Regulation of photosynthetic electron flow on dark to light transition by ferredoxin:NADP(H) oxidoreductase interactions.

During photosynthesis, electron transport is necessary for carbon assimilation and must be regulated to minimize free radical damage. There is a longstanding controversy over the role of a critical enzyme in this process (ferredoxin:NADP(H) oxidoreductase, or FNR), and in particular its location within chloroplasts. Here we use immunogold labelling to prove that FNR previously assigned as soluble is in fact membrane associated. We combined this technique with a genetic approach in the model plant Arabidopsis to show that the distribution of this enzyme between different membrane regions depends on its interaction with specific tether proteins. We further demonstrate a correlation between the interaction of FNR with different proteins and the activity of alternative photosynthetic electron transport pathways. This supports a role for FNR location in regulating photosynthetic electron flow during the transition from dark to light.

The prevalence of constant supportive observations in high, medium and low secure services.

Aims and method We explored the prevalence and use of constant supportive observations (CSO) in high, medium and low secure in-patient services in a single National Health Service (NHS) mental health trust. From clinical records, we extracted data on the length of time of CSO, the reason for the initiation of CSO and associated adverse incidents for all individuals who were placed on CSO between July 2013 and June 2014. RESULTS: A small number of individuals accounted for a disproportionately large proportion of CSO hours in each setting. Adverse incident rates were higher on CSO than when not on CSO. There was considerable variation between different settings in terms of CSO use and the reasons for commencing CSO. Clinical implications The study describes the prevalence and nature of CSO in secure forensic mental health services and the associated organisational costs. The marked variation in CSO use between settings suggests that mental health services continue to face challenges in balancing risk management with minimising restrictive interventions. Declaration of interest A.B. and J.L.I. are both directly employed by the NHS trust in which the study was conducted.

An evaluation of subjective experiences, effects and overall satisfaction with clozapine treatment in a UK forensic service.

OBJECTIVES: Patients prescribed clozapine were surveyed to assess (a) the effects, both positive and adverse, and overall satisfaction with clozapine in comparison to previously prescribed antipsychotics and (b) the relative significance of effects experienced, both positive and adverse, in terms of impact on subjective well-being. METHODS: A total of 56 male patients prescribed clozapine at a forensic psychiatric hospital were surveyed using a 27-item questionnaire. All patients had been prescribed clozapine for a minimum of 3 months. Respondents were asked to rate effects and satisfaction with clozapine treatment in comparison with previously prescribed antipsychotic medication on a five-point scale. Respondents were also asked to rate effects experienced with clozapine treatment in terms of impact on subjective well-being on a five-point scale. RESULTS: A total of 89% of respondents reported greater satisfaction with clozapine than with previously prescribed antipsychotic medication. A majority of patients reported positive effects in terms of an improvement in their quality of life (68%) and social abilities (52%) with clozapine in comparison with previously prescribed antipsychotics. Nocturnal hypersalivation (84%) and weight gain (57%) were the most common adverse effects. Hedonic responses were assessed for each effect in order to determine the associated subjective experiences. The most positive hedonic responses were for quality of life, mood and alertness. In terms of adverse impact on subjective well-being, nocturnal hypersalivation ranked highest. CONCLUSIONS: Patients in a UK forensic sample are largely satisfied with clozapine treatment. The subjective effects of clozapine treatment should be taken into account by clinicians when assessing response. This may provide an opportunity to highlight the positive changes and prioritize management of the most undesirable adverse effects, which is likely to promote compliance and improve longer term treatment outcomes.

Evaluation of the clinical maxim: "If it ain't broke, don't fix it".

PURPOSE: A significant number of patients return to optometric practice dissatisfied with their spectacles. An important question is whether any of these cases are preventable. There are several different clinical maxims that are used to modify the subjective refraction when determining the refractive prescription. These maxims aim to improve patient comfort and adaptation and thereby reduce patient dissatisfaction with new spectacles. They are not based on research evidence, but rather on expert opinion gained from clinical experience. The aim of this study was to retrospectively analyze a large number of case records of dissatisfied patients to assess the possible usefulness of the prescribing maxim "if it ain't broke, don't fix it." METHODS: Three hundred eighteen non-tolerance cases from a university-based Canadian optometric clinic were categorized by a focus group of optometrists. Three prescribing categories were defined and comprised cases in which application of the proposed maxim may have prevented the recheck eye examination; a more limited application of the maxim for one working distance may have been appropriate; and finally scenarios in which the maxim did not work in that the practitioner was judged to have initially followed the maxim, yet patient dissatisfaction was still reported. The remaining unallocated records comprised prescribing situations outside the scope of this study. RESULTS: Approximately 32% of non-tolerance cases were judged to have been preventable by use of the proposed maxim. Furthermore, an additional 10% reduction in recheck cases may have been possible by a more liberal interpretation of the maxim. Conversely, 4% of cases were deemed to comprise scenarios in which the maxim was followed yet the patient returned later to report problems with their spectacles. CONCLUSIONS: The prescribing maxim "if it ain't broke, don't fix it" appears to have a role in reducing recheck eye examinations and improving patient satisfaction with new spectacles.

