Graphene-oxide loading on natural zeolite particles for enhancement of adsorption properties.

Multiple methods of grafting graphene oxide (GO) nanosheets to natural clinoptilolite-rich zeolite particles were developed in our laboratory. In this study, we have systematically characterized the GO coated particles prepared by various methods to select the most promising method for further research efforts. This study revealed that the most promising coating method was the clean-acid-treated zeolite particles followed by deposition of GO nanosheets onto the zeolite surface and mild thermal treatment of the particles. GO and its synergistic interaction in zeolite was attributed to electrostatic interactions, hydrophobic interactions and hydrogen bonds. Hydrophobic interactions are enhanced both due to dealumination of zeolite caused by the cleaning method followed by acid treatment and due to partial thermal deoxygenation of GO. This method provided a ten times larger surface area (from 10.55 m2 g-1 to 117.96 m2 g-1) and three times smaller pore diameter (from 81.91 Å to 30.68 Å), providing great particles for a variety of applications as adsorbents or catalysts.

Mucosal-associated invariant T cells and Vδ2+ γδ T cells in community acquired pneumonia: association of abundance in sputum with clinical severity and outcome.

Mucosal-associated invariant T (MAIT) cells and Vδ2+ γδ T cells are anti-bacterial innate-like lymphocytes (ILLs) that are enriched in blood and mucosa. ILLs have been implicated in control of infection. However, the role of ILLs in community-acquired pneumonia (CAP) is unknown. Using sputum samples from a well-characterized CAP cohort, MAIT cell and Vδ2+ T cell abundance was determined by quantitative polymerase chain reaction (qPCR). Cytokine and chemokine concentrations in sputum were measured. The capacity of bacteria in sputum to produce activating ligands for MAIT cells and Vδ2+ T cells was inferred by 16S rRNA sequencing. MAIT cell abundance in sputum was higher in patients with less severe pneumonia; duration of hospital admission was inversely correlated with both MAIT and Vδ2+ T cell abundance. The abundance of both ILLs was higher in patients with a confirmed bacterial aetiology; however, there was no correlation with total bacterial load or the predicted capacity of bacteria to produce activating ligands. Sputum MAIT cell abundance was associated with interferon (IFN)-α, IFN-γ, and sputum neutrophil abundance, while Vδ2+ T cell abundance was associated with CXCL11 and IFN-γ. Therefore, MAIT and Vδ2+ T cells can be detected in sputum in CAP, where they may contribute to improved clinical outcome.

Generation of transgenic goats by pronuclear microinjection: a retrospective analysis of a commercial operation (1995-2012).

Production of transgenic founder goats involves introducing and stably integrating an engineered piece of DNA into the genome of the animal. At LFB USA, the ultimate use of these transgenic goats is for the production of recombinant human protein therapeutics in the milk of these dairy animals. The transgene or construct typically links a milk protein specific promoter sequence, the coding sequence for the gene of interest, and the necessary downstream regulatory sequences thereby directing expression of the recombinant protein in the milk during the lactation period. Over the time period indicated (1995-2012), pronuclear microinjection was used in a number of programs to insert transgenes into 18,120, 1- or 2- cell stage fertilized embryos. These embryos were transferred into 4180 synchronized recipient females with 1934 (47%) recipients becoming pregnant, 2594 offspring generated, and a 109 (4.2%) of those offspring determined to be transgenic. Even with new and improving genome editing tools now available, pronuclear microinjection is still the predominant and proven technology used in this commercial setting supporting regulatory filings and market authorizations when producing founder transgenic animals with large transgenes (> 10 kb) such as those necessary for directing monoclonal antibody production in milk.

Motivational and behavioural models of change: A longitudinal analysis of change among men with chronic haemophilia-related joint pain.

