Ventilatory function as a predictor of fatal stroke.
To investigate the relation between ventilatory function and subsequent mortality due to cerebrovascular disease. Prospective longitudinal study. A total of 18,403 male civil servants aged 40-64 years at entry examination for the Whitehall study. Mortality from cerebrovascular disease (ICD8 430-438) after 18 years of follow up. In all, 262 men with sinus rhythm at entry died due to stroke during the 18 years of follow up. Compared with men with a forced expiratory volume in one second of greater than or equal to 3.5 litres those with a value of less than 3.0 litres were almost twice as likely to die of cerebrovascular disease (rate ratio adjusted for age and systolic blood pressure = 1.88, 95% confidence interval 1.32 to 2.69). This increased risk occurred within each tertile of systolic blood pressure. Nested case-control analyses were used to control precisely for confounding effects of age, height, and smoking (by matching) and employment grade and physiological risk factors (by modelling). The effect of forced expiratory volume in one second was independent of age, height, smoking habits, employment grade, blood pressure, weight, cholesterol concentration, glucose tolerance, electrocardiographic abnormalities, history of chest pain, and history of intermittent claudication. Measurements of ventilatory function may assist clinical decisions about whether to treat mild hypertension. Impaired ventilatory function and stroke may share common causes.


Residues of human cytomegalovirus DNA polymerase catalytic subunit UL54 that are necessary and sufficient for interaction with the accessory protein UL44.
The human cytomegalovirus DNA polymerase contains a catalytic subunit, UL54, and an accessory protein, UL44. Recent studies suggested that UL54 might interact via its extreme C terminus with UL44 (A. Loregian, R. Rigatti, M. Murphy, E. Schievano, G. Palu', and H. S. Marsden, J. Virol. 77:8336-8344, 2003). To address this hypothesis, we quantitatively measured the binding of peptides corresponding to the extreme C terminus of UL54 to UL44 by using isothermal titration calorimetry. A peptide corresponding to the last 22 residues of UL54 was sufficient to bind specifically to UL44 in a 1:1 complex with a dissociation constant of ca. 0.7 microM. To define individual residues in this segment that are crucial for interacting with UL44, we engineered a series of mutations in the C-terminal region of UL54. The UL54 mutants were tested for their ability to interact with UL44 by glutathione S-transferase pulldown assays, for basal DNA polymerase activity, and for long-chain DNA synthesis in the presence of UL44. We observed that deletion of the C-terminal segment or substitution of alanine for Leu1227 or Phe1231 in UL54 greatly impaired both the UL54-UL44 interaction in pulldown assays and long-chain DNA synthesis without affecting basal polymerase activity, identifying these residues as important for subunit interaction. Thus, like the herpes simplex virus UL30-UL42 interaction, a few specific side chains in the C terminus of UL54 are crucial for UL54-UL44 interaction. However, the UL54 residues important for interaction with UL44 are hydrophobic and not basic. This information might aid in the rational design of new drugs for the treatment of human cytomegalovirus infection.


Local Arp2/3-dependent actin assembly modulates applied traction force during apCAM adhesion site maturation.
Homophilic binding of immunoglobulin superfamily molecules such as the Aplysia cell adhesion molecule (apCAM) leads to actin filament assembly near nascent adhesion sites. Such actin assembly can generate significant localized forces that have not been characterized in the larger context of axon growth and guidance. We used apCAM-coated bead substrates applied to the surface of neuronal growth cones to characterize the development of forces evoked by varying stiffness of mechanical restraint. Unrestrained bead propulsion matched or exceeded rates of retrograde network flow and was dependent on Arp2/3 complex activity. Analysis of growth cone forces applied to beads at low stiffness of restraint revealed switching between two states: frictional coupling to retrograde flow and Arp2/3-dependent propulsion. Stiff mechanical restraint led to formation of an extensive actin cup matching the geometric profile of the bead target and forward growth cone translocation; pharmacological inhibition of the Arp2/3 complex or Rac attenuated F-actin assembly near bead binding sites, decreased the efficacy of growth responses, and blocked accumulation of signaling molecules associated with nascent adhesions. These studies introduce a new model for regulation of traction force in which local actin assembly forces buffer nascent adhesion sites from the mechanical effects of retrograde flow.


The K-complex and slow oscillation in terms of a mean-field cortical model.
We use a mean-field macrocolumn model of the cerebral cortex to offer an interpretation of the K-complex of the electroencephalogram to complement those of more detailed neuron-by-neuron models. We interpret the K-complex as a momentary excursion of the cortex from a stable low-firing state to an unstable high-firing state, and hypothesize that the related slow oscillation can be considered as the periodic oscillation between two meta-stable solutions of the mean-field model. By incorporating a Hebbian-style learning rule that links the growth in synapse strength to fluctuations in soma potential, we demonstrate a self-organization behaviour that draws the modelled cortex close to the edge of stability of the low-firing state. Furthermore, a very slow oscillation can occur in the excitability of the cortex that has similarities with the infra-slow oscillation of sleep.


Measuring the performance of individual physicians by collecting data from multiple health plans: the results of a two-state test.
Quality measurement and reporting have emerged as important tools that providers, health plans, and other stakeholders can use to identify gaps in quality and focus resources on improving care. Yet identifying, measuring, and evaluating the care that physicians and other health care providers deliver is complicated by limited data, privacy concerns, and the challenge of trying to compare data from diverse sources. This article describes an effort to pilot-test in Florida and Colorado a consistent approach to individual physician performance measurement using data compiled from multiple health plans. Our approach could be used as the basis for making comparable performance information available nationwide. Additional efforts are needed to address key issues, including ways to effectively engage providers in the use of performance information.


Endogenous neuroprotection: hamartin modulates an austere approach to staying alive in a recession.
Tuberous sclerosis complex 1 (hamartin) is an effective endogenous neuroprotectant. Understanding the endogenous mechanism for neuroprotection mediated by hamartin may afford a novel approach to effective treatment of neurological diseases such as stroke, neurodegenerative diseases, and epilepsy, with possible applications to nonneurological conditions.


Immunotherapy of multiple myeloma.
Multiple myeloma (MM), a bone marrow-resident hematological malignancy of plasma cells, has remained largely incurable despite dramatic improvements in patient outcomes in the era of myeloma-targeted and immunomodulatory agents. It has recently become clear that T cells from MM patients are able to recognize and eliminate myeloma, although this is subverted in the majority of patients who eventually succumb to progressive disease. T cell exhaustion and a suppressive bone marrow microenvironment have been implicated in disease progression, and once these are established, immunotherapy appears largely ineffective. Autologous stem cell transplantation (ASCT) is a standard of care in eligible patients and results in immune effects beyond cytoreduction, including lymphodepletion, T cell priming via immunogenic cell death, and inflammation; all occur within the context of a disrupted bone marrow microenvironment. Recent studies suggest that ASCT reestablishes immune equilibrium and thus represents a logical platform in which to intervene to prevent immune escape. New immunotherapies based on checkpoint inhibition targeting the immune receptor TIGIT and the deletion of suppressive myeloid populations appear attractive, particularly after ASCT. Finally, the immunologically favorable environment created after ASCT may also represent an opportunity for approaches utilizing bispecific antibodies or chimeric antigen receptor T cells.


Analysis of altered microRNA expression profiles in proximal renal tubular cells in response to calcium oxalate monohydrate crystal adhesion: implications for kidney stone disease.
Calcium oxalate monohydrate (COM) is the major crystalline component in kidney stones and its adhesion to renal tubular cells leads to tubular injury. However, COM-induced toxic effects in renal tubular cells remain ambiguous. MicroRNAs (miRNAs) play an important role in gene regulation at the posttranscriptional levels. The present study aimed to assess the potential changes in microRNAs of proximal renal tubular cells in response to the adhesion of calcium oxalate monohydrate (COM) crystals. Lactate dehydrogenase (LDH) activity and DAPI staining were used to measure the toxic effects of HK-2 cells exposed to COM crystals. MicroRNA microarray and mRNA microarray were applied to evaluate the expression of HK-2 cells exposed to COM crystals. Quantitative real-time PCR (qRT-PCR) technology was used to validate the microarray results. Target prediction, Gene Ontology (GO) analysis and pathway analysis were applied to predict the potential roles of microRNAs in biological processes. Our study showed that COM crystals significantly altered the global expression profile of miRNAs in vitro. After 24 h treatment with a dose (1 mmol/L), 25 miRNAs were differentially expressed with a more than 1.5-fold change, of these miRNAs, 16 were up-regulated and 9 were down-regulated. A majority of these differentially expressed miRNAs were associated with cell death, mitochondrion and metabolic process. Target prediction and GO analysis suggested that these differentially expressed miRNAs potentially targeted many genes which were related to apoptosis, regulation of metabolic process, intracellular signaling cascade, insulin signaling pathway and type 2 diabetes. Our study provides new insights into the role of miRNAs in the pathogenesis associated with nephrolithiasis.


Characterisation of fibrillin-1 cDNA clones in a human fibroblast cell line that assembles microfibrils.
Fibrillin-1 is a large extracellular glycoprotein which is a major structural component of 10-12 nm microfibrils. Defects in human fibrillin-1 give rise to the autosomal dominant connective tissue disease the Marfan syndrome and related disorders. Previous studies examining the biosynthesis and secretion of recombinant fibrillin-1 fragments have been performed in cell lines which do not assemble fibrillin into extracellular 10-12 nm microfibrils. Conflicting data have been obtained regarding N-terminal processing. In this study we have characterised a human fibroblast cell line MSU-1.1 which shows a similar endogenous fibrillin-1 pulse chase profile to primary human dermal fibroblasts and produces microfibrils. Expression of a approximately 50 kDa N-terminal recombinant peptide in MSU-1.1 resulted in efficient secretion of this peptide into conditioned media, N-terminal sequence analysis of the purified peptide identified 2 protease cleavage sites and a presumed signal peptidase site. Together these data identify the natural leader sequence of fibrillin-1 and the presence of two processing sites in the N-terminus of fibrillin-1. The identification of an N-terminal processing site in recombinant fibrillin-1 similar to that obtained in a previous study which used an HT1080 fibrosarcoma host cell line excludes defective N-terminal processing as the cause of the assembly defect in this cell line. A full length normal and mutant fibrillin cDNA (approximately 8.6 kb) was constructed and stable integration of each into MSU1.1 led to RNA transcription at approximately 5% of endogenous levels. This is the first report of transcription from the full length fibrillin-1 cDNA. The low levels of transcription achieved, suggest that additional upstream and downstream DNA sequence elements will be required for high levels of full length fibrillin-1 cDNA expression.


