Carboxydotrophy potential of uncultivated Hydrothermarchaeota from the subseafloor crustal biosphere.
The exploration of Earth's terrestrial subsurface biosphere has led to the discovery of several new archaeal lineages of evolutionary significance. Similarly, the deep subseafloor crustal biosphere also harbors many unique, uncultured archaeal taxa, including those belonging to Candidatus Hydrothermarchaeota, formerly known as Marine Benthic Group-E. Recently, Hydrothermarchaeota was identified as an abundant lineage of Juan de Fuca Ridge flank crustal fluids, suggesting its adaptation to this extreme environment. Through the investigation of single-cell and metagenome-assembled genomes, we provide insight into the lineage's evolutionary history and metabolic potential. Phylogenomic analysis reveals the Hydrothermarchaeota to be an early-branching archaeal phylum, branching between the superphylum DPANN, Euryarchaeota, and Asgard lineages. Hydrothermarchaeota genomes suggest a potential for dissimilative and assimilative carbon monoxide oxidation (carboxydotrophy), as well as sulfate and nitrate reduction. There is also a prevalence of chemotaxis and motility genes, indicating adaptive strategies for this nutrient-limited fluid-rock environment. These findings provide the first genomic interpretations of the Hydrothermarchaeota phylum and highlight the anoxic, hot, deep marine crustal biosphere as an important habitat for understanding the evolution of early life.


The Hans Eysenck affair: Time to correct the scientific record.
The Journal of Health Psychology publishes here Dr Anthony Pelosi's analysis of questionable science by one of the world's best-known psychologists, the late Professor Hans J Eysenck. The provenance of a huge body of data produced by Eysenck and Ronald Grossarth-Maticek is highly controversial. In Open letters to King's College London and the British Psychological Society, this editor is requesting a thorough investigation of the facts together with retraction or correction of 61 publications. Academic institutions have a conflict of interest concerning allegations of misconduct, which is why I believe that the only way forward is to have a National Research Integrity Ombudsperson to investigate allegations.


Differences in the predictors of reading comprehension in first graders from low socio-economic status families with either good or poor decoding skills.
Based on the assumption that good decoding skills constitute a bootstrapping mechanism for reading comprehension, the present study investigated the relative contribution of the former skill to the latter compared to that of three other predictors of reading comprehension (listening comprehension, vocabulary and phonemic awareness) in 392 French-speaking first graders from low SES families. This large sample was split into three groups according to their level of decoding skills assessed by pseudoword reading. Using a cutoff of 1 SD above or below the mean of the entire population, there were 63 good decoders, 267 average decoders and 62 poor decoders. 58% of the variance in reading comprehension was explained by our four predictors, with decoding skills proving to be the best predictor (12.1%, 7.3% for listening comprehension, 4.6% for vocabulary and 3.3% for phonemic awareness). Interaction between group versus decoding skills, listening comprehension and phonemic awareness accounted for significant additional variance (3.6%, 1.1% and 1.0%, respectively). The effects on reading comprehension of decoding skills and phonemic awareness were higher in poor and average decoders than in good decoders whereas listening comprehension accounted for more variance in good and average decoders than in poor decoders. Furthermore, the percentage of children with impaired reading comprehension skills was higher in the group of poor decoders (55%) than in the two other groups (average decoders: 7%; good decoders: 0%) and only 6 children (1.5%) had impaired reading comprehension skills with unimpaired decoding skills, listening comprehension or vocabulary. These results challenge the outcomes of studies on "poor comprehenders" by showing that, at least in first grade, poor reading comprehension is strongly linked to the level of decoding skills.


Innovative methods to assess upper limb strength and function in non-ambulant Duchenne patients.
Upper limb assessment in non-ambulant patients remains a challenge. We have designed new tools to precisely assess pinch (MyoPinch), grip (MyoGrip), wrist flexion and extension (MyoWrist) strength. We have also designed a new tool to assess the ability of patients to produce repetitive flexion/extension movements of wrist and fingers (MoviPlate). We have assessed the feasibility and reliability of these new tools in 30 non-ambulant patients with Duchenne muscular dystrophy and in 30 age-matched male controls. Existing measures, such as Motor Function Measure, Tapping, and the Brooke Upper Extremity Functional Rating Scale were also performed. Results demonstrated that assessments were feasible in nearly all upper limbs tested for MyoGrip, MyoPinch and MoviPlate. The reliability of all tests, including MyoWrist which was not feasible in the patients presenting with contractures, was excellent in patients as in controls. Motor capacities decrease with the number of months spent in the wheelchair. The scores in the tests were partially correlated with each other, and with clinical measures such as vital capacity, Motor Function Measure, functional hand scale and Brooke score. This study validates a panel of upper limb muscle strength and function measures for Duchenne Muscular Dystrophy which can be applied from controls to extremely weak patients.


Hospitalization rates and survival associated with COPD: a nationwide Danish cohort study.
We examined rates of first hospitalization for COPD, rates of 5-year mortality among patients hospitalized for COPD, and comparisons of mortality between COPD patients and a matched cohort free of COPD. We computed standardized rates of first COPD hospitalization. Using Cox regression, we compared 180- and 181-day to 5-year mortality among COPD patients with the comparison cohort. We used medical databases in Denmark (population 5.4 million) from 1997 to 2006. We included patients 40 years or older with first hospitalization for COPD (64,499) and an age- and gender-matched comparison cohort of persons without COPD hospitalization (322,495). We examined the incidence of COPD hospitalization and risks and rates of mortality in the 5 years after hospitalization. Standardized rates of first hospitalization for COPD declined from 276 per 100,000 person-years in 1999 to 231 per 100,000 person-years in 2006. Within 180 days of hospitalization, 16% of COPD patients and 2.4% of persons in the matched cohort died (adjusted hazard ratio = 7.00, 95% CI = 6.79-7.22). Between 181 days and 5 years, 46% of COPD patients who survived the first 180 days and 19% of persons in the comparison cohort died (adjusted hazard ratio = 2.91, 95% CI = 2.86-2.95). COPD and comorbid diseases interacted to increase mortality in the first 180 days, but not thereafter. COPD continues to be a major public health problem causing substantial mortality in the 5 years after hospitalization, particularly in the first 180 days and in patients with comorbid diseases.


Nodal specifies embryonic visceral endoderm and sustains pluripotent cells in the epiblast before overt axial patterning.
Anteroposterior (AP) polarity in the mammalian embryo is specified during gastrulation when naive progenitor cells in the primitive ectoderm are recruited into the primitive streak to form mesoderm and endoderm. At the opposite pole, this process is inhibited by signals previously induced in distal visceral endoderm (DVE). Both DVE and primitive streak formation, and hence positioning of the AP axis, rely on the TGFbeta family member Nodal and its proprotein convertases Furin and Pace4. Here, we show that Nodal and Furin are initially co-expressed in the primitive endoderm together with a subset of DVE markers such as Lefty1 and Hex. However, with the appearance of extra-embryonic ectoderm (ExE), DVE formation is transiently inhibited. During this stage, Nodal activity is essential to specify embryonic VE and restrict the expression of Furin to the extra-embryonic region. Activation of Nodal is also necessary to maintain determinants of pluripotency such as Oct4, Nanog and Foxd3 during implantation, and to stimulate elongation of the egg cylinder, before inducing DVE and germ layer formation. We conclude that Nodal is already activated in primitive endoderm, but induces a functional DVE only after promoting the expansion of embryonic VE and pluripotent progenitor cells in the epiblast.


Deep-sea record of impact apparently unrelated to mass extinction in the Late Triassic.
The 34-million-year (My) interval of the Late Triassic is marked by the formation of several large impact structures on Earth. Late Triassic impact events have been considered a factor in biotic extinction events in the Late Triassic (e.g., end-Triassic extinction event), but this scenario remains controversial because of a lack of stratigraphic records of ejecta deposits. Here, we report evidence for an impact event (platinum group elements anomaly with nickel-rich magnetite and microspherules) from the middle Norian (Upper Triassic) deep-sea sediment in Japan. This includes anomalously high abundances of iridium, up to 41.5 parts per billion (ppb), in the ejecta deposit, which suggests that the iridium-enriched ejecta layers of the Late Triassic may be found on a global scale. The ejecta deposit is constrained by microfossils that suggest correlation with the 215.5-Mya, 100-km-wide Manicouagan impact crater in Canada. Our analysis of radiolarians shows no evidence of a mass extinction event across the impact event horizon, and no contemporaneous faunal turnover is seen in other marine planktons. However, such an event has been reported among marine faunas and terrestrial tetrapods and floras in North America. We, therefore, suggest that the Manicouagan impact triggered the extinction of terrestrial and marine organisms near the impact site but not within the pelagic marine realm.


Nanoporous Ge electrode as a template for nano-sized ( <5 nm) Au aggregates.
In this paper we present the extremely peculiar electrical properties of nanoporous Ge. A full and accurate electrical characterization showed an unexpected and extremely high concentration of positive carriers. Electrochemical analyses showed that nanoporous Ge has improved charge transfer properties with respect to bulk Ge. The electrode behavior, together with the large surface-to-volume ratio, make nanoporous Ge an efficient nanostructured template for the realization of other porous materials by electrodeposition. The pores were efficiently decorated by Au nanoparticles of diameter as low as 1-5 nm, prepared by electrochemical deposition. These new results demonstrate the potential and efficient use of nanoporous Ge as a nanostructured template for nano-sized Au aggregates, opening the way for the realization of innovative sensor devices.