BACKGROUND: Motivational and behavioural models of adjustment to chronic pain make different predictions about change processes, which can be tested in longitudinal analyses. METHODS: We examined changes in motivation, coping and acceptance among 78 men with chronic haemophilia-related joint pain. Using cross-lagged regression analyses of changes from baseline to 6 months as predictors of changes from 6 to 12 months, with supplementary structural equation modelling, we tested two models in which motivational changes influence behavioural changes, and one in which behavioural changes influence motivational changes. RESULTS: Changes in motivation to self-manage pain influenced later changes in pain coping, consistent with the motivational model of pain self-management, and also influenced later changes in activity engagement, the behavioural component of pain acceptance. Changes in activity engagement influenced later changes in pain willingness, consistent with the behavioural model of pain acceptance. CONCLUSIONS: Based on the findings, a combined model of changes in pain self-management and acceptance is proposed, which could guide combined interventions based on theories of motivation, coping and acceptance in chronic pain. SIGNIFICANCE: This study adds longitudinal evidence about sequential change processes; a test of the motivational model of pain self-management; and tests of behavioural versus motivational models of pain acceptance.

A double-blind, randomized, placebo-controlled, parallel group study of THC/CBD spray in peripheral neuropathic pain treatment.

BACKGROUND: Peripheral neuropathic pain (PNP) associated with allodynia poses a significant clinical challenge. The efficacy of Δ(9) -tetrahydrocannabinol/cannabidiol (THC/CBD) oromucosal spray, a novel cannabinoid formulation, was investigated in this 15-week randomized, double-blind, placebo-controlled parallel group study. METHODS: In total, 303 patients with PNP associated with allodynia were screened; 128 were randomized to THC/CBD spray and 118 to placebo, in addition to their current analgesic therapy. The co-primary efficacy endpoints were the 30% responder rate in PNP 0-10 numerical rating scale (NRS) score and the mean change from baseline to the end of treatment in this score. Various key secondary measures of pain and functioning were also investigated. RESULTS: At the 30% responder level, there were statistically significant treatment differences in favour of THC/CBD spray in the full analysis (intention-to-treat) dataset [p = 0.034; 95% confidence interval (CI): 1.05-3.70]. There was also a reduction in mean PNP 0-10 NRS scores in both treatment groups that was numerically higher in the THC/CBD spray group, but which failed to reach statistical significance. Secondary measures of sleep quality 0-10 NRS score (p = 0.0072) and Subject Global Impression of Change (SGIC) (p = 0.023) also demonstrated statistically significant treatment differences in favour of THC/CBD spray treatment. CONCLUSIONS: These findings demonstrate that, in a meaningful proportion of otherwise treatment-resistant patients, clinically important improvements in pain, sleep quality and SGIC of the severity of their condition are obtained with THC/CBD spray. THC/CBD spray was well tolerated and no new safety concerns were identified.

Atypical guanylyl cyclase from the pond snail Lymnaea stagnalis: cloning, sequence analysis and characterization of expression.

Soluble guanylyl cyclases (sGCs) are traditionally recognized as the main molecular receptor for nitric oxide (NO), a gaseous transmitter involved in many functions of the nervous system. Some sGCs are however insensitive to NO and therefore are known as atypical. Although atypical sGCs have been shown to exist in both vertebrate and invertebrate nervous systems, our understanding of their functional role is incomplete. Here we report on the cloning, sequencing and localization of an atypical sGC named Lym-sGCbeta3 from the snail Lymnaea stagnalis. We found that Lym-sGCbeta3 shares a number of structural characteristics with some previously characterized atypical sGCs including the presence of Tyr140 in the regulatory domain. This residue is thought to be of a critical importance in determining sensitivity of atypical sGCs to oxygen. These findings raise the possibility that Lym-sGCbeta3 is an oxygen receptor. The results of our in situ hybridization and RT-PCR experiments support this idea further by showing that Lym-sGCbeta3 is expressed in the osphradium, a peripheral sense organ in which oxygen-sensing neurons are located. Also of interest are our observations that many neurons in Lymnaea CNS co-express conventional and atypical sGC subunits. These data are consistent with a possible dominant negative regulatory role of atypical sGC subunits through the formation of heterodimers exhibiting low enzymatic activity.