Computed optical interferometric tomography for high-speed volumetric cellular imaging.
Three-dimensional high-resolution imaging methods are important for cellular-level research. Optical coherence microscopy (OCM) is a low-coherence-based interferometry technology for cellular imaging with both high axial and lateral resolution. Using a high-numerical-aperture objective, OCM normally has a shallow depth of field and requires scanning the focus through the entire region of interest to perform volumetric imaging. With a higher-numerical-aperture objective, the image quality of OCM is affected by and more sensitive to aberrations. Interferometric synthetic aperture microscopy (ISAM) and computational adaptive optics (CAO) are computed imaging techniques that overcome the depth-of-field limitation and the effect of optical aberrations in optical coherence tomography (OCT), respectively. In this work we combine OCM with ISAM and CAO to achieve high-speed volumetric cellular imaging. Experimental imaging results of ex vivo human breast tissue, ex vivo mouse brain tissue, in vitro fibroblast cells in 3D scaffolds, and in vivo human skin demonstrate the significant potential of this technique for high-speed volumetric cellular imaging.


Water-only fasting and an exclusively plant foods diet in the management of stage IIIa, low-grade follicular lymphoma.
Follicular lymphoma (FL), the second most common non-Hodgkin's lymphoma (NHL), is well characterised by a classic histological appearance and an indolent course. Current treatment protocols for FL range from close observation to immunotherapy, chemotherapy and/or radiotherapies. We report the case of a 42-year-old woman diagnosed by excisional biopsy with stage IIIa, grade 1 FL. In addition to close observation, the patient underwent a medically supervised, 21-day water-only fast after which enlarged lymph nodes were substantially reduced in size. The patient then consumed a diet of minimally processed plant foods free of added sugar, oil and salt (SOS), and has remained on the diet since leaving the residential facility. At 6 and 9-month follow-up visits, the patient's lymph nodes were non-palpable and she remained asymptomatic. This case establishes a basis for further studies evaluating water-only fasting and a plant foods, SOS-free diet as a treatment protocol for FL.


Design and application of an in vivo reporter assay for phenylalanine ammonia-lyase.
Phenylalanine ammonia-lyase (PAL) is an important enzyme that links primary metabolism to secondary metabolism. Its efficiency is often a critical factor that affects the overall flux of a related metabolic pathway, the titer of the final products, and the efficacy of PAL-based therapies. Thus, PAL is a common target for metabolic engineering, and it is of significant interest to screen efficient PALs for industrial and medical applications. In this study, a novel and efficient visible reporter assay for screening of PAL efficiency in Escherichia coli was established based on a plant type III polyketide biosynthetic pathway. The candidate PALs were co-expressed with a 4-coumarate:CoA ligase 4CL1 from Arabidopsis thaliana and curcuminoid synthase (CUS) from Oryza sativa in E. coli BL21(DE3) to form a dicinnamoylmethane biosynthetic pathway. Taking advantage of the yellow color of the product, a microplate-based assay was designed to measure the titer of dicinnamoylmethane, which was validated by HPLC analysis. The different titers of the product reflect the overall performance (expression level and enzymatic activity) of the individual PALs in E. coli. Using this system, we have screened three PALs (PAL1, PAL3, and PAL4) from Trifolium pratense, among which PAL1 showed the best performance in E. coli. The engineered E. coli strain containing PAL1, 4CL1, and CUS led to the production of dicinnamoylmethane at a high level of 0.36 g/l. Supplement of 2-fluoro-phenylalanine yielded two fluorinated dicinnamoylmethane derivatives, 6,6'-difluoro-dicinnamoylmethane and 6-fluoro-dicinnamoylmethane, of which the latter is a new curcuminoid.


Sexual dysfunction in patients with alcohol and opioid dependence.
There are limited numbers of studies which have evaluated the sexual dysfunction (SD) in patients with alcohol and opioids dependence. This article reviews the existing literature. Electronic searches were carried out using the PubMed, Google Scholar, and ScienceDirect to locate the relevant literature. Subjects addicted to heroin or on methadone maintenance treatment (MMT) or buprenorphine maintenance treatment (BMT) show higher rates of SD in comparison to the general population. SD rates have ranged 34-85% for heroin addicts, 14-81% for MMT, 36-83% for BMT, and 90% for naltrexone maintenance. The rates of SD in alcohol-dependent population have ranged 40-95.2%, with rates being consistently much higher in alcohol-dependent population than in the healthy controls or social drinkers. The common SDs reported have been erectile dysfunction followed by premature ejaculation, retarded ejaculation and decreased sexual desire among men, and dyspareunia and vaginal dryness among women. This review suggests that long-term use of alcohol and opioids are associated with SD in almost all domains of sexual functioning. There is a need to increase the awareness of clinicians about this association as many times SD in patients with substance abuse lead to poor treatment compliance and relapse. Further, there is a need to carry out more number of studies to understand the relationship in a better way.


AgSwe1p regulates mitosis in response to morphogenesis and nutrients in multinucleated Ashbya gossypii cells.
Nuclei in the filamentous, multinucleated fungus Ashbya gossypii divide asynchronously. We have investigated what internal and external signals spatially direct mitosis within these hyphal cells. Mitoses are most common near cortical septin rings found at growing tips and branchpoints. In septin mutants, mitoses are no longer concentrated at branchpoints, suggesting that the septin rings function to locally promote mitosis near new branches. Similarly, cells lacking AgSwe1p kinase (a Wee1 homologue), AgHsl1p (a Nim1-related kinase), and AgMih1p phosphatase (the Cdc25 homologue that likely counteracts AgSwe1p activity) also have mitoses distributed randomly in the hyphae as opposed to at branchpoints. Surprisingly, however, no phosphorylation of the CDK tyrosine 18 residue, the conserved substrate of Swe1p kinases, was detected in normally growing cells. In contrast, abundant CDK tyrosine phosphorylation was apparent in starving cells, resulting in diminished nuclear density. This starvation-induced CDK phosphorylation is AgSwe1p dependent, and overexpressed AgSwe1p is sufficient to delay nuclei even in rich nutrient conditions. In starving cells lacking septins or AgSwe1p negative regulators, the nuclear density is further diminished compared with wild type. We have generated a model in which AgSwe1p may regulate mitosis in response to cell intrinsic morphogenesis cues and external nutrient availability in multinucleated cells.


Addition of intravenous beta(2)-agonists to inhaled beta(2)-agonists for acute asthma.
Inhaled beta-agonist therapy is central to the management of acute asthma. This review evaluates the benefit of an additional use of intravenous beta(2)-agonist agents. To determine the benefit of adding intravenous (IV) beta(2)-agonists to inhaled beta(2)-agonist therapy for acute asthma treated in the emergency department. Randomised controlled trials (RCTs) were identified using the Cochrane Airways Group Register which is a compilation of systematic searches of MEDLINE, EMBASE, CINAHL, and CENTRAL as well as handsearching of 20 respiratory journals. Bibliographies from included studies and known reviews were also searched. Primary authors and content experts were contacted to identify eligible studies. The search was performed in September 2012. Only RCTs were considered for inclusion. Studies were included if patients presented to the emergency department with acute asthma and were treated with IV beta(2)-agonists with inhaled beta(2)-agonist therapy and existing standard treatments versus inhaled beta(2)-agonists and existing standard treatments. Two review authors independently extracted data and confirmed their findings with corresponding authors of trials. We obtained missing data from authors or calculated from data present in the papers. We used fixed-effect model for odds ratios (OR) and for mean differences (MD) we used both fixed-effect and random-effects models and reported 95% confidence intervals (CI). From 109 potentially relevant studies only three (104 patients) met our inclusion criteria: Bogie 2007 (46 children), Browne 1997 (29 children) and Nowak 2010 (29 adults). Bogie 2007 investigated the addition of intravenous terbutaline to high dose nebulised albuterol in children with acute severe asthma, requiring intensive care unit (ICU) admission. Browne 1997 investigated the benefit of adding intravenous salbutamol to inhaled salbutamol in children with acute severe asthma in the emergency department. Nowak 2010 investigated addition of IV bedoradrine to standard care (nebulised albuterol, ipratropium and oral corticosteroids) among adults, and was reported as a conference abstract only.There was no significant advantage (OR 0.29; 95%CI 0.06 to 1.38, one trial, 29 adults) for adding IV bedoradrine to standard care (nebulised albuterol, ipratropium and oral corticosteroids) with regard to hospitalisation rates.Various outcome indicators for the length of stay were reported among the trials. Browne 1997 reported a significantly shorter recovery time (in terms of cessation of 30 minute salbutamol) for children in the IV salbutamol with inhaled salbutamol group (four hours) versus the 11.1 hours for the inhaled salbutamol group (P = 0.03). Time to cessation of hourly nebuliser was also significantly shorter (P = 0.02) for the IV plus inhaled salbutamol group (11.5 hours versus 21.2 hours), and they were ready for emergency patient discharge on average 9.7 hours earlier than the inhaled salbutamol group (P < 0.05). In a paediatric ICU study Bogie 2007 reported no significant advantage in length of paediatric ICU admission (hours) for adding IV terbutaline to nebulised albuterol (MD -12.95, 95% CI: -38.74, 12.84).Browne 1997 reported there were only six out of 14 children with a pulmonary index score above six in the IV plus inhaled salbutamol group at two hours compared with 14 of the 15 in the inhaled salbutamol group (P = 0.02)In Browne 1997 there was a higher proportion of tremor in the IV plus inhaled salbutamol group than in the inhaled salbutamol group (P < 0.02). Nowak 2010 did not report any statistically significant adverse effects associated with adding IV bedoradrine to standard care (nebulised albuterol, ipratropium and oral corticosteroids). Troponin levels were elevated in three children in the IV terbutaline + nebulised albuterol group at 12 and 24 hours in Bogie 2007 There is very limited evidence from one study (Browne 1997) to support the use of IV beta(2)-agonists in children with severe acute asthma with respect to shorter recovery time, and similarly there is limited evidence (again from one study Browne 1997) suggesting benefit with regard to pulmonary index scores; however this advantage needs to be considered carefully in relation to the increased side effects associated with IV beta(2)-agonists. We identified no significant benefits for adults with severe acute asthma. Until more, adequately powered, high quality clinical trials in this area are conducted it is not possible to form a robust evaluation of the addition of IV beta(2)-agonists in children or adults with severe acute asthma.