Production of highly concentrated and hyperpolarized metabolites within seconds in high and low magnetic fields.
Hyperpolarized metabolites are very attractive contrast agents for in vivo magnetic resonance imaging studies enabling early diagnosis of cancer, for example. Real-time production of concentrated solutions of metabolites is a desired goal that will enable new applications such as the continuous investigation of metabolic changes. To this end, we are introducing two NMR experiments that allow us to deliver high levels of polarization at high concentrations (50 mM) of an acetate precursor (55% 13C polarization) and acetate (17% 13C polarization) utilizing 83% para-state enriched hydrogen within seconds at high magnetic field (7 T). Furthermore, we have translated these experiments to a portable low-field spectrometer with a permanent magnet operating at 1 T. The presented developments pave the way for a rapid and affordable production of hyperpolarized metabolites that can be implemented in e.g. metabolomics labs and for medical diagnosis.


p53: new roles in metabolism.
Virtually all cancers show metabolic changes that result in upregulation of glycolysis and glucose consumption. Although discovered in the 1920s, how this glycolytic switch happens, and whether it is a cause or a consequence of the malignant process, has remained a matter of debate. The p53 tumor suppressor gene, discovered some 30 years ago, has a key role in preventing cancer development. Recent discoveries revealing new functions for p53 in the regulation of glucose metabolism and oxidative stress have brought together these two venerable fields of cancer biology. These activities of p53 appear to be key in tumor suppression, and shed some light on the pathways that underlie the metabolic changes in cancer cells.


The magnitude of global marine species diversity.
The question of how many marine species exist is important because it provides a metric for how much we do and do not know about life in the oceans. We have compiled the first register of the marine species of the world and used this baseline to estimate how many more species, partitioned among all major eukaryotic groups, may be discovered. There are ∼226,000 eukaryotic marine species described. More species were described in the past decade (∼20,000) than in any previous one. The number of authors describing new species has been increasing at a faster rate than the number of new species described in the past six decades. We report that there are ∼170,000 synonyms, that 58,000-72,000 species are collected but not yet described, and that 482,000-741,000 more species have yet to be sampled. Molecular methods may add tens of thousands of cryptic species. Thus, there may be 0.7-1.0 million marine species. Past rates of description of new species indicate there may be 0.5 ± 0.2 million marine species. On average 37% (median 31%) of species in over 100 recent field studies around the world might be new to science. Currently, between one-third and two-thirds of marine species may be undescribed, and previous estimates of there being well over one million marine species appear highly unlikely. More species than ever before are being described annually by an increasing number of authors. If the current trend continues, most species will be discovered this century.


Patient activation in Europe: an international comparison of psychometric properties and patients' scores on the short form Patient Activation Measure (PAM-13).
To allow better assessment of patients' individual competencies for self-management, the Patient Activation Measure (PAM) has been developed in the USA. Because the American studies have shown the PAM to be a valuable tool, several European countries have translated the instrument into their native languages (Danish, Dutch, German, Norwegian). The aim was to compare the psychometric properties in studies from the different countries and establish whether the scores on the PAM vary between the studies. Data from the four separate studies were subjected to the same data cleaning procedures and statistical analyses. The psychometric properties of the instruments were established with measures of data quality and scale structure. The mean patient activation score and distribution across four predefined activation levels were described and the differences between the four studies were tested with ANOVA (unadjusted and adjusted) followed by a post-hoc Tukey HSD test and the Pearson chi-squared test respectively. The total N of the four studies was 5184. The percentage of missing values was low in all datasets, confirming the good quality of the datasets. Factor analyses revealed moderate to strong factor loadings on the first factor in all datasets. Cronbach's α was high for all version, ranging from .80 (German) to .88 (Dutch). Item-rest correlations varied between .32 and .66, indicating a moderate to strong correlation of the individual items to the sum scale. Both the mean PAM score and the distribution across activation levels differed between the four datasets. After adjustment of the PAM score, patients in Norway in particular had a higher patient activation level. The European translations of PAM-13 (into Danish, Dutch, German and Norwegian) resulted in four instruments with good psychometric capabilities for measuring patient activation. The mean PAM score and the distribution across activation levels differed between the four datasets.


Two new monoclonal antibodies (LN-1, LN-2) reactive in B5 formalin-fixed, paraffin-embedded tissues with follicular center and mantle zone human B lymphocytes and derived tumors.
Two monoclonal antibodies (LN-1, LN-2) reactive with B lymphocytes in B5 formalin-fixed, paraffin-embedded tissue sections have been produced by utilizing cell extracts from pokeweed mitogen-stimulated peripheral blood lymphocytes and diffuse histiocytic lymphoma SU-DHL-4 cells, respectively. Both monoclonal antibodies were initially identified by indirect immunofluorescence screening techniques on paraformaldehyde-acetone-fixed cell preparations. Specificity screens with 36 well-characterized human lymphoma and leukemia cell lines showed that both LN-1 and LN-2 stained cell lines of B cell lineage but were unreactive with those of T cell or, with one exception, myeloid derivation. Null cell acute lymphoblastic leukemia cell lines were found to be LN-2+ but LN-1-. The B cell specificity of these reagents was confirmed on 15 lymphoma and 17 leukemia biopsy specimens by using indirect immunofluorescence techniques. Immunoperoxidase staining of sections from B5-fixed, paraffin-embedded human lymphoid tissues showed that LN-1 bound to the cell membrane and cytoplasm of germinal center cells whereas LN-2 stained the nuclear membrane and cytoplasm of germinal center and mantle zone B lymphocytes as well as interfollicular histiocytes and thymic medullary dendritic cells. Both monoclonal antibodies failed to stain cortical thymocytes, lymph node T cells, and peripheral blood T and myeloid cells. Immunoperoxidase staining of 20 nonlymphoid human organs and tissues revealed that LN-1 reacted positively with red blood cell precursors of the bone marrow, ciliated epithelial cells of the bronchus, distal tubular cells of the kidney, and ductal cells from several organs including the breast and prostate. In contrast, LN-2 was unreactive with all human nonlymphoid organs and tissues including the bone marrow. Indirect immunofluorescence staining of a panel of 26 solid tumor cells lines showed that LN-1 was reactive with the majority of epithelium-derived cell lines, glioblastomas, and astrocytomas but was unreactive with neuroblastomas, small cell carcinoma of the lung, and sarcomas. LN-2 was unreactive with 25 of 26 of the solid tumor cell lines by these techniques. Immunobiochemical studies have shown that LN-1 recognizes a cell surface sialoantigen whereas LN-2 is directed against a 35,000 dalton nuclear membrane protein. Because of their high specificity for B cell tumors and their ability to stain B5-fixed, paraffin-embedded tissues, LN-1 and LN-2 are useful reagents for the diagnosis and classification of the human lymphomas and leukemias.


Gene expression and functional annotation of the human and mouse choroid plexus epithelium.
The choroid plexus epithelium (CPE) is a lobed neuro-epithelial structure that forms the outer blood-brain barrier. The CPE protrudes into the brain ventricles and produces the cerebrospinal fluid (CSF), which is crucial for brain homeostasis. Malfunction of the CPE is possibly implicated in disorders like Alzheimer disease, hydrocephalus or glaucoma. To study human genetic diseases and potential new therapies, mouse models are widely used. This requires a detailed knowledge of similarities and differences in gene expression and functional annotation between the species. The aim of this study is to analyze and compare gene expression and functional annotation of healthy human and mouse CPE. We performed 44k Agilent microarray hybridizations with RNA derived from laser dissected healthy human and mouse CPE cells. We functionally annotated and compared the gene expression data of human and mouse CPE using the knowledge database Ingenuity. We searched for common and species specific gene expression patterns and function between human and mouse CPE. We also made a comparison with previously published CPE human and mouse gene expression data. Overall, the human and mouse CPE transcriptomes are very similar. Their major functionalities included epithelial junctions, transport, energy production, neuro-endocrine signaling, as well as immunological, neurological and hematological functions and disorders. The mouse CPE presented two additional functions not found in the human CPE: carbohydrate metabolism and a more extensive list of (neural) developmental functions. We found three genes specifically expressed in the mouse CPE compared to human CPE, being ACE, PON1 and TRIM3 and no human specifically expressed CPE genes compared to mouse CPE. Human and mouse CPE transcriptomes are very similar, and display many common functionalities. Nonetheless, we also identified a few genes and pathways which suggest that the CPE between mouse and man differ with respect to transport and metabolic functions.


Quantitative MALDI tandem mass spectrometric imaging of cocaine from brain tissue with a deuterated internal standard.
Mass spectrometric imaging (MSI) is an analytical technique used to determine the distribution of individual analytes within a given sample. A wide array of analytes and samples can be investigated by MSI, including drug distribution in rats, lipid analysis from brain tissue, protein differentiation in tumors, and plant metabolite distributions. Matrix-assisted laser desorption/ionization (MALDI) is a soft ionization technique capable of desorbing and ionizing a large range of compounds, and it is the most common ionization source used in MSI. MALDI mass spectrometry (MS) is generally considered to be a qualitative analytical technique because of significant ion-signal variability. Consequently, MSI is also thought to be a qualitative technique because of the quantitative limitations of MALDI coupled with the homogeneity of tissue sections inherent in an MSI experiment. Thus, conclusions based on MS images are often limited by the inability to correlate ion signal increases with actual concentration increases. Here, we report a quantitative MSI method for the analysis of cocaine (COC) from brain tissue using a deuterated internal standard (COC-d(3)) combined with wide-isolation MS/MS for analysis of the tissue extracts with scan-by-scan COC-to-COC-d(3) normalization. This resulted in significant improvements in signal reproducibility and calibration curve linearity. Quantitative results from the MSI experiments were compared with quantitative results from liquid chromatography (LC)-MS/MS results from brain tissue extracts. Two different quantitative MSI techniques (standard addition and external calibration) produced quantitative results comparable to LC-MS/MS data. Tissue extracts were also analyzed by MALDI wide-isolation MS/MS, and quantitative results were nearly identical to those from LC-MS/MS. These results clearly demonstrate the necessity for an internal standard for quantitative MSI experiments.


