Human clinically non-functioning pituitary adenomas (NFAs) account for approximately 40% of diagnosed pituitary tumors. Epigenetic mutations in tumor suppressive genes play an important role in NFA development. Maternally expressed gene 3 (MEG3) is a long non-coding RNA (lncRNA) and we hypothesized that it is a candidate tumor suppressor whose epigenetic silencing is specifically linked to NFA development. In this study, we introduced MEG3 expression into PDFS cells, derived from a human NFA, using both inducible and constitutively active expression systems. MEG3 expression significantly suppressed xenograft tumor growth in vivo in nude mice. When induced in culture, MEG3 caused cell cycle arrest at the G1 phase. In addition, inactivation of p53 completely abolished tumor suppression by MEG3, indicating that MEG3 tumor suppression is mediated by p53. In conclusion, our data support the hypothesis that MEG3 is a lncRNA tumor suppressor in the pituitary and its inactivation contributes to NFA development.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.

For the past 10 years, animal health experts and human health experts have been gaining experience in the technical aspects of avian influenza in mostly separate fora. More recently, in 2006, in a meeting of the small WHO Working Group on Influenza Research at the Human Animal Interface (Meeting report available from: http://www.who.int/csr/resources/publications/influenza/WHO_CDS_EPR_GIP_2006_3/en/index.html) in Geneva allowed influenza experts from the animal and public health sectors to discuss together the most recent avian influenza research. Ad hoc bilateral discussions on specific technical issues as well as formal meetings such as the Technical Meeting on HPAI and Human H5N1 Infection (Rome, June, 2007; information available from: http://www.fao.org/avianflu/en/conferences/june2007/index.html) have increasingly brought the sectors together and broadened the understanding of the topics of concern to each sector. The sectors have also recently come together at the broad global level, and have developed a joint strategy document for working together on zoonotic diseases (Joint strategy available from: ftp://ftp.fao.org/docrep/fao/011/ajl37e/ajl37e00.pdf). The 2008 FAO-OIE-WHO Joint Technical Consultation on Avian Influenza at the Human Animal Interface described here was the first opportunity for a large group of influenza experts from the animal and public health sectors to gather and discuss purely technical topics of joint interest that exist at the human-animal interface. During the consultation, three influenza-specific sessions aimed to (1) identify virological characteristics of avian influenza viruses (AIVs) important for zoonotic and pandemic disease, (2) evaluate the factors affecting evolution and emergence of a pandemic influenza strain and identify existing monitoring systems, and (3) identify modes of transmission and exposure sources for human zoonotic influenza infection (including discussion of specific exposure risks by affected countries). A final session was held to discuss broadening the use of tools and systems to other emerging zoonotic diseases. The meeting was structured as short technical presentations with substantial time available for facilitated discussion, to take advantage of the vast influenza knowledge and experience available from the invited expert participants. Particularly important was the identification of gaps in knowledge that have not yet been filled by either sector. Technical discussions focused on H5N1, but included other potentially zoonotic avian and animal influenza viruses whenever possible. During the consultation, the significant threat posed by subtypes other than H5N1 was continually emphasized in a variety of contexts. It was stressed that epidemiological and virological surveillance for these other viruses should be broadening and strengthened. The important role of live bird markets (LBMs) in amplifying and sustaining AIVs in some countries was also a recurring topic, and the need for better understanding of the role of LBMs in human zoonotic exposure and infection was noted. Much is understood about the contribution of various virus mutations and gene combinations to transmissibility, infectivity, and pathogenicity, although it was agreed that the specific constellation of gene types and mutations that would characterize a potentially pandemic virus remains unclear. The question of why only certain humans have become infected with H5N1 in the face of massive exposure in some communities was frequently raised during discussion of human exposure risks. It was suggested that individual-level factors may play a role. More research is needed to address this as well as questions of mode of transmission, behaviors associated with increased risk, virological and ecological aspects, and viral persistence in the environment in order to better elucidate specific human exposure risks. It became clear that great strides have been made in recent years in collaboration between the animal health and public health sectors, especially at the global level. In some countries outbreaks of H5N1 are being investigated jointly. Even greater transparency, cooperation, and information and materials exchange would allow more timely and effective responses in emergency situations, as well as in assessment and planning phases. Ensuring sustainability was also frequently emphasized, e.g. in infrastructure and capacity development and in development of tools and systems for surveillance, assessment and response. It was suggested that one way for tools and systems built or planned to address avian influenza to become more sustainable would be to make them applicable for a broader array of existing and emerging zoonotic diseases.