Genome plasticity favours double chromosomal Tn4401b-blaKPC-2 transposon insertion in the Pseudomonas aeruginosa ST235 clone.
Pseudomonas aeruginosa Sequence Type 235 is a clone that possesses an extraordinary ability to acquire mobile genetic elements and has been associated with the spread of resistance genes, including genes that encode for carbapenemases. Here, we aim to characterize the genetic platforms involved in resistance dissemination in blaKPC-2-positive P. aeruginosa ST235 in Colombia. In a prospective surveillance study of infections in adult patients attended in five ICUs in five distant cities in Colombia, 58 isolates of P. aeruginosa were recovered, of which, 27 (46.6%) were resistant to carbapenems. The molecular analysis showed that 6 (22.2%) and 4 (14.8%) isolates harboured the blaVIM and blaKPC-2 genes, respectively. The four blaKPC-2-positive isolates showed a similar PFGE pulsotype and belonged to ST235. Complete genome sequencing of a representative ST235 isolate shows a unique chromosomal contig of 7097.241 bp with eight different resistance genes identified and five transposons: a Tn6162-like with ant(2″)-Ia, two Tn402-like with ant(3″)-Ia and blaOXA-2 and two Tn4401b with blaKPC-2. All transposons were inserted into the genomic islands. Interestingly, the two Tn4401b copies harbouring blaKPC-2 were adjacently inserted into a new genomic island (PAGI-17) with traces of a replicative transposition process. This double insertion was probably driven by several structural changes within the chromosomal region containing PAGI-17 in the ST235 background. This is the first report of a double Tn4401b chromosomal insertion in P. aeruginosa, just within a new genomic island (PAGI-17). This finding indicates once again the great genomic plasticity of this microorganism.


IMMUNOLOGICAL STUDIES WITH HERPES VIRUS WITH A CONSIDERATION OF THE HERPES-ENCEPHALITIS PROBLEM.
In the preceding experiments observations have been reported upon the nature of herpes virus which confirm the suspicion that the virus is intracellularly located in the infected nervous system. In regard to the immunological conditions existing in this disease, our experiments have reaffirmed that herpes virus can be neutralized with the serum of actively immunized animals and have offered an explanation for the irregularity of the results of others, as well as our own. It has been found that brain extracts possess some virus-neutralizing power, but considerably less than the serum of the corresponding animals. Attempts at passive immunization with neutralizing serum were uniformly negative, even when the serum was introduced into the cisterna magna 12 to 24 hours before infection with the virus. It has been shown that active immunity can be attained only when some degree of reaction to the living virus has occurred. Rabbits which survived neutralized serum-virus mixtures did not acquire immunity nor did those treated with virus phenolized to the extent of actual destruction. This point suggests a reinvestigation of the Semple method of rabies immunization. In so far as our studies touched upon the herpes-encephalitis problem we have uniformly failed in attempts to transfer herpes virus directly from man to rabbits. These results are in contradiction to those of most of the earlier workers, but in keeping with the recent reports of Flexner and Amoss. Attempts to overcome the difficulty of transfer by the recently published technique of Perdrau were unsuccessful. Furthermore, animals repeatedly treated with human encephalitis material, either fresh or glycerolated, as practised in the Perdrau method, failed to acquire the slightest degree of immunity to subsequent herpes inoculation. By the inoculation of very small doses or by infection of partially immunized rabbits, as described above, we have succeeded in modifying the characteristic herpetic syndrome in rabbits in a manner which simulates many of the clinical features of human encephalitis. Our own experience forces the conclusion that no valid proof exists upon which can be based an assertion concerning the identity of the virus of herpes with that of encephalitis lethargica. Either the two viruses are entirely unrelated, or else prolonged sojourn in the central nervous system of man attenuates the virus for rabbits to an extent analogous to that in which rabies virus is attenuated for man by passage through rabbits. The isolated successes of Levaditi and of Doerr and their assistants might thus be regarded as fortunate exceptions in which material incompletely attenuated had been at their disposal. We suggest this point of view as an alternative working hypothesis largely because the results we are reporting, as well as those of Flexner and Amoss, are in flat contradiction to the reported successes of earlier workers and the more recent experiments of Perdrau. The experiments of the latter, as described, cannot be explained by the occasional existence of spontaneous encephalitis in his rabbits, nor by the assumption that a herpes virus fortuitously coexisted with that of lethargic encephalitis in his material, inasmuch as this material alone at first injection or in the unglycerolated state failed to infect. It is also possible to conceive that human beings may, by repeated skin infections, attain a not inconsiderable partial immunity to herpes virus, which would explain the nature of the clinical course (as in our partial immunity rabbits) as well as the innocuousness of direct injections of herpetic virus into man, as reported by Bastai and Busacca, and the finding of herpes virus in human beings not suffering from lethargic encephalitis. These suggestions are discussed in order to give this important problem the broadest possible consideration. For the time being, however, such reasoning cannot be taken as more than a logical possibility impressed upon us by our partial immunization experiments. All other experimental evidence obtained by direct inoculations with the limited material at our disposal tends to render identity of the two varieties of viruses unlikely.


Why fellowship? Peak professional bodies, peer recognition and credentialing in Australia.
Professions frequently distinguish themselves from trades or vocations, by creating professional peak bodies. Usually professional peak bodies offer two levels of subscription: membership and fellowship. Fellowship of a peak body is widely considered as offering status and prestige by recognising contribution made to one's profession. Fellowship therefore, often brings with it specific privileges of a symbolic or material nature, or both. To describe the criteria for eligibility for fellowship; the perceived benefits of fellowship to individuals from the organisations' viewpoint; the process of obtaining and maintaining fellowship across a range of professional peak bodies; and to apply the outcomes from the survey to nursing professional peak bodies. A telephone survey of 25 professional organisations representing a range of disciplines and industries in Australia was conducted. Each organisation was telephoned and the manager responsible for membership was invited to participate in a 10-min telephone survey. Of the 25 organisations approached, 24 agreed to participate (response rate 96%). For six (25%) of these participating organisations, membership was exclusively limited to nurses and/or midwives. Fellowship was offered in 22 organisations (92%). Significant professional experience (73%) was the most frequently cited criterion for fellowship. Professional leadership/contribution to the profession was listed by less than half (45%) organisations surveyed. Over half (55%) of those organisations that offered fellowship programs also were credentialing bodies. The main benefit of fellowship reported was the prospect of peer recognition and higher professional standing (77%). The majority of organisations reported offering fellowship status and all hadriteria for fellowship. The most frequently cited criteria were, however, based on longevity in the profession and continued financial membership rather than demonstrated leadership within the profession. There was variability in how fellowship was attained and maintained which may be linked to whether organisations have a credentialing role or not. A more in depth study of fellowship is warranted.


The role of religion in death attitudes: distinguishing between religious belief and style of processing religious contents.
Although it is widely assumed that religiosity plays an important role in individuals' attitudes about death, research to date has failed to reveal consistent associations between religiosity and death attitudes. Drawing from D. M. Wulff's (1991) multidimensional model of religiosity, the authors examined associations between religious attitudes as measured by the Post-Critical Belief Scale and death attitudes as assessed by the Death Attitude Profile--Revised. In total, 471 Dutch-speaking Belgian adults completed both questionnaires. Hierarchical multiple regression analyses were used to asses the unique contribution of the religious attitudes in the prediction of the death attitudes. First, results show that religious people are more likely to endorse an approach acceptance attitude toward death, indicating that religiosity as such is related to belief in an afterlife. Second, people holding a literal attitude toward religion report more death anxiety, indicating that the processing of religious contents is related to defensiveness toward death. Finally, the specific combination of the two dimensions seems important in the prediction of a neutral acceptance attitude. The relevance of our findings for future research on religiosity and death attitudes is discussed.


Porphyrin-lipid nanovesicles (Porphysomes) are effective photosensitizers for photodynamic therapy.
Porphysomes (PS) are liposome-like nanoparticles comprising pyropheophorbide-conjugated phospholipids that have demonstrated potential as multimodal theranostic agents for applications that include phototherapies, targeted drug delivery and in vivo fluorescence, photoacoustic, magnetic resonance or positron emission imaging. Previous therapeutic applications focused primarily on photothermal therapy (PTT) and suggested that PSs require target-triggered activation for use as photodynamic therapy (PDT) sensitizers. Here, athymic nude mice bearing subcutaneous A549 human lung tumors were randomized into treatment and control groups: PS-PDT at various doses, PS-only and no treatment negative controls, as well as positive controls using the clinical photosensitizer Photofrin. Animals were followed for 30 days post-treatment. PS-PDT at all doses demonstrated a significant tumor ablative effect, with the greatest effect seen with 10 mg/kg PS at a drug-light interval of 24 h. By comparison, negative controls (PS-only, Photofrin-only, and no treatment) showed uncontrolled tumor growth. PDT with Photofrin at 5 mg/kg and PS at 10 mg/kg demonstrated similar tumor growth suppression and complete tumor response rates (15 vs. 25%, p = 0.52). Hence, porphysome nanoparticles are an effective PDT agent and have the additional advantages of multimodal diagnostic and therapeutic applications arising from their intrinsic structure. Porphysomes may also be the first single all-organic agent capable of concurrent PDT and PTT.