Photovoltaic and electrocatalytic properties of novel ball-type phthalocyanines bridged with four dicumarol.
The new ball-type metallo bisphthalocyanines (Co(2)Pc(2) and Zn(2)Pc(2)) were synthesized from the corresponding [4,4'-bis(dicoumaroylphthalonitrile)] which can be obtained from the reaction of 3,3'-methylenebis(4-hydroxy-2H-chromen-2-one) and 4-nitrophthalonitrile. The structures of the newly synthesized compounds have been confirmed and characterized by elemental analysis, UV/Vis, IR and (1)H NMR spectroscopies and MALDI-TOF mass spectrometry. Solar cells of the configuration ITO/Co(2)Pc(2)/C60/Al and ITO/Zn(2)Pc(2)/C60/Al were fabricated. The effect of the thickness of the active Pc layer--the thickness of the Pc layer was varied from 15 to 80 nm--on solar cells parameters has been investigated. A nearly thickness independent open circuit voltage was observed in both structures. The maximum photovoltaic conversion efficiency, short circuit current and fill factor were observed in ITO/Zn(2)Pc(2)/C60/Al cell with 80 nm Pc layer to be 0.255%, 1 mA cm(-2) and 0.38, respectively. The redox properties of the ball-type complexes were investigated by cyclic voltammetry, controlled-potential coulometry and in situ spectroelectrochemistry in DMSO-TBAP. The electrochemical measurements showed that the complexes form ring-based and/or metal-based mixed-valence species, due to the remarkable intramolecular interactions between the two metal phthalocyanine units. The Vulcan XC-72(VC)/Nafion(Nf)/Co(2)Pc(2) modified glassy carbon electrode showed much higher catalytic performance towards oxygen reduction, compared to the VC/Nf/Zn(2)Pc(2) modified one. It was found that the VC/Nf/Co(2)Pc(2) catalyst is nearly insensitive to the presence of methanol. In the presence of 1 M methanol in the electrolyte, the catalytic performance of the Co(2)Pc(2)-based catalyst in oxygen reduction was much better than that of the Pt-based one. Thus, it was shown that the VC/Nf/Co(2)Pc(2) catalyst can be a good alternative to VC/Nf/Pt as a cathode catalyst in direct methanol fuel cells.


Mechanics on myocardium deficient in the N2B region of titin: the cardiac-unique spring element improves efficiency of the cardiac cycle.
Titin (also known as connectin) is an intrasarcomeric muscle protein that functions as a molecular spring and generates passive tension upon muscle stretch. The N2B element is a cardiac-specific spring element within titin's extensible region. Our goal was to study the contribution of the N2B element to the mechanical properties of titin, particularly its hypothesized role in limiting energy loss during repeated stretch (diastole)-shortening (systole) cycles of the heart. We studied energy loss by measuring hysteresis from the area between the stretch and release passive force-sarcomere length curves and used both wild-type (WT) mice and N2B knockout (KO) mice in which the N2B element has been deleted. A range of protocols was used, including those that mimic physiological loading conditions. KO mice showed significant increases in hysteresis. Most prominently, in tissue that had been preconditioned with a physiological stretch-release protocol, hysteresis increased significantly from 320 ± 46 pJ/mm(2)/sarcomere in WT to 650 ± 94 pJ/mm(2)/sarcomere in N2B KO myocardium. These results are supported by experiments in which oxidative stress was used to mechanically inactivate portions of the N2B-Us of WT titin through cysteine cross-linking. Studies on muscle from which the thin filaments had been extracted (using the actin severing protein gelsolin) showed that the difference in hysteresis between WT and KO tissue cannot be explained by filament sliding-based viscosity. Instead the results suggest that hysteresis arises from within titin and most likely involves unfolding of immunoglobulin-like domains. These studies support that the mechanical function of the N2B element of titin includes reducing hysteresis and increasing the efficiency of the heart.


Programming microbes using pulse width modulation of optical signals.
Cells transmit and receive information via signalling pathways. A number of studies have revealed that information is encoded in the temporal dynamics of these pathways and has highlighted how pathway architecture can influence the propagation of signals in time and space. The functional properties of pathway architecture can also be exploited by synthetic biologists to enable precise control of cellular physiology. Here, we characterised the response of a bacterial light-responsive, two-component system to oscillating signals of varying frequencies. We found that the system acted as a low-pass filter, able to respond to low-frequency oscillations and unable to respond to high-frequency oscillations. We then demonstrate that the low-pass filtering behavior can be exploited to enable precise control of gene expression using a strategy termed pulse width modulation (PWM). PWM is a common strategy used in electronics for information encoding that converts a series of digital input signals to an analog response. We further show how the PWM strategy extends the utility of bacterial optogenetic control, allowing the fine-tuning of expression levels, programming of temporal dynamics, and control of microbial physiology via manipulation of a metabolic enzyme.


Influence of insulin antibodies on pharmacokinetics and bioavailability of recombinant human and highly purified beef insulins in insulin dependent diabetics.
Sixteen insulin dependent diabetics of long standing, with undetectable fasting plasma C peptide concentrations, and eight non-diabetic controls were each infused intravenously with biosynthetic human and highly purified beef insulin (1 mU/kg/min) while euglycaemia was maintained by a Biostator. No difference was observed between the two insulins in respect of insulin pharmacokinetics or biological action. The diabetics showed appreciable insulin resistance, manifested by a 40% reduction in the rate of insulin mediated glucose disposal, which was unrelated to the presence of insulin antibodies. Insulin binding antibodies, however, increased insulin's clearance rate and distribution space and prolonged its pharmacological and biological half lives. The rate at which insulin action was lost, after an intravenous infusion, was more rapid in diabetics without insulin antibody binding than in controls. In respect of their influence on insulin pharmacokinetics, moderate concentrations of insulin antibodies may be of positive advantage to all diabetics without endogenous insulin secretion and are not responsible for the insulin resistance of type 1 diabetes.


Omega-3 eicosapentaenoic acid in bipolar depression: report of a small open-label study.
Epidemiologic studies have suggested that consumption of cold water fish oils may have some protective function against depression. This proposition is supported by a series of biochemical and pharmacologic studies that have suggested that fatty acids may modulate neurotransmitter metabolism and cell signal trans-duction in humans and that abnormalities in fatty acid and eicosanoid metabolism may play a causal role in depression. Aware of the critical need for antidepression treatments that might not carry the risk of precipitating a manic episode in bipolar patients, we decided to conduct an open-label add-on trial of eicosapentaenoic acid (EPA) in bipolar depression. Twelve bipolar I outpatients with depressive symptoms diagnosed by DSM-IV were treated with 1.5 to 2 g/day of the omega-3 fatty acid EPA for up to 6 months. The study was conducted between September 2001 and January 2003. Eight of the 10 patients who completed at least 1 month of follow-up achieved a 50% or greater reduction in Hamilton Rating Scale for Depression scores within 1 month. No patients developed hypomania or manic symptoms. No significant side effects were reported. This study is limited both by the open-label design and by the small sample size. As in all previous reported studies, patients in this study were treated in an outpatient setting, so that the most severely depressed bipolar patients (requiring hospitalization) are not represented. Although the ultimate utility of omega-3 fatty acids in bipolar depression is still an open question, we believe that these initial results are encouraging, especially for mild to moderate bipolar depression, and justify the continuing exploration of its use.


Not supposed to feel this: traditional masculinity in psychotherapy with male veterans returning from Afghanistan and Iraq.
Traditional masculine socialization presents challenges in psychotherapy, for example, by decreasing the likelihood of help-seeking and by making emotion-laden content more difficult to address. While this has been established in civilian populations, more intense forms of masculine socialization found in military settings may amplify such issues in male veteran populations. Male veterans returning from and Afghanistan (OEF) and Iraq (OIF) exhibit strong traditional masculine socialization and generally present in a unique manner. It is posited that OEF/OIF male veterans' unique presentation is in large part because of an interaction between high degrees of endorsement of traditional masculine gender role norms, relative youth, recency of distressing events, and recent experience in the social context of the military where traditional masculinity is reinforced. The impact of these variables on the psychotherapeutic process for male OEF/OIF veterans is significant and likely adds to ambivalence about change and increases dropout from psychotherapy. Modifications of traditional psychotherapeutic approaches designed to address traditional masculine gender role norms and their many interactions with other variables are discussed.


Plasmon Rulers as a Probe for Real-Time Microsecond Conformational Dynamics of Single Molecules.
Biopolymers such as DNA, RNA, and proteins exploit conformational changes to modulate their function. Although state-of-the-art single-molecule approaches enable identification of conformational states, the transition path and metastable intermediates often remain elusive because they occur on microsecond time scales. Here we introduce a method to probe conformational dynamics with microsecond integration times based on a heterodimer of plasmonic particles. By combining Brownian dynamics and electromagnetic simulations, we find that integration times of 1 μs can be routinely achieved, providing the capability to identify short-lived intermediates and transition paths at the single-molecule level in real-time. Importantly, plasmon rulers require no specialized equipment but can be probed on existing fluorescence microscopes equipped with a fast camera. The approach combines the advantages of fluorescent probes (zero-force, parallelization) and mechanical probes such as optical tweezers (continuous microsecond integration times). They offer a unique opportunity to study conformational dynamics and compare measurements to full-atom simulations, where computational demands limit the simulation time.


A comparison of dry powder inhaler dose delivery characteristics using a power criterion.
A novel method is described that compares the in vitro drug delivery characteristics of different dry powder inhalers (DPIs) based on the power supplied by the air flow. Power is defined as the rate at which work is done by the air flow and is equal to the product of pressure gradient and air flow rate through the inhaler. Tests were conducted to compare the emitted dose and fine particle fraction of the total dose (FPFTD) of the Diskus, Diskhaler, and a proprietary powder dispersion device (PDD) containing fluticasone propionate/lactose powder blends. Three power levels were selected: 1, 2, and 3 W, representing a range of inspiratory efforts. The Diskus and PDD were independent of power, delivering consistent doses and fine particle fractions over the range of powers evaluated. Diskhaler was dependent on power, the emitted dose and FPFTD significantly decreasing at the lower power level (1W) (p < 0.005). Diskus and Diskhaler are trademarks of the Glaxo Wellcome group of companies.