Spectacle prescribing II: practitioner experience is linked to the likelihood of suggesting a partial prescription.

PURPOSE: A follow up study to investigate whether UK optometrists partially prescribe significant changes in refractive correction to assist patient adaptation and whether various aspects of practitioner profiles are linked to the nature of these prescribing decisions. METHOD: A case scenario type questionnaire was distributed by post and via the internet to UK optometrists. Five case scenarios were described that included information on patient age, symptoms, habitual refractive correction (if any), subjective refraction and any other relevant clinical information. In each case respondents were asked to indicate and justify what refractive correction they would prescribe. RESULTS: A total of 592 questionnaires were completed. Between 41% and 84% prescribed the subjective refraction result depending on the scenario. The likelihood of partial prescribing increased by 34% for every 10 years following qualification and thus after a typical 40 year career, respondents were now over three times more likely to partially prescribe. There were no other links with the propensity to partially prescribe. CONCLUSION: The subjective refraction result exerted a strong hold on the reported prescribing outcome, particularly for newly qualified optometrists. Partial prescribing was increasingly proposed the greater the number of years the respondent had been qualified. This suggests that with increasing exposure to patients who return dissatisfied with their spectacles, a greater appreciation of partial prescribing is gained. This link seems to be an important finding that provides significant support for the prescribing rules suggested by textbooks, which are not yet supported by research evidence.

What adjustments, if any, do UK optometrists make to the subjective refraction result prior to prescribing?

AIM: To determine whether UK optometrists routinely prescribe the subjective refraction result, or whether adjustments are made to aid patient comfort and adaptation. METHOD: A questionnaire was distributed by post and via the internet to UK optometrists. It incorporated eight case scenarios that included information regarding a patient's age, symptoms, habitual refractive correction, subjective refraction and any other pertinent clinical information. For each case, they were asked to indicate what refractive correction they would prescribe. RESULTS: A total of 426 questionnaires were completed. In three cases with asymptomatic patients with negligible changes in visual acuity, about 45% of respondents indicated that they would prescribe the subjective refraction result and about 40% would prescribe the habitual correction. In a further case involving a 75 year-old asymptomatic patient, 55% prescribed the habitual correction. In two cases with large cylinder power changes, about 40% of respondents indicated they would partially prescribe the cylinder power. However, there was a large variation in the suggested modification of sphere powers to accompany the partial cylinder changes. CONCLUSION: A large sample of UK optometrists indicated that they would prescribe the subjective refraction result regardless of patient age, symptoms and difference between habitual correction and subjective refraction. Optometrists who partially prescribe, employ a wide variety of techniques including some that conflict with conventional teaching.

Relative analgesic potencies of levobupivacaine and ropivacaine for caudal anesthesia in children.

BACKGROUND: Comparing relative potency of new local anesthetics, such as levobupivacaine and ropivacaine, by the minimum local analgesic concentration model has not been described for caudal anesthesia. Therefore, we performed a prospective, randomized, double-blind study to determine the minimum local analgesic concentrations of a caudal single shot of ropivacaine and levobupivacaine in children and to describe the upper dose-response curve. METHODS: We performed a two-stage prospective, randomized, double-blind study comparing the dose-response curves of caudal ropivacaine and levobupivacaine in children. In phase 1, 80 boys were randomized to receive either ropivacaine or levobupivacaine. In the second phase a further 32 patients were randomly allocated to receive caudal anesthesia with doses designed to delineate the upper dose-response range (the 50% effective dose [ED(50)]-ED(95) range). RESULTS: There were no significant differences in ED(50) values for caudal ropivacaine and levobupivacaine. The ED(50) for levobupivacaine estimated from the Dixon Massey method was 0.069% (95% CI 0.056%-0.082%) and for ropivacaine was 0.075% (95% CI 0.058%-0.092%). Estimated by isotonic regression the ED(50) and ED(95) respectively of levobupivacaine were 0.068 (0.04-0.09) and 0.20% (95% CI 0.16%-0.24%). For ropivacaine ED 50 and ED95 were 0.066 (0.033-0.098) and 0.225% (95% CI 0.21%-0.24%). CONCLUSIONS: In children receiving one minimum alveolar anesthetic concentration of sevoflurane, there were no significant differences in the ED(50) for caudal levobupivacaine and ropivacaine. The potency ratio at ED(50) was 0.92 and 0.89 at ED(95), indicating that caudal levobupivacaine and ropivacaine have a similar potency.

Biodistribution and toxicological safety of adenovirus type 5 and type 35 vectored vaccines against human immunodeficiency virus-1 (HIV-1), Ebola, or Marburg are similar despite differing adenovirus serotype vector, manufacturer's construct, or gene inserts.

