Infrared imaging of high density protein arrays.
We propose in this paper that protein microarrays could be analysed by infrared imaging in place of enzymatic or fluorescence labelling. This label-free method reports simultaneously a large series of data on the spotted sample (protein secondary structure, phosphorylation, glycosylation, presence of impurities, etc.). In the present work, 100 μm protein spots each containing about 100 pg protein were deposited to form high density regular arrays. Using arrays of infrared detectors, high resolution images could be obtained where each pixel of the image is in fact a full infrared spectrum. With microarrays, hundreds of experimental conditions can be tested easily and quickly, with no further labelling or chemistry of any kind. We describe how the noise present in the infrared spectra can be split into image noise and detector noise. We also detail how both types of noise can be most conveniently dealt with to generate very high quality spectra of less than 100 pg protein. Finally, the results suggest that the protein secondary structure is preserved during microarray building.


Experience with exchange and archiving of raw data: comparison of data from two diffractometers and four software packages on a series of lysozyme crystals.
The International Union of Crystallography has for many years been advocating archiving of raw data to accompany structural papers. Recently, it initiated the formation of the Diffraction Data Deposition Working Group with the aim of developing standards for the representation of these data. A means of studying this issue is to submit exemplar publications with associated raw data and metadata. A recent study on the effects of dimethyl sulfoxide on the binding of cisplatin and carboplatin to histidine in 11 different lysozyme crystals from two diffractometers led to an investigation of the possible effects of the equipment and X-ray diffraction data processing software on the calculated occupancies and B factors of the bound Pt compounds. 35.3 Gb of data were transferred from Manchester to Utrecht to be processed with EVAL. A systematic comparison shows that the largest differences in the occupancies and B factors of the bound Pt compounds are due to the software, but the equipment also has a noticeable effect. A detailed description of and discussion on the availability of metadata is given. By making these raw diffraction data sets available via a local depository, it is possible for the diffraction community to make their own evaluation as they may wish.


Gonococcal opacity protein promotes bacterial entry-associated rearrangements of the epithelial cell actin cytoskeleton.
Neisseria gonorrhoeae enters cultured human mucosal cells following binding of a distinct gonococcal opacity (Opa) outer membrane protein to cell surface proteoglycan receptors. We examined the route of internalization that is activated by Opa-expressing gonococci (strain VP1). Microscopy of infected Chang epithelial cells showed that gonococcal uptake was insensitive to monodansylcadaverine (150 microM), which interferes with clathrin-mediated endocytosis. Similarly, indirect immunofluorescence staining for clathrin in infected cells showed distribution of cellular clathrin unaltered from the distribution in noninfected cells. The microtubule inhibitors colchicine (50 microM) and nocodazole (20 microM) but not the microtubule-stabilizing agent taxol (10 microM) caused a moderate (30 to 50%) reduction in gonococcal entry without affecting bacterial adherence. The most dramatic effects were obtained with the microfilament-disrupting agent cytochalasin D (3 microM), which totally blocked bacterial entry into the cells. Double immunofluorescence staining of gonococci and actin filaments in infected cells demonstrated bacterium-associated accumulations of F-actin as an early signal of bacterial entry. The recruitment of F-actin was transient and disappeared once the bacteria were inside the cells. Cytochalasin D disrupted the actin cytoskeleton architecture but did not prevent the recruitment of F-actin by the bacteria. Adherent, noninvasive gonococcal Opa variants lacked the ability to mobilize F-actin. Recombinant Escherichia coli expressing the gonococcal invasion-promoting Opa of gonococcal strain MS11 (Opa50) adhered to the epithelial cells in an Opa-dependent fashion but was not internalized and did not recruit detectable amounts of F-actin. Coinfection with the E. coli recombinant strain and gonococci resulted in specific entry of the diplococci, despite the presence of large numbers of adherent E. coli cells. Together, our results indicate that Opa-mediated gonococcal entry into Chang cells resembles phagocytosis rather than macropinocytosis reported for Salmonella spp. and sequentially involves gonococcal adherence to the cell surface, Opa-dependent and cytochalasin-insensitive recruitment of F-actin, and cytochalasin D-sensitive bacterial internalization.