Hypotheses for Near-Surface Exchange of Methane on Mars.
The Curiosity rover recently detected a background of 0.7 ppb and spikes of 7 ppb of methane on Mars. This in situ measurement reorients our understanding of the martian environment and its potential for life, as the current theories do not entail any geological source or sink of methane that varies sub-annually. In particular, the 10-fold elevation during the southern winter indicates episodic sources of methane that are yet to be discovered. Here we suggest a near-surface reservoir could explain this variability. Using the temperature and humidity measurements from the rover, we find that perchlorate salts in the regolith deliquesce to form liquid solutions, and deliquescence progresses to deeper subsurface in the season of the methane spikes. We therefore formulate the following three testable hypotheses. The first scenario is that the regolith in Gale Crater adsorbs methane when dry and releases this methane to the atmosphere upon deliquescence. The adsorption energy needs to be 36 kJ mol(-1) to explain the magnitude of the methane spikes, higher than existing laboratory measurements. The second scenario is that microorganisms convert organic matter in the soil to methane when they are in liquid solutions. This scenario does not require regolith adsorption but entails extant life on Mars. The third scenario is that deep subsurface aquifers produce the bursts of methane. Continued in situ measurements of methane and water, as well as laboratory studies of adsorption and deliquescence, will test these hypotheses and inform the existence of the near-surface reservoir and its exchange with the atmosphere. Mars-Methane-Astrobiology-Regolith. Astrobiology 16, 539-550.


The Stair Climb Power Test as an Efficacy Outcome in Randomized Trials of Function Promoting Therapies in Older Men.
Standardization of performance-based physical function measures that are reliable and responsive to intervention is necessary for efficacy trials of function promoting anabolic therapies (FPTs). Herein, we describe a standardized method of measuring stair climbing power (SCP) and evaluate its ability to assess improvements in physical function in response to an FPT (testosterone) compared to gait speed. We used a 12-step SCP test with and without carrying a load (loaded, LSCP or unloaded, USCP) in two testosterone trials in older men. SCP was determined from mass, total step-rise, and time of ascent measured with an electronic timing system. Associations between SCP and leg press performance (strength and power), testosterone levels, and gait speed were assessed. Test-retest reliability was evaluated using interclass correlation and Bland-Altman analyses. Baseline SCP was negatively associated with age and positively with leg strength and power and gait speed. Both tests of SCP were safe and showed excellent reliability (intra-class correlation 0.91-0.97 in both cohorts). Changes in testosterone concentrations were associated with changes in USCP and LSCP, but not gait speed in mobility-limited men. Changes in leg press performance were associated with SCP in both trials. Both USCP and LSCP are safe and have high test-retest reliability. Compared to gait speed, SCP is associated more robustly with leg press performance and is sensitive to testosterone therapy. The LSCP might be a more responsive outcome than gait speed to evaluate the efficacy of FPT in randomized trials.


Wettability of pristine and alkyl-functionalized graphane.
Graphane is a hydrogenated form of graphene with high bandgap and planar structure insensitive to a broad range of chemical substitutions. We describe an atomistic simulation approach to predict wetting properties of this new material. We determine the contact angle to be 73°. The lower hydrophobicity compared to graphene is explained by the increased planar density of carbon atoms while we demonstrate that the presence of partial charges on carbon and hydrogen atoms plays only a minor role. We further examine the effects of graphane functionalization by alkyl groups of increasing chain lengths. The gradual increase in contact angle with chain length offers a precise control of surface wettability. A saturated contact angle of 114° is reached in butylated form. We find the saturation of contact angle with respect to the length of the functional groups to coincide with the loss of water's ability to penetrate the n-alkyl molecular brush and interact with carbon atoms of the underlying lattice. Since no experimental data have yet become available, our modeling results provide the first estimate of the wettability of graphane. The results also show how its alkyl functionalization provides the basis for a variety of chemical modifications to tune hydrophilicity while preserving the planar geometry of the substrate.


A randomized trial of a multicomponent cessation strategy for emergency department smokers.
The objective was to determine the efficacy of an emergency department (ED)-based smoking cessation intervention. This study was a randomized trial conducted from January 2006 to September 2007 at an urban ED that treats 90,000 adults per year. Discharged adults who smoked at least 10 cigarettes per day were randomized to 1) usual care, receiving a smoking cessation brochure; or 2) enhanced care, receiving the brochure, a motivational interview (MI), nicotine patches, and a phone call at 3 days. Interventions were performed by a peer educator trained in tobacco treatment. Blinded follow-up was performed at 3 months. A total of 338 subjects were enrolled, mean (±SD) age was 40.2 (±12.0) years, 51.8% were female, and 56.5% were either self-pay or Medicaid. Demographic and clinical variables were comparable between groups. Enhanced and usual care arms showed similar cessation rates at 3 months (14.7% vs. 13.2%, respectively). The proportion of subjects making a quit attempt (69.2% vs. 66.5%) and decrease in daily cigarette use (five vs. one; all p > 0.05) were also similar. In logistic modeling, factors associated with quitting included any tobacco-related International Classification of Diseases, ninth revision (ICD-9), code for the ED visit (odds ratio [OR]= 3.42, 95% confidence interval [CI] = 1.61 to 7.26) or subject belief that the ED visit was tobacco-related (OR = 2.47, 95% CI = 1.17 to 5.21). Conversely, subjects who reported having a preexisting tobacco-related illness were less likely to quit (OR = 0.22, 95% CI = 0.10 to 0.50). The primary endpoint was negative, reflecting a higher-than-expected quit rate in the control group. Subjects whose ED visit was tobacco-related, based either on physician diagnosis or subject perception, were more than twice as likely to quit. These data suggest that even low-intensity screening and referral may prompt substantial numbers of ED smokers to quit or attempt to quit.


Genome-wide DNA methylation profiling by modified reduced representation bisulfite sequencing in Brassica rapa suggests that epigenetic modifications play a key role in polyploid genome evolution.
Brassica rapa includes some of the most important vegetables worldwide as well as oilseed crops. The complete annotated genome sequence confirmed its paleohexaploid origins and provides opportunities for exploring the detailed process of polyploid genome evolution. We generated a genome-wide DNA methylation profile for B. rapa using a modified reduced representation bisulfite sequencing (RRBS) method. This sampling represented 2.24% of all CG loci (2.5 × 10(5)), 2.16% CHG (2.7 × 10(5)), and 1.68% CHH loci (1.05 × 10(5)) (where H = A, T, or C). Our sampling of DNA methylation in B. rapa indicated that 52.4% of CG sites were present as (5m)CG, with 31.8% of CHG and 8.3% of CHH. It was found that genic regions of single copy genes had significantly higher methylation compared to those of two or three copy genes. Differences in degree of genic DNA methylation were observed in a hierarchical relationship corresponding to the relative age of the three ancestral subgenomes, primarily accounted by single-copy genes. RNA-seq analysis revealed that overall the level of transcription was negatively correlated with mean gene methylation content and depended on copy number or was associated with the different subgenomes. These results provide new insights into the role epigenetic variation plays in polyploid genome evolution, and suggest an alternative mechanism for duplicate gene loss.


Optics in Germany.
A survey is given of the current state of optics in the Federal Republic of Germany. Basic research is conducted at many universities and technical colleges, by government agencies and government sponsored research institutes, and in industry. All of these have made substantial contributions to modern optics. Applied research in conventional optics, directed toward the design and production of precision optical instrumentation, and toward the development of optical measurement techniques, continues to play an important role. Most of this work is carried out by Germany's precision engineering and optical industry-Education and training in opties is offered by many universities, technical colleges, and engineer's schools, and by industry.


Delivering optimal care to the person with asthma: what are the key components and what do we mean by patient education?
The new understanding of the pathogenesis of asthma has led to an increasing perception of the disease as a chronic condition, rather than one consisting of acute exacerbations. Health professionals need to help patients to accept this new understanding, and this involves a consideration of how best to deliver care and how best to enhance compliance. Good communication is essential, and structured patient education has now been shown to be cost effective. All health professionals need to think how education may best be incorporated into their care. Such care will involve the development of a partnership between health professional and the patient and their loved ones, and the acquisition by patients of skills in self-management. Several unknowns remain regarding who benefits more from group education as opposed to individual education, and some details of guided self-management. However, in the face of clear evidence of the benefits of patient education, these areas for further study should not delay the adoption of a strong commitment to education by all of those who care for those with asthma.


Association of renal function with vascular stiffness in older adults: the Rotterdam study.
arterial stiffening is a marker of vascular ageing and an independent risk factor for cardiovascular disease. A potential mechanism linking cardiovascular disease to chronic kidney disease might be the change in arterial elasticity. We aim to determine the association between renal function and arterial stiffness in older subjects. cross-sectional study. Rotterdam study, a population-based cohort study. we included 3,279 subjects from 1997 to 1999 with a mean age of 71.9 years. estimation of glomerular filtration rate (eGFR) was used to assess renal function. Aortic pulse wave velocity (PWV) and carotid distensibility coefficient were used as measures of arterial stiffness. each standard deviation increase in eGFR, adjusting for age and sex, was associated with 0.14 m/s lower PWV [95% confidence interval (CI): -0.23, -0.05]. Further adjustments for socio-demographic and cardiovascular risk factors did not change the association (β: -0.16 m/s; 95% CI: -0.26, -0.06). There was a linear association between mean values of PWV and quartiles of glomerular filtration rate (P for trend = 0.006). There was no association between decreased renal function and carotid distensibility. There was no statistical difference in the strength of the association between renal function and PWV in subgroups of participants with and without cardiovascular risk factors. in this large population-based study of elderly subjects, our findings suggest that renal impairment is associated with aortic stiffness. This association is independent of cardiovascular risk factors.


Effects of motivation on reward and attentional networks: an fMRI study.
Existing evidence suggests that reward and attentional networks function in concert and that activation in one system influences the other in a reciprocal fashion; however, the nature of these influences remains poorly understood. We therefore developed a three-component task to assess the interaction effects of reward anticipation and conflict resolution on the behavioral performance and the activation of brain reward and attentional systems. Sixteen healthy adult volunteers aged 21-45 years were scanned with functional magnetic resonance imaging (fMRI) while performing the task. A two-way repeated measures analysis of variance (ANOVA) with cue (reward vs. non-reward) and target (congruent vs. incongruent) as within-subjects factors was used to test for main and interaction effects. Neural responses to anticipation, conflict, and reward outcomes were tested. Behaviorally there were main effects of both reward cue and target congruency on reaction time. Neuroimaging results showed that reward anticipation and expected reward outcomes activated components of the attentional networks, including the inferior parietal and occipital cortices, whereas surprising non-rewards activated the frontoinsular cortex bilaterally and deactivated the ventral striatum. In turn, conflict activated a broad network associated with cognitive control and motor functions. Interaction effects showed decreased activity in the thalamus, anterior cingulated gyrus, and middle frontal gyrus bilaterally when difficult conflict trials (e.g., incongruent targets) were preceded by reward cues; in contrast, the ventral striatum and orbitofrontal cortex showed greater activation during congruent targets preceded by reward cues. These results suggest that reward anticipation is associated with lower activation in attentional networks, possibly due to increased processing efficiency, whereas more difficult, conflict trials are associated with lower activity in regions of the reward system, possibly because such trials are experienced as less rewarding.


