Test-retest reliability of time-frequency measures of auditory steady-state responses in patients with schizophrenia and healthy controls.

BackgroundAuditory steady-state response (ASSR) paradigms have consistently demonstrated gamma band abnormalities in schizophrenia at a 40-Hz driving frequency with both electroencephalography (EEG) and magnetoencephalography (MEG). Various time-frequency measures have been used to assess the 40-Hz ASSR, including evoked power, single trial total power, phase-locking factor (PLF), and phase-locking angle (PLA). While both EEG and MEG studies have shown power and PLF ASSR measures to exhibit excellent test-retest reliability in healthy adults, the reliability of these measures in patients with schizophrenia has not been determined.MethodsASSRs were obtained by recording EEG data during presentation of repeated 20-Hz, 30-Hz and 40-Hz auditory click trains from nine schizophrenia patients (SZ) and nine healthy controls (HC) tested on two occasions. Similar ASSR data were collected from a separate group of 30 HC on two to three test occasions. A subset of these HC subjects had EEG recordings during two tasks, passively listening and actively attending to click train stimuli. Evoked power, total power, PLF, and PLA were calculated following Morlet wavelet time-frequency decomposition of EEG data and test-retest generalizability (G) coefficients were calculated for each ASSR condition, time-frequency measure, and subject group.ResultsG-coefficients ranged from good to excellent (> 0.6) for most 40-Hz time-frequency measures and participant groups, whereas 20-Hz G-coefficients were much more variable. Importantly, test-retest reliability was excellent for the various 40-Hz ASSR measures in SZ, similar to reliabilities in HC. Active attention to click train stimuli modestly reduced G-coefficients in HC relative to the passive listening condition.DiscussionThe excellent test-retest reliability of 40-Hz ASSR measures replicates previous EEG and MEG studies. PLA, a relatively new time-frequency measure, was shown for the first time to have excellent reliability, comparable to power and PLF measures. Excellent reliability of 40 Hz ASSR measures in SZ supports their use in clinical trials and longitudinal observational studies

Palladium und Kupfer-katalysierte Domino- und Multikomponentenreaktionen auf Basis der Kupplungs-Isomerisierungs-Reaktion

