had readily detectable antibodies. These observations suggest that tet-off rAAV vectors do not elicit an immune response when injected into rat brain and that these may offer safer vectors for Parkinson's disease than vectors with constitutive expression. [ABSTRACT FROM AUTHOR]

This paper investigates the effect of epinastine, a selective octopamine antagonist in invertebrates, in Caenorhabditis elegans. Specifically, its ability to block the inhibitory action of octopamine on C. elegans-isolated pharynx was assayed. Isolated pharynxes were stimulated to pump by the addition of 500 nM 5-hydroxytryptamine (5-HT) (113 ± 2 per 30 s, n = 15). Octopamine inhibited the 5-HT-induced pumping in a concentration-dependent manner (threshold 1-5 μM) with a 61 ± 11% inhibition with 50 μM ( n = 5). Epinastine (0.1 μM) antagonized the inhibitory response to octopamine ( P < 0.001; n = 15). Tyramine also inhibited pharyngeal pumping induced by 5-HT but was less potent than octopamine. Tyramine, 50 μM to 1 mM, gave a transient inhibition e.g. of 40 ± 5% at 50 μM ( n = 5). A higher (10 μM) concentration of epinastine was required to block the tryamine response compared with octopamine. It is concluded that epinastine selectively antagonizes the effect of octopamine on C. elegans pharynx. Further studies are required to test its selectivity for octopamine in other tissues and other nematodes. [ABSTRACT FROM AUTHOR]

In recent years, the development of powerful viral gene transfer techniques has greatly facilitated the study of gene function. This review summarises some of the viral delivery systems routinely used to mediate gene transfer into cell lines, primary cell cultures and in whole animal models. The systems described were originally discussed at a 1-day European Tissue Culture Society (ETCS-UK) workshop that was held at University College London on 1st April 2009. Recombinant-deficient viral vectors (viruses that are no longer able to replicate) are used to transduce dividing and post-mitotic cells, and they have been optimised to mediate regulatable, powerful, long-term and cell-specific expression. Hence, viral systems have become very widely used, especially in the field of neurobiology. This review introduces the main categories of viral vectors, focusing on their initial development and highlighting modifications and improvements made since their introduction. In particular, the use of specific promoters to restrict expression, translational enhancers and regulatory elements to boost expression from a single virion and the development of regulatable systems is described. [ABSTRACT FROM AUTHOR]

Abstract  A new complex of the bis(O,O′-di(2-phenylethyl)dithiophosphato) ligand, namely, Ni[S2P(OCH2CH2Ph)2]2 was synthesized and characterized by spectroscopic methods, magnetic measurements, elemental analysis and single-crystal X-ray diffraction. In the crystal, the (PhCH2CH2O)2PSS− serves as a bidentate ligand, coordinating to the nickel atom with its two sulphur atoms. The nickel atom lies on a centre of symmetry and forms two independent Ni–S bonds [2.2228(13) and 2.2202 (11)Å] in a four-coordinate plane. The hydrolysis of p-nitrophenyl picolinate (PNPP) promoted by the complex was investigated by UV–vis spectrophotometry. The results show that this complex is a very efficient catalyst for promoting the hydrolysis of PNPP. A mechanism for the hydrolysis is proposed. [ABSTRACT FROM AUTHOR]

Abstract   Aim The aim of this study was to explore patients’ perspectives and experiences of pharmacist supplementary prescribing (SP) in Scotland. Method A survey in primary and secondary care in Scotland. Pharmacist supplementary prescribers (n = 10) were purposively selected across Scotland. All pharmacists distributed questionnaires to 20 consecutive patients as they attended appointments during October to December 2006. Reminders were mailed to all 20 patients by each pharmacist 2 weeks after initial distribution. Main outcome measures The questionnaire contained items on: attitudes towards pharmacist SP derived from earlier qualitative research; consultation satisfaction derived from a validated scale developed initially for general practitioners, with the term ‘doctor’ being replaced by ‘pharmacist prescriber’; and demographics. Closed and Likert scales were used as response options. Results One pharmacist withdrew. The patient response rate was 57.2% (103/180). The median age was 67 years (interquartile range 56.5–73 years), with 53.4% being female. Most (76, 73.8%) consulted with the pharmacist in a general practice setting. Patients reported positive consultation experiences with 89.3% agreeing/strongly agreeing that they were satisfied with the consultation, 78.7% thought the pharmacist told them everything about their treatment and 72.9% felt the pharmacist was interested in them as a person. Most patients were positive in their attitudes, agreeing that they would recommend a pharmacist prescriber to others and that they had trust in the pharmacist. However, 65% would prefer to consult a doctor. Conclusion Most patient respondents were satisfied with, and had a positive attitude towards, pharmacist prescribing consultations. However, most patients would still elect to see a doctor given the choice. [ABSTRACT FROM AUTHOR]