Controlled follow-up study of physical and psychosocial functioning of adolescents with juvenile primary fibromyalgia syndrome.
Currently, there are no prospective studies exploring the prognosis of patients with juvenile primary FM syndrome (JPFS) or their physical, emotional and social outcomes as they enter the early adult years. The primary objective of this study was to assess long-term outcomes of a paediatric sample of clinically referred JPFS patients and their matched healthy controls. Participants were 48 youths (current mean age = 19 years) diagnosed with JPFS in childhood or adolescence and 43 healthy controls matched in age, gender and race. The average length of follow-up was 3.67 years (range 2-6 years). Participants completed online (web-based) self-report questionnaires about current pain and physical symptoms, health status, anxiety, depressive symptoms and current and past treatments. Results showed that 62.5% of participants in the JPFS group continued to experience widespread pain and 60.4% reported having all the cardinal features of FM syndrome (including widespread pain, poor sleep and fatigue) at follow-up. The JPFS group reported significantly lower scores on all measures of health status and physical functioning compared with healthy controls and significantly greater symptoms of anxiety and depression. The results of this controlled follow-up study demonstrate that symptoms of FM appear to be chronic in a majority of clinically referred JPFS patients and the associated physical and emotional impairment can also be persistent. Implications for treatment and the need for further prospective longitudinal studies are discussed.


Sphingomonas hankookensis sp. nov., isolated from wastewater.
A Gram-negative, non-motile, rod-shaped bacterial strain, ODN7(T), was isolated from a wastewater treatment plant in Korea, and its taxonomic position was investigated by use of a polyphasic taxonomic approach. Strain ODN7(T) grew optimally at around pH 7.5 and 30 degrees C and in the presence of 0-0.5 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain ODN7(T) fell within the cluster comprising species of the genus Sphingomonas, clustering with Sphingomonas panni C52(T), with which it shared highest 16S rRNA gene sequence similarity (98.9 %). The chemotaxonomic properties of strain ODN7(T) were consistent with those of the genus Sphingomonas. The predominant ubiquinone was Q-10, and the major fatty acids were C(18 : 1)omega7c, C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH, and C(16 : 0). Major polar lipids were sphingoglycolipid, phosphatidylcholine, phosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content of strain ODN7(T) was 67.2 mol%. Strain ODN7(T) exhibited levels of DNA-DNA relatedness of 15-32 % to the type strains of phylogenetically related Sphingomonas species and could be differentiated from these species based on differences in phenotypic characteristics. On the basis of the data presented, strain ODN7(T) is considered to represent a novel species of the genus Sphingomonas, for which the name Sphingomonas hankookensis sp. nov. is proposed. The type strain is ODN7(T) (=KCTC 22579(T)=CCUG 57509(T)).


Tumor-associated inflammation as a potential prognostic tool in BRCA1/2-associated breast cancer.
The prognosis of BRCA1/2-associated breast cancer partly depends on histologic characteristics. Most of these breast cancers, however, are poorly differentiated. BRCA1-associated cancers are mainly negative for estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. Consequently, the use of these histologic features for risk stratification in BRCA1/2 breast cancer is limited. We assessed the prognostic value of additional histologic features, including tumor-associated inflammation and tumor-associated stroma in BRCA1/2 breast cancer patients. From the Rotterdam Family Cancer Clinic database, we collected demographics, tumor characteristics, and follow-up data from female BRCA1/2 breast cancer patients. Tumor samples were centrally reviewed including histologic subtype, differentiation grade, tumor-associated inflammation density, amount of tumor-associated stroma, and intratumor necrosis. The impact of these factors on recurrence-free survival (RFS) was evaluated using univariate and multivariate Cox regression, adjusted for established prognostic features and year of diagnosis. We included 138 BRCA1 and 37 BRCA2 breast cancer patients. Median follow-up after diagnosis was 9.7 years. Independent prognostic factors for RFS were tumor size (hazard ratio [HR], 2.47 for >2 versus ≤2 cm; 95% confidence interval [95% CI], 1.10-5.57), tumor-associated inflammation (HR, 0.18 for moderate/marked versus absent/mild; 95% CI, 0.05-0.61), and intratumor necrosis (HR, 2.60 for presence versus absence; 95% CI, 1.12-6.05). Established prognostic factors as nodal status and differentiation grade were not significantly related to RFS. Subgroup analyses of 138 BRCA1 and 118 triple-negative breast cancer cases showed similar results. Tumor-associated inflammation density was the strongest predictor for RFS in this series of BRCA1/2 breast cancer patients. This provides a potential risk stratification tool that can easily be implemented in routine histologic examination.


The distribution of iron oxidation states in a constructed wetland as an indicator of its redox properties.
Wastewater-treatment processes taking place inside constructed wetlands are closely connected with chemical properties of these systems. The aeration of a wetland via the roots of the vegetation (and a subsequent formation of redox-potential gradients) strongly influences the wastewater treatment efficiency, and thus it represents one of the most important characteristics of the wetland. The concentration ratios of individual iron oxidation states (Fe(II) and Fe(III)) were determined as the indicator of the redox properties of the constructed wetland reed bed during this study. Interstitial water from the wetland was sampled eleven times throughout the year 2005. The spectrophotometric method using 1,10-phenanthroline was properly optimized (limits of detection and quantification, sensitivity, linear dynamic range, repeatability, and accuracy values were assessed) and applied for iron determination. Most of iron, ca. 98%, is reduced to the Fe(II) form in raw wastewater and water from the inflow zone of the constructed wetland, however, at the outflow and in the vegetation bed both iron oxidation states are usually detected. The presence of Fe(III) in the reed bed (ca. 10-30% for some samples) demonstrates the aeration of the wetland by the vegetation (Phragmites australis) resulting in a re-oxidation of Fe(II).


[Clinical characteristics of nontraumatic rhabdomyolysis in patients with liver cirrhosis].
Rhabdomyolysis is a serious and lethal condition that can be induced not only by traumatic causes but also by a variety of nontraumatic causes. However, there are few reports about rhabdomyolysis developed in patients with liver cirrhosis. We carried out this study to elucidate the clinical characteristics and courses of rhabdomyolysis in patients with liver cirrhosis. We analyzed 19 cases of nontraumatic rhabdomyolysis in patients with liver cirrhosis who had admitted at Korea University Ansan Hospital between October 2001 and September 2004. Alcohol (50%) was the main etiology of rhabdomyolysis in alcoholic liver cirrhosis patients, and the precipitating factors were not apparent (69.2%) in majority of nonalcoholic liver cirrhosis patients with rhabdomyolysis. Nonalcoholic liver cirrhosis patients had complaints of pain referable to the musculoskeletal system, but alcoholic liver cirrhosis patients had no typical complaints. Mortality of rhabdomyolysis in liver cirrhosis patients was high (42.1%), especially in decompensated liver cirrhosis patients (p=0.04). In nonalcoholic liver cirrhosis patients, the development of oliguria (p=0.007) and acute renal failure (p=0.049) in the course of rhabdomyolysis increased the mortality significantly. In cirrhosis patients, rhabdomyolysis showed a poor prognosis, especially in nonalcoholic liver cirrhosis with oliguria, acute renal failure, or decompensated liver cirrhosis. It is believed that a high clinical suspicion for the occurrence of rhabdomyolysis in liver cirrhosis patients can lead to quicker recognition and better patient care.


Newborn with neonatal form of molybdenum cofactor deficiency - the first patient in the Slovak Republic.
To present the case of a term newborn with rapid progression of signs of neurodegenerative disease. In a case of a term newborn with numerous dysmorphic features, with seizure activity from the 3rd day of life, hypertonia and serious changes on brain parenchyma were presented. Diagnosis of molybdenum cofactor deficiency was confirmed by the decreased level of uric acid, 31 μmol/l, in serum, increased excretion of thiosulfate and S-sulfocysteine in urine, taurine (1729.3 μmol/mmol crea; normal range 30-300 μmol/mmol crea) and xanthine (276.9 μmol/mmol crea; normal range < 25 μmol/mmol crea) in urine. Sulfite oxidase activity on skin fibroblasts in culture was not detectable. The patient died at the age of 28 days of life. Deficiency of molybdenum cofactor leads to accumulation of toxic metabolites (levels of sulfite), which causes disturbances of neurotransmitters even before delivery. Therapy is symptomatic, no effective therapy is available. Seizures are difficult to suppress. This case report is about the first patient in Slovakia.


Species-Specific Responses of Juvenile Rockfish to Elevated pCO2: From Behavior to Genomics.
In the California Current ecosystem, global climate change is predicted to trigger large-scale changes in ocean chemistry within this century. Ocean acidification-which occurs when increased levels of atmospheric CO2 dissolve into the ocean-is one of the biggest potential threats to marine life. In a coastal upwelling system, we compared the effects of chronic exposure to low pH (elevated pCO2) at four treatment levels (i.e., pCO2 = ambient [500], moderate [750], high [1900], and extreme [2800 μatm]) on behavior, physiology, and patterns of gene expression in white muscle tissue of juvenile rockfish (genus Sebastes), integrating responses from the transcriptome to the whole organism level. Experiments were conducted simultaneously on two closely related species that both inhabit kelp forests, yet differ in early life history traits, to compare high-CO2 tolerance among species. Our findings indicate that these congeners express different sensitivities to elevated CO2 levels. Copper rockfish (S. caurinus) exhibited changes in behavioral lateralization, reduced critical swimming speed, depressed aerobic scope, changes in metabolic enzyme activity, and increases in the expression of transcription factors and regulatory genes at high pCO2 exposure. Blue rockfish (S. mystinus), in contrast, showed no significant changes in behavior, swimming physiology, or aerobic capacity, but did exhibit significant changes in the expression of muscle structural genes as a function of pCO2, indicating acclimatization potential. The capacity of long-lived, late to mature, commercially important fish to acclimatize and adapt to changing ocean chemistry over the next 50-100 years is likely dependent on species-specific physiological tolerances.