BACKGROUND

There has been a dramatic increase in the amount of quantitative data derived from the measurement of changes at different levels of biological complexity during the post-genomic era. However, there are a number of issues associated with the use of computational tools employed for the analysis of such data. For example, computational tools such as R and MATLAB require prior knowledge of their programming languages in order to implement statistical analyses on data. Combining two or more tools in an analysis may also be problematic since data may have to be manually copied and pasted between separate user interfaces for each tool. Furthermore, this transfer of data may require a reconciliation step in order for there to be interoperability between computational tools.

RESULTS

Developments in the Taverna workflow system have enabled pipelines to be constructed and enacted for generic and ad hoc analyses of quantitative data. Here, we present an example of such a workflow involving the statistical identification of differentially-expressed genes from microarray data followed by the annotation of their relationships to cellular processes. This workflow makes use of customised maxdBrowse web services, a system that allows Taverna to query and retrieve gene expression data from the maxdLoad2 microarray database. These data are then analysed by R to identify differentially-expressed genes using the Taverna RShell processor which has been developed for invoking this tool when it has been deployed as a service using the RServe library. In addition, the workflow uses Beanshell scripts to reconcile mismatches of data between services as well as to implement a form of user interaction for selecting subsets of microarray data for analysis as part of the workflow execution. A new plugin system in the Taverna software architecture is demonstrated by the use of renderers for displaying PDF files and CSV formatted data within the Taverna workbench.

CONCLUSIONS

Taverna can be used by data analysis experts as a generic tool for composing ad hoc analyses of quantitative data by combining the use of scripts written in the R programming language with tools exposed as services in workflows. When these workflows are shared with colleagues and the wider scientific community, they provide an approach for other scientists wanting to use tools such as R without having to learn the corresponding programming language to analyse their own data.

CONCLUSIONS

MetaFluxNet is a program package for managing information on the metabolic reaction network and for quantitatively analyzing metabolic fluxes in an interactive and customized way. It allows users to interpret and examine metabolic behavior in response to genetic and/or environmental modifications. As a result, quantitative in silico simulations of metabolic pathways can be carried out to understand the metabolic status and to design the metabolic engineering strategies. The main features of the program include a well-developed model construction environment, user-friendly interface for metabolic flux analysis (MFA), comparative MFA of strains having different genotypes under various environmental conditions, and automated pathway layout creation.

BACKGROUND

http://mbel.kaist.ac.kr/

BACKGROUND

A manual for MetaFluxNet is available as PDF file.

Treatment planning based on probability distribution function (PDF) of patient geometries has been shown a potential off-line strategy to incorporate organ motion, but the application of such approach highly depends upon the reproducibility of the PDF. In this paper, we investigated the dependences of the PDF reproducibility on the imaging acquisition parameters, specifically the scan time and the frame rate. Three healthy subjects underwent a continuous 5 min magnetic resonance (MR) scan in the sagittal plane with a frame rate of approximately 10 f s-1, and the experiments were repeated with an interval of 2 to 3 weeks. A total of nine pulmonary vessels from different lung regions (upper, middle and lower) were tracked and the dependences of their displacement PDF reproducibility were evaluated as a function of scan time and frame rate. As results, the PDF reproducibility error decreased with prolonged scans and appeared to approach equilibrium state in subjects 2 and 3 within the 5 min scan. The PDF accuracy increased in the power function with the increase of frame rate; however, the PDF reproducibility showed less sensitivity to frame rate presumably due to the randomness of breathing which dominates the effects. As the key component of the PDF-based treatment planning, the reproducibility of the PDF affects the dosimetric accuracy substantially. This study provides a reference for acquiring MR-based PDF of structures in the lung.

CONCLUSIONS

The XML-based Systems Biology Markup Language (SBML) has emerged as a standard for storage, communication and interchange of models in systems biology. As a machine-readable format XML is difficult for humans to read and understand. Many tools are available that visualize the reaction pathways stored in SBML files, but many components, e.g. unit declarations, complex kinetic equations or links to MIRIAM resources, are often not made visible in these diagrams. For a broader understanding of the models, support in scientific writing and error detection, a human-readable report of the complete model is needed. We present SBML2L(A)T(E)X, a Java-based stand-alone program to fill this gap. A convenient web service allows users to directly convert SBML to various formats, including DVI, L(A)T(E)X and PDF, and provides many settings for customization.