Die Kupplungs-Isomerisierungs-Reaktion (KIR), eine schnelle Sonogashira-Alkinylierung von elektronenziehenden Halogeniden und Propargylalkoholen gefolgt von einer basenkatalysierten Carbinol-Allenol-Enon-Isomerisierung, stellt einen milden und effizienten Zugang zu Chalkonen dar. Innerhalb der vorliegenden Arbeit konnten neue Dominoreaktionen entwickelt werden, die methodische Erweiterungen der KIR darstellen. Es ist gelungen die zwingend notwendige Akzeptorfunktionalität in situ mittels einer intramolekularen Carbopalladierungsreaktion zu erzeugen. Terminale Acetylene reagieren mit N-Iodphenylalkinylamiden in einer Carbopalladierungs-Sonogashira-Alkinylierungs-Dominosequenz zu Inylidendihydroindolonen; der Alkinsäureiodphenylester konnte mit 4 Anisylpropargylalkohol in einer Insertions-Kupplungs-Isomerisierungs-Reaktion zum Oxobutenylidenbenzofuranon umgesetzt werden. Entgegen dem Postulat, dass intramolekulare 5-exo-dig-Cyclisierungsreaktionen streng syn-stereospezifisch verlaufen, wurden die Inylidendihydroindolone als E/Z-Doppelbindungsisomere und das Oxobutenylidenbenzofuranon anti-stereoselektiv als Z Isomer erhalten. Neben der Einbindung eines metallorganischen Insertionsereignisses konnte zudem die irreversible Allenol-Enon-Tautomerie durch die Inkorporation von Propargylethern anstelle der entsprechenden Alkohole erfolgreich unterbunden werden. Das bislang elusive, hoch reaktive Allenintermediat kann jetzt in einer kinetisch und thermodynamisch kontrollierten intramolekularen Abfangreaktion adressiert werden. Die Insertions-KI-Diels-Alder-Dominoreaktion der Akzeptoren bzw. mit Propargylallylethern liefert die bislang unbekannte, stark fluoreszierende Substanzklasse der Spirobenzofuranone und Spiroindolone in moderaten bis guten Ausbeuten. DFT- und MP2-Rechnungen der konkurrierenden produktbildenden Schritte der intramolekularen Diels-Alder-Reaktion vs. der Claisen-Umlagerung des Vinylallenylallyletherintermediats zeigen sowohl eine kinetische als auch thermodynamische Präferenz der [4+2]-Cycloaddition. Mit der Erkenntnis dieser Molecular-Modeling Studien konnten die elektronische Natur und die sterischen Eigenschaften der Akzeptor- und Alkinkomponente so modifiziert werden, dass nun eine Claisen-Umlagerung als Folgereaktion das Intermediat adressiert. Die Kupplungs-Isomerisierungs-Claisen-Dominoreaktion von elektronenarmen Arylhalogeniden bzw. aromatischen Säurechloriden mit Propargyltritylethern zeigt eine bemerkenswerte Dichotomie im finalen Schritt und resultiert, als Folge selbst kleinster elektronisch diverser Substituenteneffekte, in der Bildung von Tricyclooctenen, Chalkone, 1H-Iso¬chro¬menen und Indanen. Die Sonogashira-Reaktion von terminalen Phenylacetylenen mit N-Iodphenylalkinylamiden bzw. Alkinsäureiodphenylestern liefert in einer Carbopalladierungs-Alkinylierungs-Reaktion die entsprechenden internen vinylogen Alkinoylderivate. Erfolgt die Umsetzung der in situ generierten Alkinoylester bzw. -amide mit primären oder sekundären Aminen im Sinne einer Aza-Michael-Addition, ist ein eleganter Zugang zu vinylogen Enaminocarbonsäureestern und -amiden geschaffen. Unter Verwendung der hier vorgestellten Insertions-Sonogashira-Aminovinylierungs-Dreikomponentenreaktion können im Ein-Topf-Verfahren maßgeschneiderte orangefarben bis rot fluoreszierende Push-Pull-Chromophore und in guten bis exzellenten Ausbeuten erhalten werden

Should patients with abnormal liver function tests in primary care be tested for chronic viral hepatitis: cost minimisation analysis based on a comprehensively tested cohort

Background Liver function tests (LFTs) are ordered in large numbers in primary care, and the Birmingham and Lambeth Liver Evaluation Testing Strategies (BALLETS) study was set up to assess their usefulness in patients with no pre-existing or self-evident liver disease. All patients were tested for chronic viral hepatitis thereby providing an opportunity to compare various strategies for detection of this serious treatable disease. Methods This study uses data from the BALLETS cohort to compare various testing strategies for viral hepatitis in patients who had received an abnormal LFT result. The aim was to inform a strategy for identification of patients with chronic viral hepatitis. We used a cost-minimisation analysis to define a base case and then calculated the incremental cost per case detected to inform a strategy that could guide testing for chronic viral hepatitis. Results Of the 1,236 study patients with an abnormal LFT, 13 had chronic viral hepatitis (nine hepatitis B and four hepatitis C). The strategy advocated by the current guidelines (repeating the LFT with a view to testing for specific disease if it remained abnormal) was less efficient (more expensive per case detected) than a simple policy of testing all patients for viral hepatitis without repeating LFTs. A more selective strategy of viral testing all patients for viral hepatitis if they were born in countries where viral hepatitis was prevalent provided high efficiency with little loss of sensitivity. A notably high alanine aminotransferase (ALT) level (greater than twice the upper limit of normal) on the initial ALT test had high predictive value, but was insensitive, missing half the cases of viral infection. Conclusions Based on this analysis and on widely accepted clinical principles, a "fast and frugal" heuristic was produced to guide general practitioners with respect to diagnosing cases of viral hepatitis in asymptomatic patients with abnormal LFTs. It recommends testing all patients where a clear clinical indication of infection is present (e.g. evidence of intravenous drug use), followed by testing all patients who originated from countries where viral hepatitis is prevalent, and finally testing those who have a notably raised ALT level (more than twice the upper limit of normal). Patients not picked up by this efficient algorithm had a risk of chronic viral hepatitis that is lower than the general population