A Screening Tool for the Direct Analysis of Marine and Freshwater Phycotoxins in Organic SPATT Extracts from the Chesapeake Bay.
Many detection methods for phycotoxins, bioactive compounds produced by harmful algae, focus on one compound or a class of related compounds. Multiple harmful algal species often co-occur in the environment, however, emphasizing the need to analyze for the presence of multiple groups of marine and freshwater phycotoxins in environmental samples, e.g., extracts from solid phase adsorption toxin tracking (SPATT). Two methods were developed to screen for 13 phycotoxins (microcystin-RR, -LR, -YR, azaspiracid-1, -2, karlotoxin 3, goniodomin A, brevetoxin-2, yessotoxin, pectenotoxin-2, dinophysistoxin-1, -2, and okadaic acid) in organic SPATT extracts using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) equipped with a trapping dimension (trap) and at-column dilution (ACD). The performance of each compound under 36 combinations of chromatographic conditions was characterized, and two final methods, acidic and basic, were selected based on peak shapes, signal intensities, resolution, and the separation in time of positive and negative MS ionization modes. Injection volumes of up to 1 mL were possible through trap/ACD technology, resulting in limits of detection between 0.001 and 0.05 µg/L across the analytes. Benefits highlighted in this study, beyond the improved detection limits and co-detection of multiple toxin groups, include the ability to inject samples of 100% organic solvent, ensuring analyte stability and streamlining workflow through the elimination of laborious sample preparation steps.


Amplitude- and Phase-Resolved Infrared Nanoimaging and Nanospectroscopy of Polaritons in a Liquid Environment.
Polaritons allow for strong light-matter coupling and for highly sensitive analysis of (bio)chemical substances and processes. Nanoimaging of the polaritons' evanescent fields is critically important for experimental mode identification and field confinement studies. Here we describe two setups for polariton nanoimaging and spectroscopy in liquid. We first demonstrate the mapping of localized plasmon polaritons in metal antennas with a transflection infrared scattering-type scanning near-field optical microscope (s-SNOM), where the tip acts as a near-field scattering probe. We then demonstrate a total internal reflection (TIR)-based setup, where the tip is both launching and probing ultraconfined polaritons in van der Waals materials (here phonon polaritons in hexagonal boron nitride flakes), laying the foundation for s-SNOM-based polariton interferometry in liquid. Our results promise manifold applications, for example, in situ studies of strong coupling between polaritons and molecular vibrations or chemical reactions at the bare or functionalized surfaces of polaritonic materials.


Choice of next-generation sequencing pipelines.
The next-generation sequencing (NGS) technologies are revolutionary tools which have made possible achieving remarkable advances in genetics since the beginning of the twenty-first century. Thanks to the possibility to produce large amount of sequence data, these tools are going to completely substitute other high-throughput technologies. Moreover, the large applications of NGS protocols are increasing the genetic decoding of biological systems through studies of genome anatomy and gene mapping, coupled to the transcriptome pictures. The application of NGS pipelines such as (1) de-novo genomic sequencing by mate-paired and whole-genome shotgun strategies; (2) specific gene sequencing on large bacterial communities; and (3) RNA-seq methods including whole transcriptome sequencing and Serial Analysis of Gene Expression (Sage-analysis) are fundamental in the genome-wide fields like metagenomics. Recently, the availability of these advanced protocols has allowed to overcome the usual sequencing technical issues related to the mapping specificity over standard shotgun library sequencing, the detection of large structural genomes variations and bridging sequencing gaps, as well as more precise gene annotation. In this chapter we will discuss how to manage a successful NGS pipeline from the planning of sequencing projects through the choice of the platforms up to the data analysis management.


Application of a New Statistical Model for Measurement Error to the Evaluation of Dietary Self-report Instruments.
Most statistical methods that adjust analyses for dietary measurement error treat an individual's usual intake as a fixed quantity. However, usual intake, if defined as average intake over a few months, varies over time. We describe a model that accounts for such variation and for the proximity of biomarker measurements to self-reports within the framework of a meta-analysis, and apply it to the analysis of data on energy, protein, potassium, and sodium from a set of five large validation studies of dietary self-report instruments using recovery biomarkers as reference instruments. We show that this time-varying usual intake model fits the data better than the fixed usual intake assumption. Using this model, we estimated attenuation factors and correlations with true longer-term usual intake for single and multiple 24-hour dietary recalls (24HRs) and food frequency questionnaires (FFQs) and compared them with those obtained under the "fixed" method. Compared with the fixed method, the estimates using the time-varying model showed slightly larger values of the attenuation factor and correlation coefficient for FFQs and smaller values for 24HRs. In some cases, the difference between the fixed method estimate and the new estimate for multiple 24HRs was substantial. With the new method, while four 24HRs had higher estimated correlations with truth than a single FFQ for absolute intakes of protein, potassium, and sodium, for densities the correlations were approximately equal. Accounting for the time element in dietary validation is potentially important, and points toward the need for longer-term validation studies.


Systematic Review of Factors Associated with the Utilization of Radical Cystectomy for Bladder Cancer.
Despite established guidelines for the treatment of muscle-invasive bladder cancer, it has been reported that radical cystectomy (RC) is markedly underused, especially among patients of advanced age and those with higher comorbidity burden and lower access to care. Understanding the interactions between patient, provider, and hospital factors may inform targeted interventions to optimize RC utilization. To systematically review the literature regarding factors associated with RC utilization. A systematic search was conducted using Ovid and Medline according to Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines to identify studies between 1970 and 2017 reporting on RC utilization. Prospective and retrospective studies were included. There are no published randomized control trials on RC utilization. Variations in study quality and design precluded a formal statistical meta-analysis. RC receipt significantly depended on patient, provider, and hospital factors. Patient factors associated with lower RC use included advanced age, African American and Hispanic race/ethnicity, higher comorbidity burden, unmarried marital status, higher tumor stage and grade, and lower socioeconomic status. Provider factors associated with underutilization included lower surgeon volume and a metropolitan location. Finally, hospital factors associated with lower RC use included low hospital volume, nonacademic affiliation, and hospital location in the Midwest. RC is reportedly underutilized. We found that age, race, marital status, socioeconomic factors, cancer severity, comorbidity burden, surgeon volume, and facility type and location significantly determined RC receipt. Improved understanding of the varying contributions of the risk factors according to patient, provider, and hospital determinants may assist in developing targeted interventions to improve RC utilization. In this review we explored the clinical evidence for factors predicting the utilization of radical cystectomy for muscle-invasive bladder cancer. Many factors related to the patient, provider, and hospital determine whether patients receive this guideline-recommended treatment. However, there remains a lack of understanding on characterization and targeted interventions according to these levels, which may improve use.


Epidemiology of Adult Diabetes Mellitus and its Correlates in Pars Cohort Study in Southern Iran.
The burden of diabetes mellitus (DM) is increasing worldwide, especially in countries such as Iran. Modifiable correlates of the DM may be different across regions. We aimed to estimate the population-based prevalence of DM and its correlates among adults in southern Iran. Baseline data of the Pars Cohort Study (PCS) was analyzed. Demographic and socio-economic characteristics, alcohol consumption, opium and tobacco abuse and cigarette smoking, laboratory findings, anthropometric measurements and perceived body shape at 15 and 30 years of age using pictogram were measured and analyzed. The age-standardized prevalence of DM was estimated. Robust Poisson regression was applied to estimate adjusted prevalence ratio (aPR) and 95% robust confidence intervals (CI). A P value <0.05 was considered statistically significant. Out of 9264 individuals, 919 (9.9%; 95% CI: 9.3-10.6) had DM. The prevalence of DM among the males and females was 7.6% (95% CI: 6.8-8.5) and 11.9% (95% CI: 10.9-12.9), respectively. Higher age, female gender, high socioeconomic status and using antihypertensive medication were associated with higher DM prevalence. Also, triglyceride level, physical inactivity, higher body mass index (BMI), pictogram score at 15, and its change from 15 to 30 years of age were significantly associated with DM. Higher prevalence of modifiable factors such as physical inactivity and obesity among DM patients in the study population highlights the necessity of more effective preventive interventions in such settings, especially in younger ages.


Recent trends in Pd-catalyzed remote functionalization of carbonyl compounds.
Recent advances in the palladium-catalyzed remote functionalization of carbonyl derivatives are highlighted in this review. The structure of the article is based on the three strategies that have emerged in recent years as the most viable tactics to construct C(sp(3))-C, C(sp(3))-N or C(sp(3))-O bonds at a distance of at least three carbon atoms from the carbonyl functionality. The specific aspects of each of these approaches are discussed in detail (reaction conditions, substrate scope and limitations). As the reading progresses, their complementarity should also appear clearly.


Menstrual cycle, pregnancy and oral contraceptive use alter attraction to apparent health in faces.
Previous studies demonstrating changes in women's face preferences have emphasized increased attraction to cues to possible indirect benefits (e.g. heritable immunity to infection) that coincides with periods of high fertility (e.g. the late follicular phase of the menstrual cycle). By contrast, here we show that when choosing between composite faces with raised or lowered apparent health, women's preferences for faces that are perceived as healthy are (i) stronger during the luteal phase of the menstrual cycle than during the late follicular, fertile phase, (ii) stronger in pregnant women than in non-pregnant women and (iii) stronger in women using oral contraceptives than in women with natural menstrual cycles. Change in preference for male faces was greater for short- than long-term relationships. These findings indicate raised progesterone level is associated with increased attraction to facial cues associated with possible direct benefits (e.g. low risk of infection) and suggest that women's face preferences are influenced by adaptations that compensate for weakened immune system responses during pregnancy and reduce the risk of infection disrupting foetal development.