BACKGROUND

Source code, documentation and a web service are freely available at (http://www.ra.cs.uni-tuebingen.de/software/SBML2LaTeX).

This article presents the WHO recommendations on the use of vaccines against tick-borne encephalitis excerpted from the recently published Vaccines against tick-borne encephalitis: WHO position paper. This is the first WHO position paper on the use of tick-borne encephalitis. It was published in the Weekly Epidemiological Record in June 2011. In this paper, footnotes provide a limited number of core references including references to grading tables that assess the quality of scientific evidence for a few key conclusions; a more comprehensive list of references is offered in the Background document on vaccines and vaccination against tick-borne encephalitis available at http://www.who.int/immunization/sage/6_TBE_backgr_18_Mar_net_apr_2011.pdf. In accordance with its mandate to provide guidance to Member States on health policy matters, WHO issues a series of regularly updated position papers on vaccines and combinations of vaccines against diseases that have an international public health impact. These papers are concerned primarily with the use of vaccines in large-scale immunization programmes; they summarize essential background information on diseases and vaccines, and conclude with WHO's current position on the use of vaccines in the global context. This paper reflects the recommendations of WHO's Strategic Advisory Group of Experts (SAGE) on immunization. These recommendations were discussed by SAGE at its April 2011 meeting. Evidence presented at the meeting can be accessed at http://www.who.int/immunization/sage/previous/en/index.html.

Neuropeptides have widespread effects on behavior, but how these molecules alter the activity of their target cells is poorly understood. We employed a new model system in Drosophila melanogaster to assess the electrophysiological and molecular effects of neuropeptides, recording in situ from larval motor neurons, which transgenically express a receptor of choice. We focused on two neuropeptides, pigment-dispersing factor (PDF) and small neuropeptide F (sNPF), which play important roles in sleep/rhythms and feeding/metabolism. PDF treatment depolarized motor neurons expressing the PDF receptor (PDFR), increasing excitability. sNPF treatment had the opposite effect, hyperpolarizing neurons expressing the sNPF receptor (sNPFR). Live optical imaging using a genetically encoded fluorescence resonance energy transfer (FRET)-based sensor for cyclic AMP (cAMP) showed that PDF induced a large increase in cAMP, whereas sNPF caused a small but significant decrease in cAMP. Coexpression of pertussis toxin or RNAi interference to disrupt the G-protein Gαo blocked the electrophysiological responses to sNPF, showing that sNPFR acts via Gαo signaling. Using a fluorescent sensor for intracellular calcium, we observed that sNPF-induced hyperpolarization blocked spontaneous waves of activity propagating along the ventral nerve cord, demonstrating that the electrical effects of sNPF can cause profound changes in natural network activity in the brain. This new model system provides a platform for mechanistic analysis of how neuropeptides can affect target cells at the electrical and molecular level, allowing for predictions of how they regulate brain circuits that control behaviors such as sleep and feeding.

Continued emergence of antimicrobial resistances among gram-positive pathogens requires further development of compounds with novel modes of action. The peptide deformylase inhibitor NVP PDF-713 was tested against 1,837 recent strains of Gram-positive organisms. All NVP PDF-713 MICs were at < or = 4microg/mL except for 6 enterococci (0.3% of strains overall). NVP PDF-713 MIC(90) results were: Staphylococcus aureus, beta-haemolytic and viridans group streptococci and Streptococcus bovis at 1 microg/ml; coagulase-negative staphylococci, Streptococcus pneumoniae, and Listeria spp. at 2 microg/mL; and the enterococci at 4 microg/mL. NVP PDF-713 appears to be a promising new agent worthy of continued in vivo development.