Polymeric nanovesicles as simultaneous delivery platforms with doxorubicin conjugation and elacridar encapsulation for enhanced treatment of multidrug-resistant breast cancer.
Multidrug resistance (MDR) is one of the major obstacles hindering the successful chemotherapy of cancer. Overexpression of drug efflux transporters such as P-glycoprotein (P-gp) is an important factor responsible for MDR. In this study, a novel copolymer methoxy-poly(ethylene glycol)-poly[(N-(6-hydroxyhexyl)-g-doxorubicin-l-aspartamide)-(β-benzyl-l-aspartate)] (mPEG-P[Asp(HPA-g-DOX)-BLA)] was synthesized and utilized to assemble into nanovesicles with hydrophilic P-gp inhibitor elacridar hydrochloride (Ela) encapsulated into the aqueous lumen. Doxorubicin (DOX) was covalently conjugated onto the polymer chain via a pH-cleavable amide linkage, leading to a pH responsive DOX release as well as disintegration of the nanovesicles in the lysosome of tumor cells. In vitro studies demonstrated that the DOX and Ela co-delivered nanovesicles showed superior cytotoxicity and enhanced anti-tumor properties as compared to single DOX-delivery nanosystems in MCF-7/ADR cancer, which was attributed to the P-gp bioactivity inhibition as investigated by a cell immunofluorescence assay. In vivo studies showed that the polymeric nanovesicles effectively accumulated at the tumor site and the co-delivered DOX and Ela effectively suppressed the MCF-7/ADR tumor growth. All the results indicated that the acid-liable nanovesicles had a synergistic effect to enhance antitumor efficacy for multidrug-resistant breast cancer treatment.


Dynamics of parabolic pulses in an ultrafast fiber laser.
We report experimental and numerical results on the dynamics and propagation of parabolic pulses in a passively mode-locked ytterbium-doped fiber laser. Experimental data and numerical simulations are shown to match. Particular importance is attached to the pulse-shaping process in the different sections of the resonator. The paramount role of the gain fiber and saturable absorber in the laser is explicated.


A possible link between food and mood: dietary impact on gut microbiota and behavior in BALB/c mice.
Major depressive disorder is a debilitating disease in the Western World. A western diet high in saturated fat and refined sugar seems to play an important part in disease development. Therefore, this study is aimed at investigating whether saturated fat or sucrose predisposes mice to develop behavioral symptoms which can be interpreted as depression-like, and the possible influence of the gut microbiota (GM) in this. Fourty-two mice were randomly assigned to one of three experimental diets, a high-fat, a high-sucrose or a control diet for thirteen weeks. Mice on high-fat diet gained more weight (p = 0.00009), displayed significantly less burrowing behavior than the control mice (p = 0.034), and showed decreased memory in the Morris water maze test compared to mice on high-sucrose diet (p = 0.031). Mice on high-sucrose diet burrowed less goal-oriented, showed greater latency to first bout of immobility in the forced swim test when compared to control mice (p = 0.039) and high-fat fed mice (p = 0.013), and displayed less anxiety than mice on high-fat diet in the triple test (p = 0.009). Behavioral changes were accompanied by a significant change in GM composition of mice fed a high-fat diet, while no difference between diet groups was observed for sucrose preferences, LPS, cholesterol, HbA1c, BDNF and the cytokines IL-1α, IL-1β, IL-6, IL-10, IL-12(p70), IL-17 and TNF-α. A series of correlations was found between GM, behavior, BDNF and inflammatory mediators. In conclusion, the study shows that dietary fat and sucrose affect behavior, sometimes in opposite directions, and suggests a possible association between GM and behavior.


Reproducibility of performance measures of physical function based on the BASFI, in ankylosing spondylitis.
To establish the test-retest reproducibility of performance measures of physical function based on the Bath Ankylosing Spondylitis Functional Index (BASFI) questionnaire in patients with AS. Data were obtained from 65 AS patients. They were tested on two occasions by one assessor with a 1-week interval. Physical function was assessed via eight performance measures based on items used in the BASFI questionnaire, representing activities of daily life, which AS patients frequently report to be problematic. For each activity, a performance score was determined. Pain and exertion were measured using a 10-cm horizontal visual analogue scale (VAS) and Borg's modified scale, respectively. Test-retest reproducibility was assessed for all measurements using intraclass correlation coefficients (ICCs) and by calculating the standard error of measurement (SEM). Adequate intrarater reliability was found. For performance scores, ICCs ranged from 0.73 to 0.96. Measurements of exertion and pain also showed adequate intrarater reliability, with the exception of one performance measure, namely the test for the ability to look over one's shoulder. For this test, the ICCs were 0.66 and 0.69 for exertion and pain, respectively. The remaining ICCs for exertion ranged from 0.71 to 0.88 and for pain from 0.74 to 0.83. The SEM for performance scores ranged from 4 to 9% of the observed score. The SEM for exertion ranged from 8 to 11% and for pain from 10 to 15%. Performance measures of physical function based on the BASFI questionnaire have adequate to excellent test-retest reproducibility. Due to the presence of measurement error, measurements are accurate for group assessment; repeated measurements are advised for an adequate assessment of individual patients.


Fine mapping and characterization of the Rous sarcoma virus Pr76gag late assembly domain.
The p2 region of the Rous sarcoma virus (RSV) Gag polyprotein contains an assembly domain, which is required late in replication for efficient budding of virus-like particles from cells (J. W. Wills, C. E. Cameron, C. B. Wilson, Y. Xiang, R. P. Bennett, and J. Leis, J. Virol. 68:6605-6618, 1994). This domain, referred to as the L domain, was previously mapped to the 11 amino acids of p2b. Through the analysis of a series of deletion and substitution mutations, the L domain has now been fine mapped to a highly conserved amino acid sequence, PPPPYV of p2b. Sequences flanking PPPPYV motif can be deleted without any effect on budding. Defects caused by L-domain deletions can be rescued by placing a wild-type copy of the sequence at several other positions in RSV Gag. A proline-rich P(S/T)APP motif is found in many retroviral Gag polyproteins; the motif found in the p6 region of human immunodeficiency virus type 1 has been implicated in late functions of the virus. Substitution of the RSV L domain with this motif in a 10-amino-acid sequence derived from visna leukemia virus results in wild-type release of virus particles from cells. In contrast, the slightly different sequences from Gibbon ape leukemia virus, Moloney leukemia virus, PSAPP alone, or a proline-rich SH3 binding sequence do not efficiently rescue RSV L-domain mutations.


ABCB5 is a limbal stem cell gene required for corneal development and repair.
Corneal epithelial homeostasis and regeneration are sustained by limbal stem cells (LSCs), and LSC deficiency is a major cause of blindness worldwide. Transplantation is often the only therapeutic option available to patients with LSC deficiency. However, while transplant success depends foremost on LSC frequency within grafts, a gene allowing for prospective LSC enrichment has not been identified so far. Here we show that ATP-binding cassette, sub-family B, member 5 (ABCB5) marks LSCs and is required for LSC maintenance, corneal development and repair. Furthermore, we demonstrate that prospectively isolated human or murine ABCB5-positive LSCs possess the exclusive capacity to fully restore the cornea upon grafting to LSC-deficient mice in xenogeneic or syngeneic transplantation models. ABCB5 is preferentially expressed on label-retaining LSCs in mice and p63α-positive LSCs in humans. Consistent with these findings, ABCB5-positive LSC frequency is reduced in LSC-deficient patients. Abcb5 loss of function in Abcb5 knockout mice causes depletion of quiescent LSCs due to enhanced proliferation and apoptosis, and results in defective corneal differentiation and wound healing. Our results from gene knockout studies, LSC tracing and transplantation models, as well as phenotypic and functional analyses of human biopsy specimens, provide converging lines of evidence that ABCB5 identifies mammalian LSCs. Identification and prospective isolation of molecularly defined LSCs with essential functions in corneal development and repair has important implications for the treatment of corneal disease, particularly corneal blindness due to LSC deficiency.


Rifamide in acute cholecystitis and biliary surgery.
Determination of the minimum inhibitory concentrations of rifamide necessary to inhibit organisms isolated from the biliary tract showed that the organisms were almost invariably sensitive to concentrations which are readily attainable in the biliary tract. Three cases of severe acute inflammation of the biliary tract were treated and this led to rapid clinical improvement. In 61 patients undergoing biliary surgery a random group was given rifamide 150 mg twice daily, beginning 24 hours before operation and continuing for three days afterwards. In the untreated group eight patients had infected bile at operation and five subsequently developed a wound infection. In the rifamide group three had infected bile at operation and only one developed a wound infection. A similar number of postoperative chest infections occurred in each group of patients. There is some evidence of reduction in length of hospital stay in the treated patients.


Estimating the duration of Plasmodium falciparum infection from trials of indoor residual spraying.
We reviewed the use of simple mathematical models to estimate the duration of Plasmodium falciparum infection after transmission has been interrupted. We then fit an exponential decay model to repeated cross-sectional survey data collected from three historical trials of indoor residual spraying against malaria: one from two contiguous districts in Tanzania-Kenya (Pare Taveta) carried out in 1954, the others in West Papua (1953), and the Garki project in northern Nigeria (1972-1973). A cross-sectional analysis of these datasets gave overall estimates of 602 days (95% confidence interval [CI] = 581-625) for the infection duration in Pare Taveta, 734 days (95% CI = 645-849) in West Papua, and 1,329 days (95% CI =1,193-1,499) for Garki. These estimates are much greater than the most widely quoted figures for the duration of untreated P. falciparum infections. Although these may be exaggerated because some reinfections occurred despite intensive vector control, prevalence was still decreasing when all these projects ended. Longitudinal survival analysis of the Garki data gave much shorter estimates of duration (186 days, 95% CI = 181-191), but effects of imperfect detection of parasites by microscopy severely bias these estimates. Estimates of infection duration for different age groups showed considerable variation but no general age trend. There was also no clear relationship between malaria endemicity and infection duration. Analyses of successive sampling from the same individuals with parasite typing are needed to obtain more reliable estimates of infection duration in endemic areas. Periods of several years may be required to evaluate long-term effects of interventions on malaria prevalence.