Pre-transplant serum concentrations of anti-endothelial cell antibody in panel reactive antibody negative renal recipients and its impact on acute rejection.
Endothelial cell antigens are important targets in acute rejection (AR). Our goal was to measure the serum concentrations of pre-transplant anti-endothelial cell antibody (AECA) in panel reactive antibody (PRA) negative recipients and its impact on AR within 6 months following renal transplantation. We retrospectively examined pre-transplant sera from 392 patients using cellular enzyme linked immunosorbent assay (ELISA) with substrate from a permanent endothelial cell line EAhy926. Equal volumes of serum from 40 healthy volunteers were mixed and used as the negative control. The positive rate of AECA was 15.8%. There were no significant differences with respect to age, gender, original disease, dialysis history, immune suppressive regimen, cytomegalovirus (CMV) antigen positive rate, complement dependent cytotoxicity (CDC) level and soluble CD30 (sCD30) levels between the AECA positive group and AECA negative group. AR rate in the AECA positive group was higher than that in the AECA negative group (35.5% vs. 22.4%, p=0.023). The AECA positive patients had significantly higher rates of acute grade II T-cell mediated rejection (TMR) and acute antibody mediated rejection (AMR) compared with AECA negative patients. The concentrations of sCD30, and AECA were independent risk factors for AR within 6 months; the odds ratios were 7.005 and 2.469, respectively. Positive AECA was an independent risk factor for AR and appeared to correlate with relatively severe rejection subtypes.


An introduction to molecular imaging in radiation oncology: a report by the AAPM Working Group on Molecular Imaging in Radiation Oncology (WGMIR).
Molecular imaging is the direct or indirect noninvasive monitoring and recording of the spatial and temporal distribution of in vivo molecular, genetic, and/or cellular processes for biochemical, biological, diagnostic, or therapeutic applications. Molecular images that indicate the presence of malignancy can be acquired using optical, ultrasonic, radiologic, radionuclide, and magnetic resonance techniques. For the radiation oncology physicist in particular, these methods and their roles in molecular imaging of oncologic processes are reviewed with respect to their physical bases and imaging characteristics, including signal intensity, spatial scale, and spatial resolution. Relevant molecular terminology is defined as an educational assist. Current and future clinical applications in oncologic diagnosis and treatment are discussed. National initiatives for the development of basic science and clinical molecular imaging techniques and expertise are reviewed, illustrating research opportunities in as well as the importance of this growing field.


Development of an LC-MS/MS method to simultaneously monitor maitotoxins and selected ciguatoxins in algal cultures and P-CTX-1B in fish.
Ciguatera fish poisoning is a serious human health issue that is highly localized to tropical and sub-tropical coastal areas, affecting many of the indigenous island communities intrinsically linked to reef systems for sustenance and trade. It is caused by the consumption of reef fish contaminated with ciguatoxins and is reported as the most common cause of non-bacterial food poisoning. The causative toxins bioaccumulate up the food web, from small herbivorous fish that graze on microalgae of the genus Gambierdiscus into the higher trophic level omnivorous and carnivorous fish predating on them. The number of Gambierdiscus species being described is increasing rapidly and the role of other toxins produced by this microalgal genus in ciguatera intoxications, such as maitotoxin, remains unclear. Ciguatoxins and maitotoxin are among the most potent marine toxins known and there are currently no methods of analysis that can simultaneously monitor these toxins with a high degree of specificity. To meet this need a rapid and selective ultra-performance liquid chromatography tandem mass spectrometry method has been developed to rapidly screen Gambierdiscus cultures and environmental sample device extracts for ciguatoxins and maitotoxins. A fast sample preparation method has also been developed to allow sensitive quantification of the potent ciguatoxin fish metabolite P-CTX-1B from fish extracts, and this method has been subjected to a small validation study. Novel aspects of this approach include the use of alkaline mobile phase for chromatographic separation and specific monitoring of the various toxins. This method has good potential to help evaluate ciguatera risk associated with Gambierdiscus and related microalgal species, and to help promote method development activities for this important and analytically challenging toxin class.


Maternal hypoxic ventilatory response, ventilation, and infant birth weight at 4,300 m.
To test the hypothesis that increased hypoxic ventilatory responsiveness (HVR) raised maternal ventilation and arterial oxygenation during high-altitude pregnancy and related to the birth weight of the offspring, we studied 21 residents of Cerro de Pasco, Peru (4,300 m), while eight of them were 36 +/- 0 wk pregnant and 15 of them 13 +/- 0 wk postpartum. HVR was low in the nonpregnant women (mean +/- SE shape parameter A = 23 +/- 8) but increased nearly fourfold with pregnancy (A = 87 +/- 17). The increase in HVR appeared to account for the 25% rise in resting ventilation with pregnancy (delta VE observed = 2.4 +/- 0.7 l/min BTPS vs. delta VE predicted from delta HVR = 2.6 +/- 1.7 l/min BTPS, P = NS). Hyperoxia decreased ventilation in the pregnant women (P less than 0.01) to levels similar to those measured when nonpregnant. The increased ventilation of pregnancy raised arterial O2 saturation (SaO2) from 83 +/- 1 to 87 +/- 0%, and SaO2 was correlated positively with HVR in the pregnant women. The rise in SaO2 compensated for a 0.9 g/100 ml decrease in hemoglobin concentration to preserve arterial O2 content at levels present when nonpregnant. Cardiac output in the 36th wk of pregnancy did not differ significantly from values measured postpartum. The increase in HVR correlated positively with infant birth weight. An increase in HVR may be an important contributor to increased maternal ventilation with pregnancy and infant birth weight at high altitude.


Prediction of Incident Major Osteoporotic and Hip Fractures by Trabecular Bone Score (TBS) and Prevalent Radiographic Vertebral Fracture in Older Men.
Trabecular bone score (TBS) has been shown to predict major osteoporotic (clinical vertebral, hip, humerus, and wrist) and hip fractures in postmenopausal women and older men, but the association of TBS with these incident fractures in men independent of prevalent radiographic vertebral fracture is unknown. TBS was estimated on anteroposterior (AP) spine dual-energy X-ray absorptiometry (DXA) scans obtained at the baseline visit for 5979 men aged ≥65 years enrolled in the Osteoporotic Fractures in Men (MrOS) Study and its association with incident major osteoporotic and hip fractures estimated with proportional hazards models. Model discrimination was tested with Harrell's C-statistic and with a categorical net reclassification improvement index, using 10-year risk cutpoints of 20% for major osteoporotic and 3% for hip fractures. For each standard deviation decrease in TBS, there were hazard ratios of 1.27 (95% confidence interval [CI] 1.17 to 1.39) for major osteoporotic fracture, and 1.20 (95% CI 1.05 to 1.39) for hip fracture, adjusted for FRAX with bone mineral density (BMD) 10-year fracture risks and prevalent radiographic vertebral fracture. In the same model, those with prevalent radiographic vertebral fracture compared with those without prevalent radiographic vertebral fracture had hazard ratios of 1.92 (95% CI 1.49 to 2.48) for major osteoporotic fracture and 1.86 (95% CI 1.26 to 2.74) for hip fracture. There were improvements of 3.3%, 5.2%, and 6.2%, respectively, of classification of major osteoporotic fracture cases when TBS, prevalent radiographic vertebral fracture status, or both were added to FRAX with BMD and age, with minimal loss of correct classification of non-cases. Neither TBS nor prevalent radiographic vertebral fracture improved discrimination of hip fracture cases or non-cases. In conclusion, TBS and prevalent radiographic vertebral fracture are associated with incident major osteoporotic fractures in older men independent of each other and FRAX 10-year fracture risks, and these data support their use in conjunction with FRAX for fracture risk assessment in older men.


A computational model of prefrontal control in free recall: strategic memory use in the California Verbal Learning Task.
Several decades of research into the function of the frontal lobes in brain-damaged patients, and more recently in intact individuals using function brain imaging, has delineated the complex executive functions of the frontal cortex. And yet, the mechanisms by which the brain achieves these functions remain poorly understood. Here, we present a computational model of the role of the prefrontal cortex (PFC) in controlled memory use that may help to shed light on the mechanisms underlying one aspect of frontal control: the development and deployment of recall strategies. The model accounts for interactions between the PFC and medial temporal lobe in strategic memory use. The PFC self-organizes its own mnemonic codes using internally derived performance measures. These mnemonic codes serve as retrieval cues by biasing retrieval in the medial temporal lobe memory system. We present data from three simulation experiments that demonstrate strategic encoding and retrieval in the free recall of categorized lists of words. Experiment 1 compares the performance of the model with two control networks to evaluate the contribution of various components of the model. Experiment 2 compares the performance of normal and frontally lesioned models to data from several studies using frontally intact and frontally lesioned individuals, as well as normal, healthy individuals under conditions of divided attention. Experiment 3 compares the model's performance on the recall of blocked and unblocked categorized lists of words to data from Stuss et al. (1994) for individuals with control and frontal lobe lesions. Overall, our model captures a number of aspects of human performance on free recall tasks: an increase in total words recalled and in semantic clustering scores across trials, superiority on blocked lists of related items compared to unblocked lists of related items, and similar patterns of performance across trials in the normal and frontally lesioned models, with poorer overall performance of the lesioned models on all measures. The model also has a number of shortcomings, in light of which we suggest extensions to the model that would enable more sophisticated forms of strategic control.