Direct imaging of nanoscale phase separation in La(0.55)Ca(0.45)MnO(3): relationship to colossal magnetoresistance.

A nanoscale phase is known to coincide with colossal magnetoresistance (CMR) in manganites, but its volume fraction is believed to be too small to affect CMR. Here we provide scanning-electron-nanodiffraction images of nanoclusters as they form and evolve with temperature in La(1-x)Ca(x)MnO(3), x = 0.45. They are not doping inhomogeneities, and their structure is that of the bulk compound at x = 0.60, which at low temperatures is insulating. Their volume fraction peaks at the CMR critical temperature and is estimated to be 22% at finite magnetic fields. In view of the known dependence of the nanoscale phase on magnetic fields, such a volume fraction can make a significant contribution to the CMR peak.

Tools to share good chairside teaching practice: a clinical scenario and appreciative questionnaire.

This article provides a scenario for analysis of good chairside teaching practice to serve as a starting point for continued discussion in this complex field. Documented issues of good chairside teaching practice are cross-referenced to a clinical scenario with explanations in the form of a commentary. This provided the context for generating a set of questions that are provided as tools to support good chairside practice. These tools are designed to be used with 'Appreciative Inquiry', which claims that there is much to be gained by discovering where excellence is possible and elaborating upon this. Although this process can be carried out in single units or departments, it is proposed that collaboration between institutions would allow sharing of valuable innovations and greater understanding of educational training, production of good practice guidance and professional development of staff. This article is the third in a series of three and provides a scaffold for a scenario and questions to encourage collaboration in evolving and sharing good chairside teaching practice. The first article investigated the perceptions of stakeholders in chairside teaching at a single dental school and the second evaluated chairside teaching on a UK wide scale. A further accompanying article reviews some of the educational methodology and innovations in teaching and learning that may be applied to dentistry.

Characterization of JEOL 2100F Lorentz-TEM for low-magnification electron holography and magnetic imaging.

We present results that characterize the performance and capabilities of the JEOL 2100F-LM electron microscope to carry out holography and quantitative magnetic imaging. We find the microscope is well-suited for studies of magnetic materials, or for semi-conductor dopant profiling, where a large hologram width ( approximately 1 microm) and fine fringe spacing ( approximately 1.5 nm) are obtained with good contrast ( approximately 20%). We present, as well, measurements of the spherical aberration coefficient Cs=(108.7+/-9.6)mm and minimum achievable focal step delta f=(87.6+/-1.4)nm for the specially designed long-focal-length objective lens of this microscope. Further, we detail experiments to accurately measure the optical parameters of the imaging system typical of conventional holography setup in a transmission electron microscope. The role played by astigmatic illumination in the hologram formation is also assessed with a wave-optical model, which we present and discuss. The measurements obtained for our microscope are used to simulate realistic holograms, which we compare directly to experimental holograms finding good agreement. These results indicate the usefulness of measuring these optical parameters to guide the optimization of the experimental setup for a given microscope, and to provide an additional degree of practical experimental possibility.

Self-reported and clinically determined oral health status predictors for quality of life in dentate older migrant adults.