Abstract   Objectives To design a care pathway and referral documentation system to support dyspepsia management in the community pharmacy according to national guidelines. To explore attitudes of community pharmacists and general practitioners (GPs) towards the proposed model of care. Setting Focus group of four community pharmacists from one locality and one-to-one interviews with five GPs in each of their surgeries in different city localities. Method Literature review; design of a care pathway and referral system; qualitative analysis of focus group discussion and one-to-one interviews. Main outcome measure Modified design of documentation system; attitudes and barriers towards an extended role for pharmacists in a community pharmacy based model of individual care of patients with dyspepsia. Results The care pathway and documentation system was modified to make it simple to use and to assure appropriate referral and reporting to GPs. All participants agreed that the SIGN guideline for Dyspepsia and local dyspepsia guidelines provide an opportunity to extend the role of pharmacists to prescribe therapeutic doses of H2 receptor antagonists, test for Helicobacter pylori in the pharmacy and prescribe eradication therapy. Perceived advantages to patients included better convenience and reduced waiting times for H. pylori testing. Current barriers to this model identified were: medico-legal issues, budgetary issues, limited access to patient data, lack of privacy to talk to patients, inadequate time and remuneration, no patient registration, patients describing vague symptoms and patient confidence in pharmacists. Conclusion The proposed model of care and documentation system for community pharmacy based dyspepsia management generated a favourable response and design contributions from community pharmacists and GPs. Barriers were identified which have to be overcome prior to implementation and evaluation of the model. [ABSTRACT FROM AUTHOR]

Abstract  The feeding activity of the nematode Caenorhabditis elegans is regulated by an anatomically well-defined network of 20 enteric neurones that employs small molecule and neuropeptidergic signalling. Two of the most potent excitatory agents are 5-HT and the neuropeptide FLP17A. Here we have examined the role of cAMP in modulating their excitatory actions by pharmacological manipulation of the level of cAMP. Application of the membrane permeable cAMP analogue, dibutyryl-cAMP (1 μM), enhanced the excitatory response to both FLP17A and 5-HT. Furthermore, the adenylyl cyclase activator, forskolin (50 nM), significantly enhanced the excitatory response to both FLP17A and 5-HT. The phosphodiesterase inhibitor, ibudilast (10 μM), enhanced the excitatory response to FLP17A. The protein kinase inhibitor, H-9 dihydrochloride (10 μM) significantly reduced the excitatory response to 5-HT. H-9 dihydrochloride also had a direct effect on pharyngeal activity. The effect of FLP17A and 5-HT on two mutants, egl-8 (loss-of-function phospholipase-Cβ) and egl-30 (loss-of-function Gαq) was also investigated. Both these mutants have a lower pharyngeal pumping rate than wild-type which has to be considered when interpreting the effects of these mutations on the excitatory responses to FLP17A and 5HT. However, even taking into consideration the lower basal activity of these mutants, it is clear that the percentage increase in pharyngeal pumping rate induced by FLP17A is greatly reduced in both mutants compared to wild-type. In the case of 5-HT, the effect of the mutant backgrounds on the response was less pronounced. Overall, the data support a role for cAMP in modulating the excitatory action of both FLP17A and 5-HT on C. elegans pharyngeal pumping and furthermore implicate an EGL-30 dependent pathway in the regulation of the response to FLP17A. [ABSTRACT FROM AUTHOR]