Resting energy expenditure measured longitudinally following hip fracture compared to predictive equations: is an injury adjustment required?
The present study measuring resting energy expenditure (REE; kJ/d) longitudinally using indirect calorimetry in six elderly women aged > or =70 years following surgery for hip fracture, describes changes over time (days 10, 42 and 84 post-injury) and compares measured values to those calculated from routinely applied predictive equations. REE was compared to REE predicted using the Harris Benedict and Schofield equations, with and without accounting for the theoretical increase in energy expenditure of 35 % secondary to physiological stress of injury and surgery. Mean (95 % CI) measured REE (kJ/d) was 4704 (4354, 5054), 4090 (3719, 4461) and 4145 (3908, 4382) for days 10, 42 and 84, respectively. A time effect was observed for measured REE, P=0.003. Without adjusting for stress the mean difference and 95 % limits of agreement for measured and predicted REE (kJ/kg per d) for the Harris Benedict equation were 1 (-9, 12), 10 (2, 18) and 9 (1, 17) for days 10, 42 and 84, respectively. The mean difference and 95 % limits of agreement for measured and predicted REE (kJ/kg per d) for the Schofield equation without adjusting for stress were 8 (-3, 19), 16 (6, 26) and 16 (10, 22) for days 10, 42 and 84, respectively. After adjusting for stress, REE predicted from the Harris Benedict or Schofield equations overestimated measured REE by between 38 and 69 %. Energy expenditure following fracture is poorly understood. Our data suggest REE was relatively elevated early in recovery but declined during the first 6 weeks. Using the Harris Benedict or Schofield equations adjusted for stress may lead to overestimation of REE in the clinical setting. Further work is required to evaluate total energy expenditure before recommendations can be made to alter current practice for calculating theoretical total energy requirements of hip fracture patients.


Fitness correlates of song repertoire size in free-living song sparrows (Melospiza melodia).
Models of sexual selection propose that exaggerated secondary sexual ornaments indicate a male's own fitness and the fitness of his offspring. These hypotheses have rarely been thoroughly tested in free-living individuals because overall fitness, as opposed to fitness components, is difficult to measure. We used 20 years of data from song sparrows (Melospiza melodia) inhabiting Mandarte Island, British Columbia, Canada, to test whether a male's song repertoire size, a secondary sexual trait, predicted overall measures of male or offspring fitness. Males with larger song repertoires contributed more independent and recruited offspring, and independent and recruited grandoffspring, to Mandarte's population. This was because these males lived longer and reared a greater proportion of hatched chicks to independence from parental care, not because females mated to males with larger repertoires laid or hatched more eggs. Furthermore, independent offspring of males with larger repertoires were more likely to recruit and then to leave more grandoffspring than were offspring of males with small repertoires. Although we cannot distinguish whether observed fitness variation reflected genetic or environmental effects on males or their offspring, these data suggest that female song sparrows would gain immediate and intergenerational fitness benefits by pairing with males with large song repertoires.


Personality characteristics ascribed to young women who drink alcohol.
This study employs an implicit personality rating methodology from marketing research (the "shopping list" method) to investigate young adults' perceptions of young women who drink alcoholic beverages. Whereas there were some differences in perceptions of young women drinkers across the alcohol brands, the greatest differential in the personality ratings was caused by the mere presence or absence of alcohol. Young women who drink alcohol were described as being more "interesting" and "self-assured" than young women who do not drink. These positive traits were reliably ascribed to young women drinkers by both young adult men and young adult women and more strongly by those who themselves drink. Although it is possible that young women drinkers really are differentially interesting and self-assured, a more likely contributor to these perceptions is advertising aimed at the female drinker.


Wnt4 expression in the differentiating gonad of the frog Rana rugosa.
Wnt4, a member of the Wnt family, is known to influence the sex-determination cascade. In mice having a targeted deletion of Wnt4, masculinization occurs in XX pups. Therefore, in addition to Sry, Wnt4 is also involved in sex determination in mice. In humans, a chromosomal duplication of the WNT4 causes feminization of XY-individuals. Thus, for better understanding of the mechanism of sex determination in vertebrates, it is necessary to examine the expression of Wnt4 at early gonadal development stages in non-mammalians. We first isolated the Wnt4 cDNA from the tetsis of the frog Rana rugosa. R. rugosa Wnt4 had a high similarity (>86%) at the amino acid level with zebra fish, chicken, mouse, and human Wnt4s. We next employed RT-PCR analysis to examine whether Wnt4 was expressed in a sexually dimorphic fashion at early stages of gonadal development in R. rugosa. Wnt4 was transcribed first in the embryos at the late gastrula stage, and its expression was maintained until the indifferent gonad differentiated into a testis or an ovary. Wnt4 expression in the differentiating gonad appeared in a non-sexually dimorphic pattern. The results, taken together, suggest that Wnt4 is highly conserved through evolution, and that its expression in the indifferent gonad takes place with no sexual dimorphism. Thus, Wnt4 is not a key factor to initiate the development of a testis or an ovary from the indifferent gonads in R. rugosa. However, this gene probably forms part of a gonadal-development pathway in this species.


Structures of mismatch replication errors observed in a DNA polymerase.
Accurate DNA replication is essential for genomic stability. One mechanism by which high-fidelity DNA polymerases maintain replication accuracy involves stalling of the polymerase in response to covalent incorporation of mismatched base pairs, thereby favoring subsequent mismatch excision. Some polymerases retain a "short-term memory" of replication errors, responding to mismatches up to four base pairs in from the primer terminus. Here we a present a structural characterization of all 12 possible mismatches captured at the growing primer terminus in the active site of a polymerase. Our observations suggest four mechanisms that lead to mismatch-induced stalling of the polymerase. Furthermore, we have observed the effects of extending a mismatch up to six base pairs from the primer terminus and find that long-range distortions in the DNA transmit the presence of the mismatch back to the enzyme active site, suggesting the structural basis for the short-term memory of replication errors.


Clinical efficacy of percutaneous renal revascularization with stent placement in hypertension among patients with atherosclerotic renovascular diseases.
The aim was to assess the effect of renal angioplasty with stent on systolic, diastolic, and mean arterial blood pressure (MAP) in awake and sleep time with ambulatory blood pressure (ABP) monitoring (Holter monitoring). Patients with angiographically proven atherosclerotic renal artery stenosis (RAS) were referred to the Angiography Department of Imam Hospital for intervention during a 1-year period from June 2008 to December 2009. Primary stent placement was attempted by a single operator in 27 severe RAS cases although 1 case was omitted from the study because of technical failure. Pre- and postprocedure creatinine levels, ejection fraction (EF), history of diabetes mellitus (DM), and ABP were obtained. Twenty-six (17 men, 9 women; average age, 62.6 years; age range, 90-21 years) consecutive patients participated in the study. All patients had severe hypertension resistant to multiple medications; 10 patients had impaired renal function (serum creatinine level greater than 130 µmol/L). A total of 3 (11.5%) patients had congestive heart failure, and 10 (37.7%) were diabetic. Hypertension was cured in 1 (4%) patient, had improved in 23 (88.4%) patients, and had failed to respond to treatment in 2 (7.6%). Serum creatinine decreased significantly from 1.46 ± 0.89 to 1.35 ± 0.61 mg/dL (P<0.05). Percutaneous transluminal angioplasty for atheromatous RAS rarely cures hypertension, but improved blood pressure control is often achieved.


Regulation and role of REST and REST4 variants in modulation of gene expression in in vivo and in vitro in epilepsy models.
Repressor element-1 silencing transcription factor (REST) is a candidate modulator of gene expression during status epilepticus in the rodent. In such models, full-length REST and the truncated REST4 variant are induced and can potentially direct differential gene expression patterns. We have addressed the regulation of these REST variants in rodent hippocampal seizure models and correlated this with expression of the proconvulsant, substance P encoding, PPT-A gene. REST and REST4 were differentially regulated following kainic acid stimulus both in in vitro and in vivo models. REST4 was more tightly regulated than REST in both models and its transient expression correlated with that of the differential regulation of PPT-A. Consistent with this, overexpression of a truncated REST protein (HZ4, lacking the C-terminal repression domain) increased expression of the endogenous PPT-A gene. Similarly the proximal PPT-A promoter reporter gene construct was differentially regulated by the distinct REST isoforms in hippocampal cells with HZ4 being the major inducer of increased reporter expression. Furthermore, REST and REST4 proteins were differentially expressed and compartmentalized within rat hippocampal cells in vitro following noxious stimuli. This differential localization of the REST isoforms was confirmed in the CA1 region following perforant path and kainic acid induction of status epilepticus in vivo. We propose that the interplay between REST and REST4 alter the expression of proconvulsant genes, as exemplified by the PPT-A gene, and may therefore regulate the progression of epileptogenesis.


Parasites of sheep herding dogs in central Germany.
This paper reports on endoparasite infections diagnosed in 2012 by standard coproscopical techniques and coproantigen Giardia ELISA in 165 dogs used for sheep herding in 36 farms in central Germany. The overall prevalence of dogs with evidence of endoparasite infections was 27.3% (95% CI 20.6-34.7). The most frequently identified faecal forms were those of ascarids (Toxocara, 6.7%; Toxascaris 3.6%), hookworms (5.5%) and taeniid cestodes (4.2%), followed by those of Trichuris whipworms (3.0%), Capillaria aerophila (1.8%), Angiostrongylus and Crenosoma lungworms (1.2% each) and Cystoisospora canis coccidians (0.6%). Molecular identification demonstrated the seven dogs shedding taeniid eggs positive for Taenia (T.) species tapeworms (five, T. hydatigena; one, T. ovis; one Taenia sp.). Screening of the faeces with the coproantigen ELISA revealed Giardia specific antigen in 5.5% of the samples. The majority of the dogs had evidence of single endoparasite infections (22.4%) while evidence for infection with two or three parasites concurrently was found in six (3.6%) and two (1.2%) of the dogs, respectively. Dogs ≤ 1 year (n = 19) were parasitized more frequently (p < 0.05) with overall gastrointestinal parasites (63.2% vs. 20.5%), ascarids (36.8% vs. 6.8%) and Giardia spp. (21.1% vs. 3.4%) than older dogs (n = 146). Dogs which had been wormed within six months of examination tested less frequently positive for gastrointestinal helminths compared to dogs not wormed (11.1% vs. 25.0%; p = 0.0567). In addition, ear swabs taken from 43 sheep dogs in 2012 were examined, and Otodectes cynotis mites were extracted from one dog. Identification of ectoparasites collected by full body search and combing from 113 sheep dogs in the years 2011 to 2013 revealed infestation of fleas and ticks (each up to five specimens per dog) on 13 and 108 dogs, respectively, with nine dogs carrying both fleas and ticks. Archaeopsylla erinacei, Ctenocephalides (C) canis, C. felis and Pulex irritans fleas were identified from one, three, two and eleven dogs, respectively. Dermacentor reticulatus, Ixodes (I.) hexagonus and I. ricinus ticks were isolated from 16, eight and 95 dogs, respectively.