Thoracic duct drainage before and after cadaveric kidney transplantation.
Twenty-seven consecutive recipients of cadaveric kidneys, including five with pre-existing warm cytotoxic antibodies, were treated with thoracic duct drainage before and after transplantation. Fourteen patients who had lymph drainage for 26 to 58 days before transplantation had minor cytotoxic antibody responses after grafting, even if the antibodies had been present before therapy. Only one of the 14 recipients had any rejection during the follow-up periods of one to six months. There were two deaths. The 13 patients pretreated for 17 to 23 days exhibited stronger cytotoxic antibody responsiveness, and five of these recipients had significant rejections of which four were reversible. One of the latter 13 patients died. These clinical and immunologic studies have established the value and have defined the appropriate timing of preoperative thoracic duct drainage in kidney transplantation. They have also directed attention to the rationale andthe probable value of using other immunosuppressive methods for preparatory host conditioning instead of beginning such therapy at the time of transplantation.


Emotion modulation in PTSD: Clinical and neurobiological evidence for a dissociative subtype.
In this article, the authors present evidence regarding a dissociative subtype of PTSD, with clinical and neurobiological features that can be distinguished from nondissociative PTSD. The dissociative subtype is characterized by overmodulation of affect, while the more common undermodulated type involves the predominance of reexperiencing and hyperarousal symptoms. This article focuses on the neural manifestations of the dissociative subtype in PTSD and compares it to those underlying the reexperiencing/hyperaroused subtype. A model that includes these two types of emotion dysregulation in PTSD is described. In this model, reexperiencing/hyperarousal reactivity is viewed as a form of emotion dysregulation that involves emotional undermodulation, mediated by failure of prefrontal inhibition of limbic regions. In contrast, the dissociative subtype of PTSD is described as a form of emotion dysregulation that involves emotional overmodulation mediated by midline prefrontal inhibition of the same limbic regions. Both types of modulation are involved in a dynamic interplay and lead to alternating symptom profiles in PTSD. These findings have important implications for treatment of PTSD, including the need to assess patients with PTSD for dissociative symptoms and to incorporate the treatment of dissociative symptoms into stage-oriented trauma treatment.


Volume guarantee versus high-frequency ventilation: lung inflammation in preterm infants.
Appropriate ventilation together with improvement of clinical care of premature babies can contribute to reducing lung inflammation, known to represent the "primum movens" of bronchopulmonary dysplasia (BPD). High-frequency oscillatory ventilation (HFOV) and volume-guarantee (VG) ventilation are effective in the treatment of neonatal respiratory distress syndrome (RDS). To assess the potential of HFOV and VG to prevent BPD in the acute phase of RDS, by a randomised clinical study evaluating lung inflammation in premature infants. Forty infants (gestational age 25-32 weeks) with RDS were assigned to assist-control ventilation plus VG (Vt = 5 ml/kg) or HFOV (both with a Dräger Babylog 8000 plus ventilator). Levels of interleukin (IL) 6, IL8 and tumour necrosis factor were determined in tracheal aspirate on days 1, 3 and 7 of life. In the HFOV group IL6 levels were significantly higher on day 3 (0.5 (0.2) vs assisted-control ventilation plus VG group 0.1 (0.2) ng/ml) and oxygen dependency was significantly longer (36 (23) vs assisted-control ventilation plus VG group 19 (11) days). VG ventilation is an effective lung-protective strategy to be used in acute RDS, inducing a lower expression of early inflammation markers than HFOV. Whether the use of this initial ventilatory strategy contributes to the prevention of BPD requires further studies.


Response of SMRT (silencing mediator of retinoic acid and thyroid hormone receptor) and N-CoR (nuclear receptor corepressor) corepressors to mitogen-activated protein kinase kinase kinase cascades is determined by alternative mRNA splicing.
The SMRT (silencing mediator of retinoic acid and thyroid hormone receptor) and N-CoR (nuclear receptor corepressor) corepressors are important mediators of transcriptional repression by nuclear hormone receptors. SMRT is regulated by MAPK kinase kinase (MAPKKK) cascades that induce its release from its receptor partners, its export from nucleus to cytoplasm, and derepression of target gene expression. Intriguingly, the otherwise closely related N-CoR is refractory to MAPKKK signaling under the same conditions. However, both SMRT and N-CoR are expressed as a series of alternatively spliced protein variants differing in structure and function. We have now characterized the impact of this alternative mRNA splicing on the corepressor response to MAPKKK signaling. Whereas the SMRTalpha, SMRTtau, and SMRTsp2 splice variants are released from their nuclear receptor partners in response to MAPKKK activation, the SMRTsp18 variant, which resembles N-CoR in its overall molecular architecture, is relatively refractory to this kinase-induced release. Alternative splicing of N-CoR, in contrast, had only minimal effects on the resistance of this corepressor to MAPKKK inhibition. Notably, all of the SMRT splice variants examined redistributed from nucleus to cytoplasm in response to MAPKKK cascade signaling, but none of the N-CoR splice variants did so. Different tiers of the MAPKKK cascade hierarchy contributed to these different aspects of corepressor regulation, with MAP/ERK kinase kinase 1 and MAP/ERK kinase 1 regulating subcellular redistribution and ERK2 regulating nuclear receptor-corepressor interaction. We conclude that cells can customize their transcriptional response to MAPKKK cascade signaling by selective expression of the SMRT or N-CoR locus, by selective utilization of a specific corepressor splice variant, and by selective exploitation of specific tiers of the MAPK cascade.


Developmental and neurochemical features of cholinergic neurons in the murine cerebral cortex.
The existence and role of intrinsic cholinergic cells in the cerebral cortex is controversial, because of their variable localization and morphology in different mammalian species. We have applied choline acetyltransferase (ChAT) immunocytochemistry to study the distribution of cholinergic neurons in the murine cerebral cortex, in the adult and during postnatal development. For more precise neurochemical identification of these neurons, the possible colocalization of ChAT with different markers of cortical neuronal populations has been analyzed by confocal microscopy. This method was also used to verify the relationship between cholinergic cells and cortical microvessels. ChAT positive cells appeared at the end of the first postnatal week. Their density dramatically increased at the beginning of the second postnatal week, during which it remained higher than in perinatal and adult stages. In the adult neocortex, cholinergic neurons were particularly expressed in the somatosensory area, although their density was also significant in visual and auditory areas. ChAT positive cells tended to be scarce in other regions. They were mainly localized in the supragranular layers and displayed a fusiform/bipolar morphology. The colocalization of ChAT with pyramidal neuron markers was negligible. On the other hand, more than half of the cholinergic neurons contained calretinin, but none of them expressed parvalbumin or calbindin. However, only a fraction of the ChAT positive cells during development and very few in adulthood turned out to be GABAergic, as judged from expression of GABA and its biosynthetic enzymes GAD67/65. Consistently, ChAT showed no localization with interneurons expressing green fluorescent protein under control of the GAD67 promoter in the adult neocortex. Finally, the cortical cholinergic cells often showed close association with the microvessel walls, as identified with the gliovascular marker aquaporin 4, supporting previous hypotheses on the role of cholinergic cells in modulating the cortical microcirculation. Our results show that the development of the intracortical cholinergic system accompanies the cortical rearrangements during the second postnatal week, a crucial stage for the establishment of cortical cytoarchitecture and for synaptogenesis. Although intrinsic ChAT positive cells usually expressed calretinin, they displayed a variable GABAergic phenotype depending on marker and on cortical developmental stage.


Overestimates of survival after HAART: implications for global scale-up efforts.
Monitoring the effectiveness of global antiretroviral therapy scale-up efforts in resource-limited settings is a global health priority, but is complicated by high rates of losses to follow-up after treatment initiation. Determining definitive outcomes of these lost patients, and the effects of losses to follow-up on estimates of survival and risk factors for death after HAART, are key to monitoring the effectiveness of global HAART scale-up efforts. A cohort study comparing clinical outcomes and risk factors for death after HAART initiation as reported before and after tracing of patients lost to follow-up was conducted in Botswana's National Antiretroviral Therapy Program. 410 HIV-infected adults consecutively presenting for HAART were evaluated. The main outcome measures were death or loss to follow-up within the first year after HAART initiation. Of 68 patients initially categorized as lost, over half (58.8%) were confirmed dead after tracing. Patient tracing resulted in reporting of significantly lower survival rates when death was used as the outcome and losses to follow-up were censored [1-year Kaplan Meier survival estimate 0.92 (95% confidence interval, 0.88-0.94 before tracing and 0.83 (95% confidence interval, 0.79-0.86) after tracing, log rank P<0.001]. In addition, a significantly increased risk of death after HAART among men [adjusted hazard ratio 1.74 (95% confidence interval, 1.05-2.87)] would have been missed had patients not been traced [adjusted hazard ratio 1.41 (95% confidence interval, 0.65-3.05)]. Due to high rates of death among patients lost to follow-up after HAART, survival rates may be inaccurate and important risk factors for death may be missed if patients are not actively traced. Patient tracing and uniform reporting of outcomes after HAART are needed to enable accurate monitoring of global HAART scale-up efforts.


Effects of chronic morphine administration on the reproductive system of the male rat.
The effects of chronic morphine administration on a variety of rat organs were examined. Only the prostates and seminal vesicles were affected by a 3-day period of morphine pellet implantation. In these organs, particularly the seminal vesicles, morphine produced a marked decrease in both the wet and dry tissue weight and a reduction in their secretory activity. This apparent morphine-induced atrophy of these important accessory sex organs was not due to a change in the nutritional status of the animals but rather appeared to be associated with a pronounced (87 percent) reduction in the serum testosterone levels of morphine-treated rats when compared to control levels. The effect of morphine on the secondary sex organs was reversed by withdrawing the animals from morphine administration for a period of 7 days. These data suggest that the role of physiological and endocrinological factors in the narcotic addict's sexual difficulties be examined.