Polyclonal mucosa-associated invariant T cells have unique innate functions in bacterial infection

Mucosa-associated invariant T (MAIT) cells are a unique population of αβ T cells in mammals that reside preferentially in mucosal tissues and express an invariant Vα paired with limited Vβ T-cell receptor (TCR) chains. Furthermore, MAIT cell development is dependent upon the expression of the evolutionarily conserved major histocompatibility complex (MHC) class Ib molecule MR1. Using in vitro assays, recent studies have shown that mouse and human MAIT cells are activated by antigen-presenting cells (APCs) infected with diverse microbes, including numerous bacterial strains and yeasts, but not viral pathogens. However, whether MAIT cells play an important, and perhaps unique, role in controlling microbial infection has remained unclear. To probe MAIT cell function, we show here that purified polyclonal MAIT cells potently inhibit intracellular bacterial growth of Mycobacterium bovis BCG in macrophages (MΦ) in coculture assays, and this inhibitory activity was dependent upon MAIT cell selection by MR1, secretion of gamma interferon (IFN-γ), and an innate interleukin 12 (IL-12) signal from infected MΦ. Surprisingly, however, the cognate recognition of MR1 by MAIT cells on the infected MΦ was found to play only a minor role in MAIT cell effector function. We also report that MAIT cell-deficient mice had higher bacterial loads at early times after infection compared to wild-type (WT) mice, demonstrating that MAIT cells play a unique role among innate lymphocytes in protective immunity against bacterial infection

Evaluation and Recommendations on Good Clinical Laboratory Practice Guidelines for Phase I–III Clinical Trials

Marcella Sarzotti-Kelsoe and colleagues harmonize various approaches to Good Clinical Laboratory Practice for clinical trials into a single set of recommendations

rAAV-compatible MiniPromoters for restricted expression in the brain and eye

Abstract Background Small promoters that recapitulate endogenous gene expression patterns are important for basic, preclinical, and now clinical research. Recently, there has been a promising revival of gene therapy for diseases with unmet therapeutic needs. To date, most gene therapies have used viral-based ubiquitous promoters–however, promoters that restrict expression to target cells will minimize off-target side effects, broaden the palette of deliverable therapeutics, and thereby improve safety and efficacy. Here, we take steps towards filling the need for such promoters by developing a high-throughput pipeline that goes from genome-based bioinformatic design to rapid testing in vivo. Methods For much of this work, therapeutically interesting Pleiades MiniPromoters (MiniPs; ~4 kb human DNA regulatory elements), previously tested in knock-in mice, were “cut down” to ~2.5 kb and tested in recombinant adeno-associated virus (rAAV), the virus of choice for gene therapy of the central nervous system. To evaluate our methods, we generated 29 experimental rAAV2/9 viruses carrying 19 different MiniPs, which were injected intravenously into neonatal mice to allow broad unbiased distribution, and characterized in neural tissues by X-gal immunohistochemistry for icre, or immunofluorescent detection of GFP. Results The data showed that 16 of the 19 (84 %) MiniPs recapitulated the expression pattern of their design source. This included expression of: Ple67 in brain raphe nuclei; Ple155 in Purkinje cells of the cerebellum, and retinal bipolar ON cells; Ple261 in endothelial cells of brain blood vessels; and Ple264 in retinal Müller glia. Conclusions Overall, the methodology and MiniPs presented here represent important advances for basic and preclinical research, and may enable a paradigm shift in gene therapy

eIMRT: a web platform for the verification and optimization of radiation treatment plans