Real-time prostate tracking during intensity-modulated arc radiotherapy requires a reliable prostate position signal during treatment. Many modern linear accelerators have a single gantry-mounted x-ray imager that could be used for intrafraction imaging of implanted prostate markers. The aim of this study was to develop a method to use such a single x-ray imager to estimate the three-dimensional (3D) prostate position in real time during arc treatment delivery and quantify the accuracy of this method in simulations based on 548 prostate trajectories for 17 patients measured with electromagnetic transponders. Imaging at 0.5, 1, 2 and 5 Hz during 360 degrees arc treatments of 1, 2 and 3 min duration was simulated by projecting the prostate position onto the rotating imager. When an image was acquired, a Gaussian probability density function (PDF) for the prostate position was first estimated by maximum likelihood optimization from the set of images acquired so far and then used to estimate the 3D prostate position from the projected position in the image. Since this method needed a PDF right from the onset of the treatment, an initial PDF was obtained with a series of pre-treatment images acquired in 10 s, 20 s or 30 s during a gantry rotation of 60 degrees , 120 degrees or 180 degrees . The accuracy of the estimations was quantified by calculating the root-mean-square (RMS) estimation error for each simulated treatment. The 3D RMS estimation error had a mean value of 0.22 mm and exceeded 1 mm in 0.8% of the cases for 1 min treatments with 5 Hz imaging and 20 s pre-treatment imaging. The position estimation accuracy degraded slightly with reduced imaging frequency or reduced pre-treatment imaging duration. Prolonged treatment duration of 2 and 3 min increased the mean 3D RMS errors to 0.27 mm and 0.30 mm, respectively. The single-imager trajectory estimation method would allow image-guided real-time prostate tracking based on standard equipment for modern linear accelerators.

A voltage-sensitive K+ channel with characteristics of the delayed rectifier was studied in NG108-15 cells using the cell-attached patch-clamp technique. The primary conductance of the channel was 18 pS, but occasional openings to a subconductance state were observed. The average latency to first opening of the channel was about 4 msec. Based on about 20,000 channel openings, the open time probability density function (pdf) required at least three exponentials (time constants of about 0.2, 3 and 9 msec) to achieve an adequate fit to the data. The closed time pdf required at least six exponentials to describe the data (time constants ranging from 0.093 to 440 msec). Thus, the channel exists in at least three open and six closed states. The ensemble average describing the inactivation of the channel was well fit by two exponentials with time constants of 170 msec and 4.2 sec. To examine the effect of changes in membrane lipid composition on the properties of the channel, the phospholipids of the cells were enriched with polyunsaturated fatty acids. In patches from 20:4-enriched cells the conductance, mean first latency, and open-time pdf were similar to control cells. However, the open state probability was increased from 0.25 to 0.44 and the mean closed time was decreased from 20 to 9 msec. The closed time pdf exhibited a higher proportion of closing events associated with short time constants, i.e., the probability of the channel closing into a long-lived closed state was decreased. The decay phase of the ensemble average also was changed; the proportion of the curve described by the slower time constant was almost doubled. Thus, the delayed rectifier from NG108-15 cells can exist in at least three open and six closed states, and changes in membrane lipid composition may have subtle effects on the gating kinetics of the channel.

BACKGROUND

Current medical therapeutic strategies for refractory congestive heart failure (CHF) in the population of 65 years and older with contraindications for heart transplantation are limited. Peritoneal dialysis applied to CHF patients with or without renal impairment showed clinical functional improvement.

METHODS

A single centre, prospective but non-randomized study in 20 patients with severe congestive heart failure refractory to optimal pharmacological therapy [New York Heart Association (NYHA), class IV] was performed between 2000 and 2003. The mean age was 65.71+/-7.66 years. The patients had a baseline glomerular filtration rate of 14.84+/-3.8 ml/min. Fifteen patients were diabetics (type I, 10; type II, five). For all patients, the baseline ejection fraction was <35% (31.2+/-4.7%). The mean Charlson's co-morbidity index was 7.8+/-1.8. Patients were treated initially by 2-5 sessions of continuous veno-venous haemofiltration (CVVH) or sequential haemofiltration (SHF). Automated peritoneal dialysis (APD) was started after implantation of a Tenckhoff catheter. Three APD sessions/week (8 h each), with 15-20 l of dialysis fluid (PDF) per session (10.35+/-3.05 l of 1.5% lactated glucose and 8.95 +/-2.95 l of 4.25% glucose PDF), were performed. Total follow-up ranged between 7 and 35 months (mean 19.80+/-7.37).

RESULTS

After 1 year of follow-up, all patients showed haemodynamic improvement: significant improvement of left cardiac work index (2.33+/-0.69 to 2.59+/-0.47 kg min/m(2)), reduction of the systolic times ratio (61.14+/-12.57 to 39.18+/-13.44%), lower thoracic fluid contents (0.04+/-0.005 to 0.003+/-0.0001 Omega) as well as a regression from NYHA class IV to class I. Need for hospitalization for CHF decreased from 157 to 13 days.

CONCLUSIONS

Peritoneal dialysis appears to be a promising therapeutic tool for patients affected by refractory CHF. Clinical improvement of cardiac function may be related to clearing blood from middle molecular weight myocardial depressant substances, including atrial natriuretic peptide. Prospective multicentre trials are needed to confirm these encouraging results.