Development and validation of a knowledge test for health professionals regarding lifestyle modification.
We wanted to develop and validate a test that assesses the knowledge and practices of health professionals (HPs) with regard to the role of nutrition, physical activity, and smoking cessation (lifestyle modification) in chronic diseases of lifestyle. A descriptive cross-sectional validation study was carried out. The validation design consisted of two phases, namely 1) test planning and development and 2) test evaluation. The study sample consisted of five groups of HPs: dietitians, dietetic interns, general practitioners, medical students, and nurses. The overall response rate was 58%, resulting in a sample size of 186 participants. A test was designed to evaluate the knowledge and practices of HPs. The test was first evaluated by an expert group to ensure content, construct, and face validity. Thereafter, the questionnaire was tested on five groups of HPs to test for criterion validity. Internal consistency was evaluated by Cronbach's alpha. An expert panel ensured content, construct, and face validity of the test. Groups with the most training and exposure to nutrition (dietitians and dietetic interns) had the highest group mean score, ranging from 61% to 88%, whereas those with limited nutrition training (general practitioners, medical students, and nurses) had significantly lower scores, ranging from 26% to 80%. This result demonstrated criterion validity. Internal consistency of the overall test demonstrated a Cronbach's alpha of 0.99. Most HPs identified the mass media as their main source of information on lifestyle modification. These HPs also identified lack of time, lack of patient compliance, and lack of knowledge as barriers that prevent them from providing counseling on lifestyle modification. The results of this study showed that this test instrument identifies groups of health professionals with adequate training (knowledge) in lifestyle modification and those who require further training (knowledge).


Socioeconomic hierarchy and health gradient in Europe: the role of income inequality and of social origins.
Health inequalities reflect multidimensional inequality (income, education, and other indicators of socioeconomic position) and vary across countries and welfare regimes. To which extent there is intergenerational transmission of health via parental socioeconomic status has rarely been investigated in comparative perspective. The study sought to explore if different measures of stratification produce the same health gradient and to which extent health gradients of income and of social origins vary with level of living and income inequality. A total of 299,770 observations were available from 18 countries assessed in EU-SILC 2005 and 2011 data, which contain information on social origins. Income inequality (Gini) and level of living were calculated from EU-SILC. Logit rank transformation provided normalized inequalities and distributions of income and social origins up to the extremes of the distribution and was used to investigate net comparable health gradients in detail. Multilevel random-slope models were run to post-estimate best linear unbiased predictors (BLUPs) and related standard deviations of residual intercepts (median health) and slopes (income-health gradients) per country and survey year. Health gradients varied across different measures of stratification, with origins and income producing significant slopes after controls. Income inequality was associated with worse average health, but income inequality and steepness of the health gradient were only marginally associated. Linear health gradients suggest gains in health per rank of income and of origins even at the very extremes of the distribution. Intergenerational transmission of status gains in importance in countries with higher income inequality. Countries differ in the association of income inequality and income-related health gradient, and low income inequality may mask health problems of vulnerable individuals with low status. Not only income inequality, but other country characteristics such as familial orientation play a considerable role in explaining steepness of the health gradient.


Prediction of manifest Huntington's disease with clinical and imaging measures: a prospective observational study.
Although the association between cytosine-adenine-guanine (CAG) repeat length and age at onset of Huntington's disease is well known, improved prediction of onset would be advantageous for clinical trial design and prognostic counselling. We compared various measures for tracking progression and predicting conversion to manifest Huntington's disease. In this prospective observational study, we assessed the ability of 40 measures in five domains (motor, cognitive, psychiatric, functional, and imaging) to predict time to motor diagnosis of Huntington's disease, accounting for CAG repeat length, age, and the interaction of CAG repeat length and age. Eligible participants were individuals from the PREDICT-HD study (from 33 centres in six countries [USA, Canada, Germany, Australia, Spain, UK]) with the gene mutation for Huntington's disease but without a motor diagnosis (a rating below 4 on the diagnostic confidence level from the 15-item motor assessment of the Unified Huntington's Disease Rating Scale). Participants were followed up between September, 2002, and July, 2014. We used joint modelling of longitudinal and survival data to examine the extent to which baseline and change of measures analysed separately was predictive of CAG-adjusted age at motor diagnosis. 1078 individuals with a CAG expansion were included in this analysis. Participants were followed up for a mean of 5·1 years (SD 3·3, range 0·0-12·0). 225 (21%) of these participants received a motor diagnosis of Huntington's disease during the study. 37 of 40 cross-sectional and longitudinal clinical and imaging measures were significant predictors of motor diagnosis beyond CAG repeat length and age. The strongest predictors were in the motor, imaging, and cognitive domains: an increase of one SD in total motor score (motor domain) increased the risk of a motor diagnosis by 3·07 times (95% CI 2·26-4·16), a reduction of one SD in putamen volume (imaging domain) increased risk by 3·32 times (2·37-4·65), and a reduction of one SD in Stroop word score (cognitive domain) increased risk by 2·32 times (1·88-2·87). Prediction of diagnosis of Huntington's disease can be improved beyond that obtained by CAG repeat length and age alone. Such knowledge about potential predictors of manifest Huntington's disease should inform discussions about guidelines for diagnosis, prognosis, and counselling, and might be useful in guiding the selection of participants and outcome measures for clinical trials. US National Institutes of Health, US National Institute of Neurological Disorders and Stroke, and CHDI Foundation.


Syphilis in the Americas: a protocol for a systematic review of syphilis prevalence and incidence in four high-risk groups, 1980-2016.
Syphilis infection has recently resurfaced as a significant public health problem. Although there has been a tremendous amount of research on the epidemiology of syphilis, there has been limited work done to synthesize the extensive body of research and systematically estimate patterns of disease within high-risk groups in the Americas. The purpose of this systematic review and meta-analysis is to (1) summarize recent patterns of syphilis infection in North and South America among four high-risk groups (MSM, transgender women, sex workers, and incarcerated individuals) from 1980 to 2016, (2) identify and differentiate regional geographic epidemiologic characteristics, and (3) compare the epidemics of the economically developed countries of North America from the developing countries and public health systems of Latin America and the Caribbean. Primary studies reporting syphilis prevalence and/or incidence in at least one of the four high-risk groups will be identified from Medline/PubMed, Embase, Lilacs, SciELO, The Cochrane Library, Web of Science, Scopus, ProQuest, CINAHL, Clase, and Periódica, as well as "gray" literature sources (conference abstracts, country reports, etc.). Studies published from 1980 through 2016 will be included. Data will be extracted from studies meeting inclusion and exclusion criteria and a random effects meta-analysis of prevalence and incidence estimates will be conducted. Heterogeneity, risk of bias, and publication bias will be assessed. Pooled prevalence and incidence estimates will be calculated for comparisons based on geographic region, risk factors, and time period. Our systematic review and meta-analysis aims to contribute to an improved understanding of global epidemiologic patterns of syphilis infection in most-at-risk populations. Through systematic classification of the existing literature, and comparison of disease patterns across regional, temporal and socio-behavioral differences, we hope to improve public health surveillance and improve efforts to control the spread of disease across the Americas. PROSPERO CRD42016047306.


Fingerprint-based in silico models for the prediction of P-glycoprotein substrates and inhibitors.
P-Glycoprotein (P-gp, ABCB1) plays a significant role in determining the ADMET properties of drugs and drug candidates. Substrates of P-gp are not only subject to multidrug resistance (MDR) in tumor therapy, they are also associated with poor pharmacokinetic profiles. In contrast, inhibitors of P-gp have been advocated as modulators of MDR. However, due to the polyspecificity of P-gp, knowledge on the molecular basis of ligand-transporter interaction is still poor, which renders the prediction of whether a compound is a P-gp substrate/non-substrate or an inhibitor/non-inhibitor quite challenging. In the present investigation, we used a set of fingerprints representing the presence/absence of various functional groups for machine learning based classification of a set of 484 substrates/non-substrates and a set of 1935 inhibitors/non-inhibitors. Best models were obtained using a combination of a wrapper subset evaluator (WSE) with random forest (RF), kappa nearest neighbor (kNN) and support vector machine (SVM), showing accuracies >70%. Best P-gp substrate models were further validated with three sets of external P-gp substrate sources, which include Drug Bank (n = 134), TP Search (n = 90) and a set compiled from literature (n = 76). Association rule analysis explores the various structural feature requirements for P-gp substrates and inhibitors.


Effects of extreme prematurity and kindergarten neuropsychological skills on early academic progress.
The study was designed to investigate the effect of extreme prematurity on growth in academic achievement across the early school years and the validity of kindergarten neuropsychological skills as predictors of achievement. A 2001-2003 birth cohort of 145 extremely preterm/extremely low birth weight (EPT/ELBW) children from a single medical center, along with 111 normal birth weight (NBW) classmate controls, were recruited during their first year in kindergarten and followed annually across the next 2 years in school. Mixed model analysis was conducted to compare the groups on growth in achievement across years and examine kindergarten neuropsychological skills as predictors of growth. The EPT/ELBW group scored significantly below NBW controls on all achievement tests across years and had higher rates of special education placement and grade repetition. Despite limited catch-up of the EPT/ELBW group to the NBW controls in spelling, group differences were generally stable. Differences in spelling and mathematics achievement remained significant when controlling for global intelligence or excluding children who had intellectual or neurosensory impairments or repeated a grade. Higher scores on kindergarten tests of multiple neuropsychological ability domains predicted higher achievement levels and steeper growth in achievement. The findings document persistent academic weaknesses in EPT/ELBW children across the early school years. Results point to the need for preschool interventions to enhance academic readiness and suggest that neuropsychological skills assessed in kindergarten are useful in identifying individual differences in early learning progress. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Liquid state DNP for water accessibility measurements on spin-labeled membrane proteins at physiological temperatures.
We demonstrate the application of continuous wave dynamic nuclear polarization (DNP) at 0.35 T for site-specific water accessibility studies on spin-labeled membrane proteins at concentrations in the 10-100 μM range. The DNP effects at such low concentrations are weak and the experimentally achievable dynamic nuclear polarizations can be below the equilibrium polarization. This sensitivity problem is solved with an optimized home-built DNP probe head consisting of a dielectric microwave resonator and a saddle coil as close as possible to the sample. The performance of the probe head is demonstrated with both a modified pulsed EPR spectrometer and a dedicated CW EPR spectrometer equipped with a commercial NMR console. In comparison to a commercial pulsed ENDOR resonator, the home-built resonator has an FID detection sensitivity improvement of 2.15 and an electron spin excitation field improvement of 1.2. The reproducibility of the DNP results is tested on the water soluble maltose binding protein MalE of the ABC maltose importer, where we determine a net standard deviation of 9% in the primary DNP data in the concentration range between 10 and 100 μM. DNP parameters are measured in a spin-labeled membrane protein, namely the vitamin B(12) importer BtuCD in both detergent-solubilized and reconstituted states. The data obtained in different nucleotide states in the presence and absence of binding protein BtuF reveal the applicability of this technique to qualitatively extract water accessibility changes between different conformations by the ratio of primary DNP parameters ϵ. The ϵ-ratio unveils the physiologically relevant transmembrane communication in the transporter in terms of changes in water accessibility at the cytoplasmic gate of the protein induced by both BtuF binding at the periplasmic region of the transporter and ATP binding at the cytoplasmic nucleotide binding domains.