The eIMRT platform is a remote distributed computing tool that provides users with Internet access to three different services: Monte Carlo optimization of treatment plans, CRT & IMRT treatment optimization, and a database of relevant radiation treatments/clinical cases. These services are accessible through a user-friendly and platform independent web page. Its flexible and scalable design focuses on providing the final users with services rather than a collection of software pieces. All input and output data (CT, contours, treatment plans and dose distributions) are handled using the DICOM format. The design, implementation, and support of the verification and optimization algorithms are hidden to the user. This allows a unified, robust handling of the software and hardware that enables these computation-intensive services. The eIMRT platform is currently hosted by the Galician Supercomputing Center (CESGA) and may be accessible upon request (there is a demo version at http://eimrt.cesga.es:8080/ eIMRT2/demo; request access in http://eimrt.cesga.es/signup.html). This paper describes all aspects of the eIMRT algorithms in depth, its user interface, and its services. Due to the flexible design of the platform, it has numerous applications including the intercenter comparison of treatment planning, the quality assurance of radiation treatments, the design and implementation of new approaches to certain types of treatments, and the sharing of information on radiation treatment techniques. In addition, the web platform and software tools developed for treatment verification and optimization have a modular design that allows the user to extend them with new algorithms. This software is not a commercial product. It is the result of the collaborative effort of different public research institutions and is planned to be distributed as an open source project. In this way, it will be available to any user; new releases will be generated with the new implemented codes or upgradesThis work was financed by Xunta de Galicia of Spain through grant PGIDT05SIN00101CT and by the European Community through the BeInGrid projectS

A review of the ecological value of Cusuco National Park an urgent call forconservation action in a highly threatened Mesoamerican cloud forest

Cloud forests are amongst the most biologically unique, yet threatened, ecosystems in Mesoamerica. We summarize the ecological value and conservation status of a well-studied cloud forest site: Cusuco National Park (CNP), a 23,440 ha protected area in the Merendón mountains, northwest Honduras. We show CNP to have exceptional biodiversity; of 966 taxa identified to a species-level to date, 362 (37.5%) are Mesoamerican endemics, 67 are red-listed by the IUCN, and at least 49 are micro-endemics known only from the Merendón range. CNP also provides key ecosystem services including provision of drinking water and downstream flood mitigation, as well as carbon sequestration, with an estimated stock of 3.5 million megagrams of carbon in 2000. Despite its ecological importance, CNP faces multiple environmental threats and associated stresses, including deforestation (1,759 ha since 2000 equating to 7% of total forest area), poaching (7% loss of mammal relative abundance per year), amphibian declines due to chytridiomycosis (70% of species threatened or near-threatened), and climate change (a mean 2.6 °C increase in temperature and 112 mm decrease in rainfall by 2100). Despite conservation actions, including community ranger patrols, captive-breeding programmes, and ecotourism initiatives, environmental degradation of CNP continues. Further action is urgently required, including reinforcement and expansion of ranger programmes, greater stakeholder engagement, community education programmes, development of alternative livelihood projects, and legislative enforcement and prosecution. Without a thorough and rapid response to understand and mitigate illegal activities, the extirpation and extinction of species and the loss of vital ecosystem services are inevitable in the coming decades

Prediction of Co-Receptor Usage of HIV-1 from Genotype

Human Immunodeficiency Virus 1 uses for entry into host cells a receptor (CD4) and one of two co-receptors (CCR5 or CXCR4). Recently, a new class of antiretroviral drugs has entered clinical practice that specifically bind to the co-receptor CCR5, and thus inhibit virus entry. Accurate prediction of the co-receptor used by the virus in the patient is important as it allows for personalized selection of effective drugs and prognosis of disease progression. We have investigated whether it is possible to predict co-receptor usage accurately by analyzing the amino acid sequence of the main determinant of co-receptor usage, i.e., the third variable loop V3 of the gp120 protein. We developed a two-level machine learning approach that in the first level considers two different properties important for protein-protein binding derived from structural models of V3 and V3 sequences. The second level combines the two predictions of the first level. The two-level method predicts usage of CXCR4 co-receptor for new V3 sequences within seconds, with an area under the ROC curve of 0.937±0.004. Moreover, it is relatively robust against insertions and deletions, which frequently occur in V3. The approach could help clinicians to find optimal personalized treatments, and it offers new insights into the molecular basis of co-receptor usage. For instance, it quantifies the importance for co-receptor usage of a pocket that probably is responsible for binding sulfated tyrosine