BACKGROUND

Transcriptional feedback loops are central to circadian clock function. However, the role of neural activity and membrane events in molecular rhythms in the fruit fly Drosophila is unclear. To address this question, we expressed a temperature-sensitive, dominant negative allele of the fly homolog of dynamin called shibire(ts1) (shi(ts1)), an active component in membrane vesicle scission.

RESULTS

Broad expression in clock cells resulted in unexpectedly long, robust periods (>28 hours) comparable to perturbation of core clock components, suggesting an unappreciated role of membrane dynamics in setting period. Expression in the pacemaker lateral ventral neurons (LNv) was necessary and sufficient for this effect. Manipulation of other endocytic components exacerbated shi(ts1)'s behavioral effects, suggesting its mechanism is specific to endocytic regulation. PKA overexpression rescued period effects suggesting shi(ts1) may downregulate PKA pathways. Levels of the clock component PERIOD were reduced in the shi(ts1)-expressing pacemaker small LNv of flies held at a fully restrictive temperature (29 degrees C). Less restrictive conditions (25 degrees C) delayed cycling proportional to observed behavioral changes. Levels of the neuropeptide PIGMENT-DISPERSING FACTOR (PDF), the only known LNv neurotransmitter, were also reduced, but PERIOD cycling was still delayed in flies lacking PDF, implicating a PDF-independent process. Further, shi(ts1) expression in the eye also results in reduced PER protein and per and vri transcript levels, suggesting that shibire-dependent signaling extends to peripheral clocks. The level of nuclear CLK, transcriptional activator of many core clock genes, is also reduced in shi(ts1) flies, and Clk overexpression suppresses the period-altering effects of shi(ts1).

CONCLUSIONS

We propose that membrane protein turnover through endocytic regulation of PKA pathways modulates the core clock by altering CLK levels and/or activity. These results suggest an important role for membrane scission in setting circadian period.

The National Toxicology Program (NTP) Center for the Evaluation of Risks to Human Reproduction (CERHR) conducted an evaluation of the potential for butyl benzyl phthalate (BBP) to cause adverse effects on reproduction and development in humans. BBP is one of 7 phthalate chemicals evaluated by the NTP CERHR Phthalates Expert Panel. These phthalates were selected for evaluation because of high production volume, extent of human exposures, use in children's products, and/or published evidence of reproductive or developmental toxicity. BBP is used in the production of vinyl tiles and polyvinyl chloride (PVC) to make products such as food conveyor belts, carpet tile, tarps, artificial leather, automotive trim, and traffic cones. The results of this evaluation on BBP are published in a NTP-CERHR monograph which includes: 1) the NTP Brief, 2) the Expert Panel Report on the Reproductive and Developmental Toxicity of Butyl Benzyl Phthalate, and 3) public comments received on the Expert Panel Report. As stated in the NTP Brief, the NTP reached the following conclusions regarding the possible effects of exposure to BBP on human development and reproduction. First, the NTP concludes there is negligible concern for adverse reproductive effects in exposed men. Data are insufficient to reach a conclusion on possible eproductive effects in exposed women. There is no direct evidence that exposure of people to BBP adversely affects reproduction or development, but studies reviewed by the expert panel show that oral exposure of laboratory animals to high doses >>/=1000 mg/kg body weight/day) of BBP can adversely affect development, including development of the male reproductive tract. Second, the NTP concludes that there is minimal concern for developmental effects in fetuses and children. Evidence showed adverse reproductive effects in rats at doses of 100 mg/kg body weight/day, but not at 20 mg/kg/day. Exposure estimates for women of reproductive age were estimated to be 7.8 mug/kg body weight/day. Therefore, this estimated exposure is at least 2,500-to 25,000- fold lower than the toxic dose in rats. NTP-CERHR monographs are transmitted to federal and state agencies, interested parties, and the public and are available in electronic PDF format on the CERHR web site (http://cerhr.niehs.nih.gov) and in printed text or CD-ROM from the CERHR (National Institute of Environmental Health Sciences, P.O. Box 12233, MD EC-32, Research Triangle Park, NC; fax: 919-316-4511).