Screening for prostate cancer: an update.
To review evidence regarding the potential introduction of prostate cancer screening programmes and highlight issues pertinent to the management of screen-detected prostate cancer. Screening for prostate cancer is a controversial health care issue in general and urological practice. A PubMed database search was performed, followed by a systematic review of the literature, to examine the evidence base underlying prostate cancer screening. A prostate cancer screening programme should satisfy several key postulates prior to its introduction. To date, several of these postulates have not been satisfied, and the evidence available for prostate cancer screening is currently insufficient to warrant its introduction as a public health policy. The natural history of screen-detected prostate cancer remains poorly understood, and recent evidence suggests that a screening programme may detect a large number of men with indolent disease who may be subsequently overtreated. Several randomised clinical trials are currently in progress and it is hoped that they will provide robust evidence to inform future practice. National systematic prostate cancer screening programmes outside randomised clinical trial settings have not been implemented to date owing to lack of robust evidence that such programmes would improve survival and/or quality of life in men with screen-detected disease. Forthcoming results of clinical trials and the application of appropriate risk stratification to prevent overtreatment of indolent prostate cancer are likely to change practice in coming years.


Characterization and validation of new tools for measuring site-specific cardiac troponin I phosphorylation.
Phosphorylation of cardiac troponin I is a well established mechanism by which cardiac contractility is modulated. However, there are a number of phosphorylation sites on TnI which contribute singly or in combination to influence cardiac function. Accordingly, methods for accurately measuring site-specific TnI phosphorylation are needed. Currently, two strategies are employed: mass spectrometry, which is costly, difficult and has a low throughput; and Western blotting using phospho-specific antibodies, which is limited by the availability of reagents. In this report, we describe a cohort of new site-specific TnI phosphoantibodies, generated against physiologically relevant phosphorylation sites, that are superior to the current commercially available antibodies: to phospho-serine 22/23 which shows a >5-fold phospho-specificity for phosphorylated TnI; to phospho-serine 43, which has >3-fold phospho-specificity for phosphorylated TnI; and phospho-serine 150 which has >2-fold phospho-specificity for phosphorylated TnI. These new antibodies demonstrated greater sensitivity and specificity for the phosphorylated TnI than the most widely used commercially available reagents. For example, at a protein load of 20 μg of total cardiac extract, a commercially available antibody recognized both phosphorylated and dephosphorylated TnI to the same degree. At the same protein load our phospho-serine 22/23 antibody exhibited no cross-reactivity with dephosphorylated TnI. These new tools should allow a more accurate assessment and a better understanding of the role of TnI phosphorylation in the response of the heart to pathologic stress.


Application of central composite design and artificial neural network in modeling of reactive blue 21 dye removal by photo-ozonation process.
The present study deals with use of central composite design (CCD) and artificial neural network (ANN) in modeling and optimization of reactive blue 21 (RB21) removal from aqueous media under photo-ozonation process. Four effective operational parameters (including: initial concentration of RB21, O(3) concentration, UV light intensity and reaction time) were chosen and the experiments were designed by CCD based on response surface methodology (RSM). The obtained results from the CCD model were used in modeling the process by ANN. Under optimum condition (O(3) concentration of 3.95 mg L(-1), UV intensity of 20.5 W m(-2), reaction time of 7.77 min and initial dye concentration of 40.21 mg L(-1)), RB21 removal efficiency reached to up 98.88%. A topology of ANN with a three-layer consisting of four input neurons, 14 hidden neurons and one output neuron was designed. The relative significance of each major factor was calculated based on the connection weights of the ANN model. Dye and ozone concentrations were the most important variables in the photo-ozonation of RB21, followed by reaction time and UV light intensity. The comparison of predicted values by CCD and ANN with experimental results showed that both methods were highly efficient in the modeling of the process.


Myositis or dystrophy? Traps and pitfalls.
Some hereditary myopathies can mimic acquired myositis especially when they first present in adulthood with a limb-girdle distribution of weakness. Although inherited myopathies are generally painless and progress very slowly there are exceptions, which can further add to diagnostic confusion. The diagnosis is made even more difficult when inflammatory infiltrates are present on muscle biopsy. This is common in certain dystrophies in particular e.g. dysferlinopathies or facioscapulohumeral dystrophy. On the other hand, acquired (and treatable with immunosuppressants) necrotizing myopathies with anti-SRP antibodies can be very slowly progressive, with clinical and pathological features compatible with limb girdle dystrophies. These two situations can lead to either inappropriate immunosuppressant treatment in a patient with dystrophy, having mistaken it for an acquired inflammatory myopathy, or to therapeutic abstention in a patient with a treatable acquired myopathy thinking that it was a dystrophy. Pointers helping to distinguish between these two traps are here reviewed.


Formation of ZnO Micro-Flowers Prepared via Solution Process and their Antibacterial Activity.
This paper presents the fabrication and characterization of zinc oxide micro-flowers and their antibacterial activity. The micro-flowers of zinc oxide composed of hexagonal nanorods have been prepared via solution process using precursor zinc acetate di-hydrate and sodium hydroxide in 3 h of refluxing time at ~90°C. The antibacterial activities of grown micro-flowers were investigated against four pathogenic bacteria namely S. aureus, E. coli, S. typhimurium and K. pneumoniae by taking five different concentrations (5-45 μg/ml) of ZnO micro-flowers (ZnO-MFs). Our investigation reveals that at lowest concentration of ZnO-MFs solution inhibiting the growth of microbial strain which was found to be 5 μg/ml for all the tested pathogens. Additionally, on the basis of morphological and chemical observations, a chemical reaction mechanism of ZnO-MFs composed of hexagonal nanorods was also proposed.


A cellular function can enhance gene expression and plating efficiency of a mutant defective in the gene for ICP0, a transactivating protein of herpes simplex virus type 1.
ICP0 transactivates herpes simplex virus type 1 genes of all classes as well as a number of heterologous viral and cellular genes, yet it is not essential for virus replication in vitro or in vivo. Stocks of ICP0 deletion mutants, however, exhibit significantly lower plating efficiencies on standard 24-h-old Vero cell monolayers than do stocks of wild-type virus. In an attempt to determine whether the growth status of cells in the monolayer affects the ability of ICP0 mutants to initiate plaque formation, the plating efficiencies and abilities of an ICP0 null mutant (7134) and of wild-type virus (KOS) to express selected viral proteins were determined on Vero cell monolayers whose growth had been arrested either by contact inhibition-trypsinization or by isoleucine deprivation and had then been released from growth arrest. The proportion of cells cycling synchronously after release from growth arrest was assessed by flow cytometry. The results of these studies indicate that the plating efficiency of 7134 was greatest on Vero cell monolayers 8 h after release from growth arrest induced by either treatment. Monolayers of both types released from growth arrest at other times supported 7134 plaque formation less efficiently. In contrast, the plating efficiency of KOS was nearly equal on monolayers at all times after release from growth arrest. Notably, both KOS and 7134 were equally efficient in entering cells and inducing expression of the immediate-early protein ICP4 in either 8- or 24-h monolayers. Relative to KOS, however, 7134 was significantly impaired in the expression of selected early and late genes in cells at 24 h postrelease. When the plating efficiencies of 7134 and KOS were examined in 0-28 cells (Vero cells that are stably transformed with the ICP0 gene) whose growth had been arrested and then released, no differences in the plating efficiencies of the two viruses as a function of growth status were noted. These findings suggest that a cellular function expressed maximally in cells 8 h after release from growth arrest can substitute operationally for ICP0 to enhance plaque formation and viral gene expression by 7134. They further suggest that one role of ICP0 in viral infection is to facilitate virus replication in cells that do not express this function.


Hyperbaric oxygenation modulates vascular reactivity to angiotensin-(1-7) in diabetic rats: potential role of epoxyeicosatrienoic acids.
Previously, a facilitating effect of hyperbaric oxygenation (HBO₂) on aortic ring responses to angiotensin-(1-7) in healthy rats was reported, with epoxyeicosatrienoic acids (EETs) possibly playing an important role. The aim of this study was to assess whether HBO₂ exerts similar effects in diabetic rats and to further explore the role of specific cytochrome P450 (CYP) enzymes in changes induced by HBO₂. Aortic relaxation to angiotensin-(1-7) was significantly higher in HBO₂ diabetic rats compared to control diabetic rats, while HBO₂ had no effect on angiotensin II contraction. N-methylsulphonyl-6-(2-propargyloxyphenyl/hexanamide inhibited the facilitation of angiotensin-(1-7) responses in HBO₂ rats, suggesting an important role of EETs in this modulation. mRNA expression of CYP2J3 and protein expression of CYP2C11 were significantly upregulated in HBO₂ diabetic rats, whereas CYP4A1, CYP4A2 and CYP4A3 mRNA and CYP2J3 protein expression was similar between groups. Mean arterial pressure, ferric reducing ability of plasma and Thiobarbituric Acid Reactive Substances levels and serum angiotensin-(1-7) concentrations were not significantly changed.