Importance of a Fully Anharmonic Treatment of Equilibrium Isotope Fractionation Properties of Dissolved Ionic Species As Evidenced by Li+(aq).
Equilibrium fractionation of stable isotopes is critically important in fields ranging from chemistry, including medicinal chemistry, electrochemistry, geochemistry, and nuclear chemistry, to environmental science. The dearth of reliable estimates of equilibrium fractionation factors, from experiment or from natural observations, has created a need for accurate computational approaches. Because isotope fractionation is a purely quantum mechanical phenomenon, exact calculation of fractionation factors is nontrivial. Consequently, a severe approximation is often made, in which it is assumed that the system can be decomposed into a set of independent harmonic oscillators. Reliance on this often crude approximation is one of the primary reasons that theoretical prediction of isotope fractionation has lagged behind experiment. A class of problems for which one might expect the harmonic approximation to perform most poorly is the isotopic fractionation between solid and solution phases. In order to illustrate the errors associated with the harmonic approximation, we have considered the fractionation of Li isotopes between aqueous solution and phyllosilicate minerals, where we find that the harmonic approximation overestimates isotope fractionation factors by as much as 30% at 25 °C. Lithium is a particularly interesting species to examine, as natural lithium isotope signatures provide information about hydrothermal processes, carbon cycle, and regulation of the Earth's climate by continental alteration. Further, separation of lithium isotopes is of growing interest in the nuclear industry due to a need for pure 6Li and 7Li isotopes. Moving beyond the harmonic approximation entails performing exact quantum calculations, which can be achieved using the Feynman path integral formulation of quantum statistical mechanics. In the path integral approach, a system of quantum particles is represented as a set of classical-like ring-polymer chains, whose interparticle interactions are determined by the rules of quantum mechanics. Because a classical isomorphism exists between the true quantum system and the system of ring-polymers, classical-like methods can be applied. Recent developments of efficient path integral approaches for the exact calculation of isotope fractionation now allow the case of the aforementioned dissolved Li fractionation properties to be studied in detail. Applying this technique, we find that the calculations yield results that are in good agreement with both experimental data and natural observations. Importantly, path integral methods, being fully atomistic, allow us to identify the origins of anharmonic effects and to make reliable predictions at temperatures that are experimentally inaccessible yet are, nevertheless, relevant for natural phenomena.


Well-defined set of exponents for a pair contact process with diffusion.
Recently it was suggested that a pair contact process with diffusion (PCPD) might represent an independent new universality class different from the directed percolation (DP) and the parity conservation (PC) class. The dynamics in the PCPD are usually controlled by two independent parameters. The critical exponents for the PCPD are known to have different values for varying values of the two independent parameters. However, once the diffusion and annihilation (or coagulation) rate in the PCPD is tuned in a way that the process without offspring production is exactly solvable, a well-defined set of the exponents for the PCPD is obtained. Then dynamics are controlled by only one independent parameter. The obtained critical exponents are different than those of DP and PC. The critical exponents satisfy the generalized hyperscaling relation within numerical errors.


Species invasions and the limits to restoration: learning from the New Zealand experience.
Species invasions impose key biotic thresholds limiting the success of ecological restoration projects. These thresholds may be difficult to reverse and will have long-term consequences for restoration because of invasion legacies such as extinctions; because most invasive species cannot be eliminated given current technology and resources; and because even when controlled to low levels, invasive species continue to exert substantial pressure on native biodiversity. Restoration outcomes in the face of biological invasions are likely to be novel and will require long-term resource commitment, as any letup in invasive species management will result in the loss of the conservation gains achieved.


Contractile and fatigue properties of thyrotoxic rat skeletal muscle.
The effects of thyrotoxicosis on the contractile properties and development of muscle fatigue in the slow soleus (SOL) and fast extensor digitorum longus (EDL) muscles were examined in rats given 3 mg of L-thyroxine and 1 mg of L-triiodothyronine per kilogram of diet for 6 weeks. The hormone treatment produced significant decreases in the contraction time, one-half relaxation time, and twitch tension in the SOL, while the peak rate of tension development (+ dP/dt) and decline (- dP/dt) in this muscle were elevated. Additionally, the force-frequency curve was shifted to the right and, thus, resembled the curve of a normal fast-twitch muscle. In contrast, the contractile properties of the fast EDL were relatively unaltered by the hormone administration. Thyrotoxicosis also changed the SOL response to contractile activity as twitch tension, + dP/dt, and - dP/dt remained high, and a faster decline in muscle glycogen and an increase in lactate occurred compared to control muscles. These results clearly demonstrate a preferential effect of thyroid hormone on slow compared to fast skeletal muscle.