The R(r):standard allele in maize, which conditions anthocyanin pigmentation in plant and seed tissues in the presence of appropriate complementary factors, is associated with a tandem duplication. The proximal member of the duplication carries P, the plant pigmenting determiner and the distal member member carries S, the seed pigmenting determiner. Derivatives from R(r) that have lost S function are designated r(r). They represent either losses of the distal member of the duplication (P derivatives) or mutations of S to s (P s). Derivatives that have lost P function are designated R(g), and represent either losses of the proximal member of the duplication (S derivatives) or mutations of P to p (p S).-All four possible types of r(r)/R(g) heterozygotes were tested for their capacity to yield R(r) reconstitution by crossing over. No R(r) derivatives were obtained from P/S heterozygotes, a result consistent with the view that P and S occupy corresponding positions in homologous chromosome segments. R(r) reconstitution was detected in both tandem duplication heterozygotes P s/S and P/p S, and was found to be about ten times more frequent in the latter. The ratio of R(r) reconstitution in the two heterozygotes is a function of position of the anthocyanin marker within the duplicated segment. The data from these heterozygotes allow one to measure the distance between P and S, that is to say, the genetic length of the duplicated segment. This distance was found to be 0.16 map units. The highest frequency of R(r) reconstitution was obtained from P s/p S heterozygotes, since direct pairing (see PDF) as well as the p//s type of displaced pairing have the potential to produce R(r) derivatives. One of the R(g) derivatives used in this study, R(g) (6), was found to back-mutate in some sublines to R(r). The basis for this instability remains unknown.

OBJECTIVE

To determine the prevalence of depressive and high trait anxiety symptoms and substance use, including alcohol and nicotine, in first-year and second-year medical students in Skopje University Medical School, Republic of Macedonia.

BACKGROUND

It is important to investigate medical students because they are under significant pressure during early years of medical education, a period during which the attitudes and behaviors of physicians develop.

METHODS

A cross-sectional survey in classroom settings, using an anonymous self-administered questionnaire, was performed in 354 participants (181 first-year, 118 females and 63 males and 173 second-year medical students, 116 females and 57 males) aged 18 to 23 years. The Beck Depression Inventory (BDI) and Taylor Manifest Anxiety Scale (TMAS) were used to determine depressive and high trait anxiety symptoms. BDI scores 17 or higher were categorized as depressive and TMAS scores 16 or higher as high anxiety symptoms. A Student t-test was used for continuous data analysis.

RESULTS

Out of all participants 10.4% had BDI score 17 or higher and 65.5% had TMAS score 16 or higher. Alcohol was the most frequently used substance in both groups. Smoking prevalence was 25%. Benzodiazepines (diazepam, alprazolam) use was 13.1%. Illicit drug use was rare (1.1% in freshmen and 3.6% in juniors) in both groups.

CONCLUSIONS

High frequency of manifest high anxiety symptoms and depressive symptoms and benzodiazepine use among Macedonian junior medical students should be taken seriously and a student counseling service offering mental health assistance is necessary (Tab. 3, Ref. 23). Full Text (Free, PDF) www.bmj.sk.

We report the results of the international daptomycin surveillance programs for Europe, Latin America, and selected Asia-Pacific nations. A total of 7948 consecutive Gram-positive organisms of clinical significance were collected in 2011 and susceptibility tested against daptomycin and various comparator agents by Clinical and Laboratory Standards Institute (Clinical and Laboratory Standards Institute. M07-A9. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard: ninth edition Wayne, PA: CLSI. 2012.; Cubicin Package Insert 2012. Cubist Pharmaceuticals, Inc, Lexington, MA. Available at http://www.cubicin.com/pdf/PrescribingInformation.pdf. Accessed January 1, 2012.) broth microdilution methods. The test medium was adjusted to contain physiological levels of calcium (50 mg/L) when testing daptomycin. Daptomycin exhibited potent activity against methicillin-susceptible and -resistant Staphylococcus aureus overall and for each region (MIC(50/90), 0.25-0.5/0.5 μg/mL), with susceptibility rates at 100.0% in Latin America, Australia/New Zealand, and India, and at 99.9% in Europe. The daptomycin MIC(50/90) for coagulase-negative staphylococci was also at 0.25-0.5/0.5 μg/mL, and only 1 isolate was considered nonsusceptible with a MIC value at 2 μg/mL. Daptomycin was also highly active against Enterococcus faecalis (MIC(50/90), 1/1-2 μg/mL) and E. faecium (MIC(50/90), 2/2 μg/mL for both vancomycin-susceptible and -resistant isolates). All enterococcal isolates were susceptible to daptomycin (MIC, ≤4 μg/mL) and tigecycline. Susceptibility to linezolid for E. faecalis was at 100.0%, while for E. faecium regional susceptibility rates were at 100.0% except in Europe (99.0%). Viridans group streptococci (MIC(50/90), 0.25/1 μg/mL) and β-haemolytic streptococci (MIC(50/90), ≤0.06/0.25 μg/mL) continue to be very susceptible to daptomycin. In summary, the results of this investigation document the high potency and wide spectrum of daptomycin when tested against a large resistance-surveillance collection of Gram-positive pathogens and indicate that daptomycin nonsusceptibility remains rare among indicated species after many years of clinical use worldwide.