Comparison of Londrina activities of daily living protocol and Glittre ADL test on cardio-pulmonary response in patients with COPD: a cross-sectional study.
In COPD patients it is very important to assess the activities of daily living (ADL) due to an impairment of independence and quality of life. There is a lack of retrievable data regarding the cardio-pulmonary response to Londrina ADL protocol in patients with COPD. The aim of the present study was to assess the cardio-pulmonary response to Londrina ADL protocol in patients with COPD and to compare this with responses to the Glittre ADL test. This cross-sectional study was done on 30 COPD subjects. Each subject was taken to perform the Londrina ADL protocol, Glittre ADL test, twice each, on subsequent days. The Londrina ADL protocol comprises 5 activities representing ADL, involving upper limbs, lower limbs, and trunk movements. The Glittre ADL test consists of completing a circuit while carrying a weighted backpack (2.5 kg for women, 5.0 kg for men). The better value of the two was taken into consideration. For the Londrina ADL protocol and Glittre ADL test the outcome of primary interest was time and for the six-minute walk test was the distance walked. The secondary outcomes for all tests were heart rate, systolic blood pressure, diastolic blood pressure, respiratory rate, saturation of oxygen in blood and dyspnea. The COPD subjects of age group 63.27±11.07 years took 5.94±0.36 min to complete trial 2 of the Londrina ADL protocol. Significant physiological increases in heart rate (p≤0.01), respiratory rate (p≤0.01), blood pressure (p≤0.01) and severity of dyspnea (p≤0.01) were observed, whereas saturation of oxygen in blood (p≤0.01) was reduced at the end of the Londrina ADL protocol and Glittre ADL test. There was a positive, non-significant correlation between the six-minute walk test distance and the Londrina ADL protocol time (r=0.236) (p=0.209). A positive, not significant correlation was observed between the Glittre ADL test (time) and the Londrina ADL protocol (time) (r=0.194) (p=0.304) and a negative but not significant correlation between the six-minute walk test (distance) and the Glittre ADL test (time) (r= -0.184) (p=0.330). The Londrina ADL protocol can be used as an assessment tool for the evaluation of functional performance and activities of daily living in COPD along with other test protocols in pulmonary rehabilitation.


Dr Google and the consumer: a qualitative study exploring the navigational needs and online health information-seeking behaviors of consumers with chronic health conditions.
The abundance of health information available online provides consumers with greater access to information pertinent to the management of health conditions. This is particularly important given an increasing drive for consumer-focused health care models globally, especially in the management of chronic health conditions, and in recognition of challenges faced by lay consumers with finding, understanding, and acting on health information sourced online. There is a paucity of literature exploring the navigational needs of consumers with regards to accessing online health information. Further, existing interventions appear to be didactic in nature, and it is unclear whether such interventions appeal to consumers' needs. Our goal was to explore the navigational needs of consumers with chronic health conditions in finding online health information within the broader context of consumers' online health information-seeking behaviors. Potential barriers to online navigation were also identified. Semistructured interviews were conducted with adult consumers who reported using the Internet for health information and had at least one chronic health condition. Participants were recruited from nine metropolitan community pharmacies within Western Australia, as well as through various media channels. Interviews were audio-recorded, transcribed verbatim, and then imported into QSR NVivo 10. Two established approaches to thematic analysis were adopted. First, a data-driven approach was used to minimize potential bias in analysis and improve construct and criterion validity. A theory-driven approach was subsequently used to confirm themes identified by the former approach and to ensure identified themes were relevant to the objectives. Two levels of analysis were conducted for both data-driven and theory-driven approaches: manifest-level analysis, whereby face-value themes were identified, and latent-level analysis, whereby underlying concepts were identified. We conducted 17 interviews, with data saturation achieved by the 14th interview. While we identified a broad range of online health information-seeking behaviors, most related to information discussed during consumer-health professional consultations such as looking for information about medication side effects. The barriers we identified included intrinsic barriers, such as limited eHealth literacy, and extrinsic barriers, such as the inconsistency of information between different online sources. The navigational needs of our participants were extrinsic in nature and included health professionals directing consumers to appropriate online resources and better filtering of online health information. Our participants' online health information-seeking behaviors, reported barriers, and navigational needs were underpinned by the themes of trust, patient activation, and relevance. This study suggests that existing interventions aimed to assist consumers with navigating online health information may not be what consumers want or perceive they need. eHealth literacy and patient activation appear to be prevalent concepts in the context of consumers' online health information-seeking behaviors. Furthermore, the role for health professionals in guiding consumers to quality online health information is highlighted.


Interprofessional relationships and communication in primary palliative care: impact of the Gold Standards Framework.
High quality end-of-life care in the community is achieved with effective multidisciplinary teamwork, interprofessional communication between GPs and district nurses, and early referral of patients to district nurses. These aspects of palliative care are highlighted in the Gold Standards Framework, a programme recently established in UK primary care. To investigate the extent to which the framework influences interprofessional relationships and communication, and to compare GPs' and nurses' experiences. Qualitative interview case study. Fifteen participating practices from three primary care trusts in England. Thirty-eight semi-structured interviews were undertaken with GPs, district nurses, Macmillan nurses, and framework facilitators. Adoption of the framework often resulted in earlier referral of palliative care patients to district nurses. Multidisciplinary team meetings enabled communication for sharing knowledge, discussing management problems, and keeping colleagues informed; however, arranging and maintaining such meetings was often problematic. Nurses particularly valued formal meetings while GPs generally preferred informal ad hoc dialogue. GPs largely maintained control of the mode of multidisciplinary working. The best functioning teams used a mixture of formal and informal meetings with a relatively non-hierarchical working style. Implementing the framework enabled processes of communication associated with high quality palliative care in general practice, but there was marked variation in how this worked in individual teams. In general, hierarchical doctor-nurse relationships persisted.


In Search of 'Omics'-Based Biomarkers to Predict Risk of Frailty and Its Consequences in Older Individuals: The FRAILOMIC Initiative.
An increase in the number of older people experiencing disability and dependence is a critical aspect of the demographic change that will emerge within Europe due to the rise in life expectancy. In this scenario, prevention of these conditions is crucial for the well-being of older citizens and for the sustainability of our healthcare systems. Thus, the diagnosis and management of conditions like frailty, which identifies the people at the highest risk for developing those adverse outcomes, is of critical relevance. Currently, assessment of frailty relies primarily on measuring functional parameters, which have limited clinical utility. In this viewpoint article, we describe the FRAILOMIC Initiative, an international, large-scale, multi-endpoint, community- and clinic-based research study funded by the European Commission. The aim of the study is to develop validated measures, comprising both classic and 'omics-based' laboratory biomarkers, which can predict the risk of frailty, improve the accuracy of its diagnosis in clinical practice and provide a prognostic forecast on the evolution from frailty to disability. The initiative includes eight established cohorts of older adults, encompassing >75,000 subjects, most of whom (∼70%) are aged >65 years. Data on function, nutritional status and exercise habits have been collected, and cardiovascular health has been evaluated at baseline. Subjects will be stratified as 'non-frail' or 'frail' using Fried's definition, all adverse outcomes of interest will be recorded and differentially expressed biomarkers associated with the risk of frailty will be identified. Genomic, proteomic and transcriptomic investigations will be carried out using array-based systems. As circulating microRNAs in plasma have been identified in the context of senescence, ageing and age-associated diseases, a miRNome-wide analysis will also be undertaken to identify a miRNA-based signature of frailty. Blood concentrations of secreted proteins known to be upregulated significantly in senescent endothelial cells and other hypothesis-driven biomarkers will be measured using ELISAs. The FRAILOMIC Initiative aims to issue a series of interim scientific reports as key results emerge. Ultimately, it is hoped that this study will contribute to the development of new clinical tools, which may help individuals to enjoy an old age that is healthier and free from disability.


The endogenous cannabinoid, anandamide, induces cell death in colorectal carcinoma cells: a possible role for cyclooxygenase 2.
Cyclooxygenase 2 (COX-2) is upregulated in most colorectal cancers and is responsible for metabolism of the endogenous cannabinoid, anandamide, into prostaglandin-ethanolamides (PG-EAs). The aims of this study were to determine whether anandamide and PG-EAs induce cell death in colorectal carcinoma (CRC) cells, and whether high levels of COX-2 in CRC cells could be utilised for their specific targeting for cell death by anandamide. We determined the effect of anandamide on human CRC cell growth by measuring cell growth and cell death, whether this was dependent on COX-2 protein expression or enzyme activity, and the potential involvement of PG-EAs in induction of cell death. Anandamide inhibited the growth of CRC cell lines HT29 and HCA7/C29 (moderate and high COX-2 expressors, respectively) but had little effect on the very low COX-2 expressing CRC cell line, SW480. Induction of cell death in HT29 and HCA7/C29 cell lines was partially rescued by the COX-2 selective inhibitor NS398. Cell death induced by anandamide was neither apoptosis nor necrosis. Furthermore, inhibition of fatty acid amide hydrolase potentiated the non-apoptotic cell death, indicating that anandamide induced cell death was mediated via metabolism of anandamide by COX-2, rather than its degradation into arachidonic acid and ethanolamine. Interestingly, both PGE2-EA and PGD2-EA induced classical apoptosis. These findings suggest anandamide may be a useful chemopreventive/therapeutic agent for colorectal cancer as it targets cells that are high expressors of COX-2, and may also be used in the eradication of tumour cells that have become resistant to apoptosis.


Novel genetic loci underlying human intracranial volume identified through genome-wide association.
Intracranial volume reflects the maximally attained brain size during development, and remains stable with loss of tissue in late life. It is highly heritable, but the underlying genes remain largely undetermined. In a genome-wide association study of 32,438 adults, we discovered five previously unknown loci for intracranial volume and confirmed two known signals. Four of the loci were also associated with adult human stature, but these remained associated with intracranial volume after adjusting for height. We found a high genetic correlation with child head circumference (ρgenetic = 0.748), which indicates a similar genetic background and allowed us to identify four additional loci through meta-analysis (Ncombined = 37,345). Variants for intracranial volume were also related to childhood and adult cognitive function, and Parkinson's disease, and were enriched near genes involved in growth pathways, including PI3K-AKT signaling. These findings identify the biological underpinnings of intracranial volume and their link to physiological and pathological traits.