Adjuvant chemotherapy for gastric cancer with S-1, an oral fluoropyrimidine.
Advanced gastric cancer can respond to S-1, an oral fluoropyrimidine. We tested S-1 as adjuvant chemotherapy in patients with curatively resected gastric cancer. Patients in Japan with stage II or III gastric cancer who underwent gastrectomy with extended (D2) lymph-node dissection were randomly assigned to undergo surgery followed by adjuvant therapy with S-1 or to undergo surgery only. In the S-1 group, administration of S-1 was started within 6 weeks after surgery and continued for 1 year. The treatment regimen consisted of 6-week cycles in which, in principle, 80 mg of oral S-1 per square meter of body-surface area per day was given for 4 weeks and no chemotherapy was given for the following 2 weeks. The primary end point was overall survival. We randomly assigned 529 patients to the S-1 group and 530 patients to the surgery-only group between October 2001 and December 2004. The trial was stopped on the recommendation of the independent data and safety monitoring committee, because the first interim analysis, performed 1 year after enrollment was completed, showed that the S-1 group had a higher rate of overall survival than the surgery-only group (P=0.002). Analysis of follow-up data showed that the 3-year overall survival rate was 80.1% in the S-1 group and 70.1% in the surgery-only group. The hazard ratio for death in the S-1 group, as compared with the surgery-only group, was 0.68 (95% confidence interval, 0.52 to 0.87; P=0.003). Adverse events of grade 3 or grade 4 (defined according to the Common Toxicity Criteria of the National Cancer Institute) that were relatively common in the S-1 group were anorexia (6.0%), nausea (3.7%), and diarrhea (3.1%). S-1 is an effective adjuvant treatment for East Asian patients who have undergone a D2 dissection for locally advanced gastric cancer. (ClinicalTrials.gov number, NCT00152217 [ClinicalTrials.gov].).


Deletion of Kv4.2 gene eliminates dendritic A-type K+ current and enhances induction of long-term potentiation in hippocampal CA1 pyramidal neurons.
Dendritic, backpropagating action potentials (bAPs) facilitate the induction of Hebbian long-term potentiation (LTP). Although bAPs in distal dendrites of hippocampal CA1 pyramidal neurons are attenuated when propagating from the soma, their amplitude can be increased greatly via downregulation of dendritic A-type K+ currents. The channels that underlie these currents thus may represent a key regulatory component of the signaling pathways that lead to synaptic plasticity. We directly tested this hypothesis by using Kv4.2 knock-out mice. Deletion of the Kv4.2 gene and a loss of Kv4.2 protein resulted in a specific and near-complete elimination of A-type K+ currents from the apical dendrites of CA1 pyramidal neurons. The absence of dendritic Kv4.2-encoded A-type K+ currents led to an increase of bAP amplitude and an increase of concurrent Ca2+ influx. Furthermore, CA1 pyramidal neurons lacking dendritic A-type K+ currents from Kv4.2 knock-out mice exhibited a lower threshold than those of wild-type littermates for LTP induction with the use of a theta burst pairing protocol. LTP triggered with the use of a saturating protocol, on the other hand, remained indistinguishable between Kv4.2 knock-out and wild-type neurons. Our results support the hypothesis that dendritic A-type K+ channels, composed of Kv4.2 subunits, regulate action potential backpropagation and the induction of specific forms of synaptic plasticity.