Endogenous circadian clocks are inherent to all living organisms. They are needed to guarantee successful life since they regulate very important biological processes such as behavior and reproduction. Secretin-like G-protein coupled receptors are very important factors in the signal transduction pathways of circadian clocks. In this review, we will focus on the role of two secretin-like signaling pathways that play an important role in the regulation of the mammalian and the insect clock, respectively: the pituitary adenylate cyclase-activating polypeptide (PACAP) and pigment dispersing factor (PDF) signaling pathways. Both pathways are most likely related although their function in the biological clock differs.

To clarify the demographic and clinicolaboratory features of postdialysis fatigue (PDF), we enrolled 85 patients on maintenance hemodialysis in a cross-sectional study using validated questionnaires and chart review. Forty-three patients complained of fatigue after dialysis. On formal testing using the Kidney Disease Questionnaire, the PDF group had statistically greater severity of fatigue and somatic complaints than the group of patients without subjective fatigue (P = 0.03 and 0.04, respectively). On a scale measuring intensity of fatigue (1 = least to 5 = worst), the PDF group average was 3.4 +/- 1.2. PDF subjects reported that 80% +/- 25% of dialysis treatments were followed by fatigue symptoms. In 28 (65%) of patients, the symptoms started with the first dialysis treatment. They reported needing an average of 4.8 hours of rest or sleep to overcome the fatigue symptoms (range, 0 to 24 hours). There were no significant differences between patients with and without PDF in the following parameters: age; sex; type of renal disease; presence of diabetes mellitus, heart disease (congestive, ischemic), or chronic obstructive lung disease; blood pressure response to dialysis; type or adequacy of dialysis regimen; hematocrit; electrolytes; blood urea nitrogen; creatinine; cholesterol; albumin; parathyroid hormone; ejection fraction; and use of antihistamines, benzodiazepines, and narcotics. In the fatigue group, there was significantly greater use of antihypertensive medications known to have fatigue as a side effect (P = 0.007). Depression was more common in the fatigue group by Beck Depression score (11.6 +/- 8.0 v 7.8 +/- 6.3; P = 0.02). We conclude that (1) postdialysis fatigue is a common, often incapacitating symptom in patients on chronic extracorporeal dialysis; (2) no routinely measured parameter of clinical or dialytic function appears to predict postdialysis fatigue; and (3) depression is highly associated with postdialysis fatigue, but the cause-effect relationship is unclear.

OBJECTIVE

Although peritoneal dialysis (PD) is a widely accepted form of renal replacement therapy, concerns remain regarding the bioincompatible nature of standard PD fluid (PDF). Short-term studies of new biocompatible PDFs low in glucose degradation products (GDPs) reveal divergent results with respect to peritoneal integrity.

METHODS

We studied 125 patients on maintenance PD who were assigned, by simple randomization, to receive either conventional or low-GDP PDF at PD initiation. Parameters of dialysis adequacy and peritoneal transport of small solutes were determined at initiation and after a period of maintenance PD at the time when serum and overnight effluent dialysate were simultaneously collected and assayed for various cytokines, chemokines, adipokines, and cardiac biomarkers. All patients were further followed prospectively for an average of 15 months from the day of serum and effluent collection to determine patient survival and cardiovascular events.

RESULTS

Patients treated with conventional or low-GDP PDF were matched for sex, age, duration of dialysis, dialysis adequacy, and incidence of cardiovascular disease or diabetes. After an average of 2.3 years of PD treatment, the weekly total and peritoneal creatinine clearance, and the total and peritoneal Kt/V were comparable in the groups. However, urine output was higher in patients using low-GDP PDF despite there having been no difference between the groups at PD initiation. Patients using low-GDP PDF also experienced a slower rate of decline of residual glomerular filtration and urine output than did patients on conventional PDF. Compared with serum concentrations, effluent concentrations of tumor necrosis factor α, hepatocyte growth factor, macrophage migration inhibitory factor, interleukins 8 and 6, C-reactive protein, and leptin were found to be higher in both groups of patients after long-term PD, suggesting that the peritoneal cavity was the major source of those mediators. Compared with patients on low-GDP PDF, patients on conventional fluid showed elevated leptin and reduced adiponectin levels in serum and effluent. The effluent concentration of interleukin 8 was significantly lower in patients using low-GDP PDF. The survival rate and incidence of cardiovascular complications did not differ between these groups after maintenance PD for an average of 3.6 years.