OBJECTIVE: This paper reports the impact of oral health on the quality of life (QOL) of Southern European, dentate older adults, living independently in Melbourne, Australia. Participants were recruited through ethnic social clubs and interviewed about oral health, general health, socio-demographics, and QOL using the Medical Outcomes Study Short-Form 12 (SF-12). The SF-12's physical and mental health component summary scores (PCS and MCS, respectively) were computed. The Oral Health Impact Profile (OHIP-14) assessed the specific impact of oral health on QOL. Participants were also given a clinical oral examination. RESULTS: A total of 603 eligible older adults volunteered; 308 were from Greek background and 295 were from Italian background. Mean age was 67.7 years (SD 6.2), with 63.7% being female. The PCS score had a mean value of 45.8 (SD 11.8), and MCS had a mean of 47.8 (SD 5.7). PCS was associated with, periodontal status, chronic health condition, self-perceived oral health needs, self-assessed oral health status, oral health impact score and the interaction between gender and level of education [F(11 552) = 10.57; P < 0.0001]. These independent variables accounted for 16% of the variance in PCS. The multivariate model predicting MCS had only one significant variable (self-reported gingival bleeding), explaining 1.5% of the variance. The OHIP-14 ranged from 0 to 48 with a mean score of 5.6 (SD 9.3). The model predicting OHIP-14 contained four significant variables: perceived oral health treatment needs, number of missing natural teeth, reports of having to sip liquid to help swallow food, and gender [F(4576) = 33.39; P < 0.0001], and explained 18% of the variance. The results demonstrated a negative association between oral health indicators and both the oral health-related QOL and the physical component of the SF-12. CONCLUSION: The present findings support a growing recognition of the importance of oral health as a mediator of QOL. However, the self-selected sample and modest predictive power of the multivariate models suggest that further research is needed to expand this explanatory model.

Polaron melting and ordering as key mechanisms for colossal resistance effects in manganites.

Polarons, the combined motion of electrons in a cloth of their lattice distortions, are a key transport feature in doped manganites. To develop a profound understanding of the colossal resistance effects induced by external fields, the study of polaron correlations and the resulting collective polaron behavior, i.e., polaron ordering and transition from polaronic transport to metallic transport is essential. We show that static long-range ordering of Jahn-Teller polarons forms a polaron solid which represents a new type of charge and orbital ordered state. The related noncentrosymmetric lattice distortions establish a connection between colossal resistance effects and multiferroic properties, i.e., the coexistence of ferroelectric and antiferromagnetic ordering. Colossal resistance effects due to an electrically induced polaron solid-liquid transition are directly observed in a transmission electron microscope with local electric stimulus applied in situ using a piezo-controlled tip. Our results shed light onto the colossal resistance effects in magnetic field and have a strong impact on the development of correlated electron-device applications such as resistive random access memory (RRAM).

A straightforward specimen holder modification for remnant magnetic-field measurement in TEM.

We report a simple means to modify an analytic sample holder to perform ambient Hall probe measurements of the sample area inside a transmission electron microscope (TEM). These measurements are important in the case of electron microscopy studies involving magnetic materials. We characterize the magnetic field of the JEOL 2100F-LM, a microscope dedicated in design to perform magnetic imaging, and also of the JEOL 3000F FEG-TEM operated in Lorentz mode. In the case of the 3000F, we measure vertical remnant field about 300 Oe due to the objective lens of the microscope. In the case of the 2100F, design of the objective lens reduces the remnant field to about 4 Oe. We characterize the field along two orthogonal directions, and spatial characteristics of the field profile are made for both microscopes during all stages of specimen entry into the column. In the case of the 2100F, we additionally measure the field conditions as a function of objective lens excitation, which is important for in situ magnetization experiments. Finally, we provide experimental results illustrating the importance of these measurements.

MRI findings following laparoscopic sacrocolpopexy.

Sacrocolpopexy is a surgical procedure that provides effective treatment for pelvic prolapse. The surgical technique and complications of laproscopic sacrocolpopexy are described. This review presents experience of MRI for postoperative assessment and illustrates normal and abnormal findings.

On the transport of intensity technique for phase retrieval.