The Vaccine Research Center has developed vaccine candidates for different diseases/infectious agents (including HIV-1, Ebola, and Marburg viruses) built on an adenovirus vector platform, based on adenovirus type 5 or 35. To support clinical development of each vaccine candidate, pre-clinical studies were performed in rabbits to determine where in the body they biodistribute and how rapidly they clear, and to screen for potential toxicities (intrinsic and immunotoxicities). The vaccines biodistribute only to spleen, liver (Ad5 only), and/or iliac lymph node (Ad35 only) and otherwise remain in the site of injection muscle and overlying subcutis. Though approximately 10(11) viral particles were inoculated, already by Day 9, all but 10(3) to 10(5) genome copies per mu g of DNA had cleared from the injection site muscle. By three months, the adenovector was cleared with, at most, a few animals retaining a small number of copies in the injection site, spleen (Ad5), or iliac lymph node (Ad35). This pattern of limited biodistribution and extensive clearance is consistent regardless of differences in adenovector type (Ad5 or 35), manufacturer's construct and production methods, or gene-insert. Repeated dose toxicology studies identified treatment-related toxicities confined primarily to the sites of injection, in certain clinical pathology parameters, and in body temperatures (Ad5 vectors) and food consumption immediately post-inoculation. Systemic reactogenicity and reactogenicity at the sites of injection demonstrated reversibility. These data demonstrate the safety and suitability for investigational human use of Ad5 or Ad35 adenovector-based vaccine candidates at doses of up to 2 x 10(11) given intramuscularly to prevent various infectious diseases.

Gene expression analysis of troglitazone reveals its impact on multiple pathways in cell culture: a case for in vitro platforms combined with gene expression analysis for early (idiosyncratic) toxicity screening.

Peroxisome proliferator-activated receptor gamma (PPARgamma) agonists of the thiazolidinedione family are used for the treatment of type 2 diabetes mellitus due to their ability to reduce glucose and lipid levels in patients with this disease. Three thiazolidinediones that were approved for treatment are Rezulin (troglitazone), Avandia (rosiglitazone), and Actos (pioglitazone). Troglitazone was withdrawn from the market due to idiosyncratic drug toxicity. Rosiglitazone and pioglitazone are still on the market for the treatment of type 2 diabetes. The authors present data from a gene expression screen that compares the impact these three compounds have in rats, in rat hepatocytes, and in the clone 9 rat liver cell line. The authors monitored the changes in expression in multiple genes, including those related to xenobiotic metabolism, proliferation, DNA damage, oxidative stress, apoptosis, and inflammation. Compared to the other two compounds, troglitazone had a significant impact on many of the pathways monitored in vitro although no major perturbation was detected in vivo. The changes detected predict not only general toxicity but potential mechanisms of toxicity. Based on gene expression analysis, the authors propose there is not just one but multiple ways troglitazone could be toxic, depending on a patient's environment and genetic makeup, including immune response-related toxicity.

Biodistribution of DNA plasmid vaccines against HIV-1, Ebola, Severe Acute Respiratory Syndrome, or West Nile virus is similar, without integration, despite differing plasmid backbones or gene inserts.

The Vaccine Research Center has developed a number of vaccine candidates for different diseases/infectious agents (HIV-1, Severe Acute Respiratory Syndrome virus, West Nile virus, and Ebola virus, plus a plasmid cytokine adjuvant-IL-2/Ig) based on a DNA plasmid vaccine platform. To support the clinical development of each of these vaccine candidates, preclinical studies have been performed in mice or rabbits to determine where in the body these plasmid vaccines would biodistribute and how rapidly they would clear. In the course of these studies, it has been observed that regardless of the gene insert (expressing the vaccine immunogen or cytokine adjuvant) and regardless of the promoter used to drive expression of the gene insert in the plasmid backbone, the plasmid vaccines do not biodistribute widely and remain essentially in the site of injection, in the muscle and overlying subcutis. Even though approximately 10(14) molecules are inoculated in the studies in rabbits, by day 8 or 9 ( approximately 1 week postinoculation), already all but on the order of 10(4)-10(6) molecules per microgram of DNA extracted from tissue have been cleared at the injection site. Over the course of 2 months, the plasmid clears from the site of injection with only a small percentage of animals (generally 10-20%) retaining a small number of copies (generally around 100 copies) in the muscle at the injection site. This pattern of biodistribution (confined to the injection site) and clearance (within 2 months) is consistent regardless of differences in the promoter in the plasmid backbone or differences in the gene insert being expressed by the plasmid vaccine. In addition, integration has not been observed with plasmid vaccine candidates inoculated i.m. by Biojector 2000 or by needle and syringe. These data build on the repeated-dose toxicology studies performed (see companion article, Sheets et al., 2006) to demonstrate the safety and suitability for investigational human use of DNA plasmid vaccine candidates for a variety of infectious disease prevention indications.