Abstract   Objective To explore the views and experiences of pharmacists and their mentoring designated medical practitioners (DMPs) about the ‘period of learning in practice’ (PLP) as part of supplementary prescribing (SP) training. Method Two focus groups (n = 5 and 7) of SP pharmacists were organised in Scotland. The experiences and views of DMPs (n = 13) were explored using one-to-one telephone interviews. The focus groups and interviews were transcribed verbatim and analysed using the framework approach. Main outcome measures Views and experiences of pharmacists and DMPs about the PLP. Results Planning the PLP in consultation with the DMP was found to be crucial for an optimal learning experience. Pharmacists who did not have a close working relationship with the medical team had difficulties in identifying a DMP and organising their PLP. Participants stressed the importance of focusing on and achieving the core competencies for prescribers during the PLP. Input from doctors involved in the training of others, review of consultation videos, and formal independent assessment including clinical assessment at the end of the PLP might improve the quality of the PLP. Forums for discussing experiences during the PLP and gathering information might be valuable. Conclusion Our findings have implications for prescribing training for pharmacists in the future. The PLP should focus on core competencies with input from doctors involved in the training of others and have a formal assessment of consultation skills. Support for pharmacists in organising the PLP and forums for discussing experiences during the PLP would be valuable. [ABSTRACT FROM AUTHOR]

Abstract   Objective The aims of the present study were to: analyse the prevalence of polypharmacy in a group of older patients; evaluate the influence of hospital stay on the number of drugs taken; assess the most frequently prescribed pharmacological classes; identify risk factors that predisposed the patient to polypharmacy. Setting The study was carried out in the Department of Internal Medicine of a non-university general hospital. Method In the retrospective study, 600 patients aged 65 years or more were enrolled. They were hospitalised in the period from 1st December 2003 to 31st March 2005. Each person taking six or more medications per day was considered to be a patient with polypharmacy. Particular sociodemographic and clinical characteristics, as well as comorbid conditions, were evaluated as factors potentially influencing the prevalence of polypharmacy. Main outcome measure The number and type of medications taken at the time of hospital admission and discharge were recorded and compared for each patient. Results Polypharmacy on admission and at discharge was observed in 362 (60.3%) and 374 (62.3%) patients, respectively. Hospitalisation led to a significant increase in the number of medications. The spectrum of medications used corresponded to the proportions of diagnoses in the evaluated group, in which cardiovascular diseases were most prevalent. According to the multivariate analysis using a logistic regression model, diabetes mellitus (odds ratio (OR) 2.40; 95% confidence interval (CI): 1.64–3.50), heart failure (OR 2.14; 95% CI: 1.46–3.14), dementia (OR 2.12; 95% CI: 1.26–3.57), living alone (OR 2.00; 95% CI: 1.28–3.10), arterial hypertension (OR 1.63; 95% CI: 1.08–2.44) and cerebrovascular disease (OR 1.58; 95% CI: 1.03–2.44) significantly increased the risk of the presence of polypharmacy. Conclusion Our study confirmed a relatively high prevalence of polypharmacy in Slovak elderly patients. Polypharmacy risk rose especially with the increased prevalence of diseases of advancing age (diabetes mellitus, heart failure, arterial hypertension, dementia and cerebrovascular diseases). The increasing numbers of medications in inpatients indicate the need for the careful re-evaluation of pharmacotherapy during the stay in hospital. [ABSTRACT FROM AUTHOR]