The Transport of Intensity technique is becoming a viable alternative to electron holography for phase retrieval in Transmission Electron Microscopy. However, several issues are still to be clarified in order to ascertain the applicability of the technique; among them, the controversy regarding its geometrical or wave-optical nature, as related to the phase detection limit. We show here that the Transport of Intensity is a wave-optical technique that works in a special regime of small defocus where the image intensity is linear with the defocus parameter. By a simple analytical example we show that the Transport of Intensity correctly reconstructs the electron optical phase shift even when the phase is smaller than pi, a value defining the boundary between the geometrical and wave approaches. Another example is given, the reconstruction of a phase jump, accompanied with experimental support showing that phase retrieval by Electron Holography and Transport of Intensity techniques yields results in good agreement.

A unique determination of boundary condition in quantitative electron diffraction: Application to accurate measurements of mean inner potentials.

We combine off-axis electron holography and electron shadow imaging to accurately determine the specimen thickness and the incident electron beam direction over the illuminated area of a crystal. We, furthermore, quantify the variations in diffraction intensity with position over the same area. This unique solution to the experimental boundary condition problem enables us to make precise measurements of mean inner electrostatic potentials and structure factors that are sensitive to the bonding characteristics of materials. In this paper, we present the results of mean-inner potential determination from silicon and the newly discovered magnesiumdiboride superconductor.

How well do we investigate patients with suspected subarachnoid haemorrhage? The continuing need for cerebrospinal fluid investigations.

OBJECTIVE: To demonstrate the extent of compliance with established guidelines for the investigation of suspected subarachnoid haemorrhage (SAH) and the implications of non-compliance. DESIGN: Prospective observational study of practice in three hospitals in the Trent region. SETTING: One teaching hospital with a tertiary neuroscience referral centre and two large district general hospitals. PARTICIPANTS: 50 consecutive patients from each centre referred for suspected SAH with negative computed tomography. MAIN OUTCOME MEASURES: Diagnosis of SAH confirmed or excluded according to guidelines. RESULTS: When the data from the three centres were combined (n = 150 computed tomography negative cases) cerebrospinal fluid (CSF) investigation was not performed in 60/150 (40%). In the 90 cases where CSF studies were performed SAH was confirmed in 11 (12%). CONCLUSION: There is significant non-compliance in following the established guidelines for the investigation of SAH at the centres studied. As the primary cause of non-traumatic SAH is ruptured aneurysm, which is associated with high morbidity and mortality from second haemorrhage, this highlights a major source of concern for clinical governance.

Evaluation of a Health Promoting Schools program to reduce smoking in Australian secondary schools.

This randomized controlled trial evaluated the effectiveness of a multicomponent Health Promoting Schools (HPS) intervention program in improving self-reported smoking outcomes among a cohort of adolescents in 22 public secondary schools in the Hunter Region of New South Wales, Australia. Pre-test surveys were completed by students in the first 2 years of secondary school, with a 2-year post-test survey. Multivariate analyses examined intervention effect for the main outcome, post-test smoking behavior, controlling for pre-test smoking status, school and other confounders. The sample comprised the cohort of 1852 students who completed both surveys. The results demonstrated that the HPS program failed to improve smoking behavior over the 2 years (equal increase of 10% in both groups). The program was successful in improving smoking knowledge, but not attitudes, in intervention versus control group (P < 0.001). Independent predictors of post-test smoking included: pre-test smoking [odds ratio (OR) = 5.44; 95% confidence interval (CI) = 3.20-9.28], being female (OR = 0.55; CI = 0.35-0.87), having more close friends who smoked (OR = 1.42; CI = 1.33-1.52), peer group having no clear opinion about smoking (OR = 3.23; CI = 1.27-8.27), having more positive and less negative attitudes towards smoking, and being less involved in school activities. We discuss methodological issues in multicomponent community-based interventions, and highlight the strengths and limitations of this study.

The Phe-X-Glu DNA binding motif of MutS. The role of hydrogen bonding in mismatch recognition.