CONCLUSIONS

It appears that low-GDP PDF results in an improvement of local peritoneal homeostasis through a reduction of chronic inflammatory status in the peritoneum.

A variety of cell mutants were obtained by a single 5'-bromodeoxyuridine (BrdU) treatment of an nonproducer (NP) cell line transformed by the Kirsten strain of murine sarcoma virus (Ki-MSV). Isolation procedures of these cell See PDF for Structure mutants are described. The cell mutants obtained were classified by tumorigenic potential and shedding of Type C virus particles. The cell mutants were classified into four groups: (A) tumorigenic, without particles; (B) tumorigenic, with Type C particles; (C) nontumorigenic, without particles; and (D) nontumorigenic, with Type C particles. The tumorigenic cell lines showed variability in morphology with both flat and typical transformed appearing cell lines showing equal transplantability.

The K-line, which is a virtual line that connects the midpoints of the anteroposterior diameter of the spinal canal at C2 and C7 in a plain lateral radiogram, is a useful preoperative predictive indicator for sufficient decompression by laminoplasty (LMP) for ossification of the posterior longitudinal ligament (OPLL). K-line is defined as (+) when the peak of OPLL does not exceed the K-line, and is defined as (-) when the peak of OPLL exceeds the K-line. For patients with K-line (-) OPLL, LMP often results in poor outcome. The aim of the present study was to compare the clinical outcome of LMP, posterior decompression with instrumented fusion (PDF) and anterior decompression and fusion (ADF) for patients with K-line (-) OPLL.

The present study included patients who underwent surgical treatment including LMP, PDF and ADF for K-line (-) cervical OPLL. We retrospectively compared the clinical outcome of those patients in terms of Japanese Orthopedic Association score (JOA score) recovery rate.

JOA score recovery rate was significantly higher in the ADF group compared with that in the LMP group and the PDF group. The JOA score recovery rate in the PDF group was significantly higher than that in the LMP group.

LMP should not be used for K-line (-) cervical OPLL. ADF is one of the suitable surgical treatments for K-line (-) OPLL. Both ADF and PDF are applicable for K-line (-) OPLL according to indications set by each institute and surgical decisions.

The substrate specificity of Escherichia coli peptide deformylase was investigated by measuring the efficiency of the enzyme to cleave formyl- peptides of the general formula Fo-Xaa-Yaa-NH2, where Xaa represents a set of 27 natural and unusual amino acids and Yaa corresponds to a set of 19 natural amino acids. Substrates with bulky hydrophobic side-chains at the P1' position were the most efficiently cleaved, with catalytic efficiencies greater by two to five orders of magnitude than those associated with polar or charged amino acid side-chains. Among hydrophobic side-chains, linear alkyl groups were preferred at the P1' position, as compared to aryl-alkyl side-chains. Interestingly, in the linear alkyl substituent series, with the exception of norleucine, deformylase exhibits a preference for the substrate containing Met in the P1' position. Next, the influence in catalysis of the second side-chain was studied after synthesis of 20 compounds of the formula Fo-Nle-Yaa-NH2. Their deformylation rates varied within a range of only one order of magnitude. A 3D model of the interaction of PDF with an inhibitor was then constructed and revealed indeed the occurrence of a deep and hydrophobic S1' pocket as well as the absence of a true S2' pocket. These analyses pointed out a set of possible interactions between deformylase and its substrates, which could be the ground driving substrate specificity. The validity of this enzyme:substrate docking was further probed with the help of a set of site-directed variants of the enzyme. From this, the importance of residues at the bottom of the S1' pocket (Ile128 and Leu125) as well as the hydrogen bond network that the main chain of the substrate makes with the enzyme were revealed. Based on the numerous homologies that deformylase displays with thermolysin and metzincins, a mechanism of enzyme:substrate recognition and hydrolysis could finally be proposed. Specific features of PDF with respect to other members of the enzymes with motif HEXXH are discussed.