Serotonin (5-HT) regulates a wide range of behaviors in Caenorhabditis elegans, including egg laying, male mating, locomotion and pharyngeal pumping. So far, four serotonin receptors have been described in the nematode C. elegans, three of which are G protein-coupled receptors (GPCR), (SER-1, SER-4 and SER-7), and one is an ion channel (MOD-1). By searching the C. elegans genome for additional 5-HT GPCR genes, we identified five further genes which encode putative 5-HT receptors, based on sequence similarities to 5-HT receptors from other species. Using loss-of-function mutants and RNAi, we performed a systematic study of the role of the eight GPCR genes in serotonin-modulated behaviors of C. elegans (F59C12.2, Y22D7AR.13, K02F2.6, C09B7.1, M03F4.3, F16D3.7, T02E9.3, C24A8.1). We also examined their expression patterns. Finally, we tested whether the most likely candidate receptors were able to modulate adenylate cyclase activity in transfected cells in a 5-HT-dependent manner. This paper is the first comprehensive study of G protein-coupled serotonin receptors of C. elegans. It provides a direct comparison of the expression patterns and functional roles for 5-HT receptors in C. elegans. [ABSTRACT FROM AUTHOR]

Abstract  Invertebrate neuroscience has provided a number of very informative model systems that have been extensively utilized in order to define the neurobiological bases of animal behaviours (Sattelle and Buckingham in Invert Neurosci 6:1–3, 2006). Most eminent among these are a number of molluscs, including Aplysia californica, Lymnaea stagnalis and Helix aspersa, crustacean systems such as the crab stomatogastric ganglion and a wide-range of other arthropods. All of these have been elegantly exploited to shed light on the very important phenomenon of the molecular and cellular basis for synaptic regulation that underpins behavioural plasticity. Key to the successful use of these systems has been the ability to study well-defined, relatively simple neuronal circuits that direct and regulate a quantifiable animal behaviour. Here we describe the pharyngeal system of the nematode C. elegans and its utility as a model for defining the genetic basis of behaviour. The circuitry of the nervous system in this animal is uniquely well-defined. Furthermore, the feeding behaviour of the worm is controlled by the activity of the pharynx and this in turn is regulated in a context-dependent manner by a simple nervous system that integrates external signals, e.g. presence or absence of food, and internal signals, e.g. the nutritional status of the animal to direct an appropriate response. The genetics of C. elegans is being effectively exploited to provide novel insight into genes that function to regulate the neuronal network that controls the pharynx. Here we summarise the progress to date and highlight topics for future research. Two main themes emerge. First, although the anatomy of the pharyngeal system is very well-defined, there is a much poorer understanding of its neurochemistry. Second, it is evident that the neurochemistry is remarkably complex for such a simple circuit/behaviour. This suggests that the pharyngeal activity may be subject to exquisitely precise regulation depending on the animal’s environment and status. This therefore provides a very tractable genetic model to investigate neural mechanisms for signal integration and synaptic plasticity in a well-defined neuronal network that directs a quantifiable behaviour, feeding. [ABSTRACT FROM AUTHOR]

Abstract

The metabotropic glutamate receptors (mGluRs) are a class of G-protein-coupled receptor that undergo extensive interactions with scaffolding proteins, and this is intrinsic to their function as an important group of neuromodulators at glutamatergic synapses. The Caenorhabditis elegans nervous system expresses three metabotropic glutamate receptors, MGL-1, MGL-2 and MGL-3. Relatively little is known about how the function and signalling of these receptors is organised in C. elegans. To identify proteins that scaffold the MGL-1 receptor, we have conducted a yeast two-hybrid screen. Three of the interacting proteins, MPZ-1, NRFL-1 and PTP-1, displayed motifs characteristic of mammalian mGluR scaffolding proteins. Using cellular co-expression criterion, we show mpz-1 and ptp-1 exhibited overlapping expression patterns with subsets of mgl-1 neurons. This included neurones in the pharyngeal nervous system that control the feeding organ of the worm. The mGluR agonist L-CCG-I inhibits the activity of this network in wild-type worms, in an MGL-1 and dose-dependent manner. We utilised L-CCG-I to identify if MGL-1 function was disrupted in mutants with deletions in the mpz-1 gene. The mpz-1 mutants displayed a largely wild-type response to L-CCG-I, suggesting MGL-1 signalling is not overtly disrupted consistent with a non-obligatory modulatory function in receptor scaffolding. The selectivity of the protein interactions and overlapping expression identified here warrant further investigation of the functional significance of scaffolding of metabotropic glutamate receptor function. [ABSTRACT FROM AUTHOR]