The crystal structures of MutS protein from Thermus aquaticus and Escherichia coli in a complex with a mismatch-containing DNA duplex reveal that the Glu residue in a conserved Phe-X-Glu motif participates in a hydrogen-bonded contact with either an unpaired thymidine or the thymidine of a G-T base-base mismatch. Here, the role of hydrogen bonding in mismatch recognition by MutS is assessed. The relative affinities of MutS for DNA duplexes containing nonpolar shape mimics of A and T, 4-methylbenzimidazole (Z), and difluorotoluene (F), respectively, that lack hydrogen bonding donors and acceptors, are determined in gel mobility shift assays. The results provide support for an induced fit mode of mismatch binding in which duplexes destabilized by mismatches are preferred substrates for kinking by MutS. Hydrogen bonding between the O epsilon 2 group of Glu and the mismatched base contributes only marginally to mismatch recognition and is significantly less important than the aromatic ring stack with the conserved Phe residue. A MutS protein in which Ala is substituted for Glu(38) is shown to be defective for mismatch repair in vivo. DNA binding studies reveal a novel role for the conserved Glu residue in the establishment of mismatch discrimination by MutS.

Neuronal expression of an FMRFamide-gated Na+ channel and its modulation by acid pH.

The molluscan Phe-Met-Arg-Phe-amide (FMRFamide)-gated sodium channels (FaNaCs) show both structural and functional similarities to the mammalian acid-sensing ion channels (ASICs). Both channel types are related to the epithelial sodium channels and, although the neuropeptide FMRFamide directly gates the FaNaCs, it also modulates the proton-gating properties of ASICs. It is not yet known whether protons can alter the gating properties of the FaNaCs. We chose to examine this possibility at a site of FaNaC expression in the nervous system of the mollusk Lymnaea stagnalis. We cloned a putative L. stagnalis FaNaC (LsFaNaC) that exhibited a high degree of sequence identity to the Helix aspersa FaNaC (HaFaNaC, 60%), and a weaker homology to the ASICs (ASIC3, 22%). In situ hybridization was used to map the LsFaNaC expression pattern in the brain and to identify the right pedal giant1 (RPeD1) neuron as a site where the properties of the endogenous channel could be studied. In RPeD1 neurons isolated in culture, we demonstrated the presence of an FMRFamide-gated sodium current with features expected for a FaNaC: amiloride sensitivity, sodium selectivity, specificity for FMRFamide and Phe-Leu-Arg-Phe-amide (FLRFamide), and no dependency on G-protein coupling. The sodium current also exhibited rapid desensitization in response to repeated FMRFamide applications. Lowering of the pH of the bathing solution reduced the amplitude of the FMRFamide-gated inward current, while also activating an additional sustained weak inward current that was apparently not mediated by the FaNaC. Acidification also prevented the desensitization of the FMRFamide-induced inward current. The acid sensitivity of LsFaNaC is consistent with the hypothesis that FaNaCs share a common ancestry with the ASICs.

Interaction of Escherichia coli MutS and MutL at a DNA mismatch.

MutS and MutL are both required to activate downstream events in DNA mismatch repair. We examined the rate of dissociation of MutS from a mismatch using linear heteroduplex DNAs or heteroduplexes blocked at one or both ends by four-way DNA junctions in the presence and absence of MutL. In the presence of ATP, dissociation of MutS from linear heteroduplexes or heteroduplexes blocked at only one end occurs within 15 s. When both duplex ends are blocked, MutS remains associated with the DNA in complexes with half-lives of 30 min. DNase I footprinting of MutS complexes is consistent with migration of MutS throughout the DNA duplex region. When MutL is present, it associates with MutS and prevents ATP-dependent migration away from the mismatch in a manner that is dependent on the length of the heteroduplex. The rate and extent of mismatch-provoked cleavage at hemimethylated GATC sites by MutH in the presence of MutS, MutL, and ATP are the same whether the mismatch and GATC sites are in cis or in trans. These results suggest that a MutS-MutL complex in the vicinity of a mismatch is involved in activating MutH.