Fiber-based stray light suppression in spectroscopy using periodic shadowing.
Stray light is a known strong interference in spectroscopic measurements. Photons from high-intensity signals that are scattered inside the spectrometer, or photons that enter the detector through unintended ways, will be added to the spectrum as an interference signal. A general experimental solution to this problem is presented here by introducing a customized fiber for signal collection. The fiber-mount to the spectrometer consists of a periodically arranged fiber array that, combined with lock-in analysis of the data, is capable of suppressing stray light for improved spectroscopy. The method, which is referred to as fiber-based periodic shadowing, was applied to Raman spectroscopy in combustion. The fiber-based stray-light suppression method is implemented in an experimental setup with a high-power high-repetition-rate laser system used for Raman measurements in different room-temperature gas mixtures and a premixed flame. It is shown that the stray-light level is reduced by up to a factor of 80. Weak spectral lines can be distinguished, and therefore better molecular species identification, as well as concentration and temperature evaluation, were performed. The results show that the method is feasible and efficient in practical use and that it can be employed as a general tool for improving spectroscopic accuracy.


Safety and efficacy of galantamine (Reminyl) in severe Alzheimer's disease (the SERAD study): a randomised, placebo-controlled, double-blind trial.
The efficacy of galantamine has been shown in patients with mild, moderate, and advanced moderate Alzheimer's disease (AD). Here we report its efficacy in patients with severe AD. Between December, 2003, and March, 2007, patients aged 84 (SD 6) years with severe AD (mini-mental state examination [MMSE] score 5-12 points), in a nursing home setting were randomly assigned to receive galantamine (n=207), titrated initially to 24 mg/day, or placebo (n=200). Co-primary efficacy measures for cognitive function and ability to undertake normal daily activities were the severe impairment battery (SIB) and the seven-item minimum data set-activities of daily living (MDS-ADL), respectively. Adverse events, vital signs, laboratory parameters, and electrocardiograms were monitored. This trial is registered with ClinicalTrials.gov, number NCT00216593. 168 of 207 (81%) patients in the galantamine group and 161 of 200 (81%) in the placebo group completed the study. Mean SIB scores increased (improved) by 1.9 (95% CI -0.1 to 3.9) points with galantamine and decreased (worsened) by 3.0 (-5.6 to -0.5) points with placebo (between-group least squares mean difference 4.36, 1.3 to 7.5; p=0.006). Mean MDS-ADL self-performance score worsened by 1.2 (0.6 to 1.8) points and 1.6 (0.8 to 2.3) points, respectively (between-group least squares mean difference -0.41, -1.3 to 0.5; p=0.383). Nominally significant between-group differences in favour of galantamine occurred for the SIB domains of memory (p=0.006), praxis (p=0.010), and visuospatial ability (p=0.002), and for the MDS-ADL subitem locomotion on unit (p=0.021). 183 of 207 patients (88%) who received galantamine and 177 of 200 (89%) who received placebo had adverse events, which were mostly mild to moderate. Eight patients (4%) in the galantamine group and 21 patients (11%) in the placebo group died. ECG abnormalities were similar between the two groups. Galantamine can be started and used safely in elderly patients with severe AD. Galantamine improved cognitive function but failed to significantly improve the co-primary parameter of overall activities of daily living.


Calibration maintenance and transfer using Tikhonov regularization approaches.
Maintaining multivariate calibrations is essential and involves keeping models developed on an instrument applicable to predicting new samples over time. Sometimes a primary instrument model is needed to predict samples measured on secondary instruments. This situation is referred to as calibration transfer. This paper reports on using a Tikhonov regularization (TR) based method in both cases. A distinction of the TR design for calibration maintenance and transfer is a defined weighting scheme for a small set of new (transfer or standardization) samples augmented to the full set of calibration samples. Because straight application of basic TR theory is not always possible with calibration maintenance and transfer, this paper develops a generic solution to always enable application of TR. Harmonious (bias/variance tradeoff) and parsimonious (effective rank) considerations for TR are compared with the same TR format applied to partial least squares (PLS), showing that both approaches are viable solutions to the calibration maintenance and transfer problems.


Investigation of the structural requirements for N-methyl-D-aspartate receptor positive and negative allosteric modulators based on 2-naphthoic acid.
The N-methyl-D-aspartate receptor (NMDAR), a ligand-gated ion channel activated by L-glutamate and glycine, plays a major role in the synaptic plasticity underlying learning and memory. NMDARs are involved in neurodegenerative disorders such as Alzheimer's and Parkinson's disease and NMDAR hypofunction is implicated in schizophrenia. Herein we describe structure-activity relationship (SAR) studies on 2-naphthoic acid derivatives to investigate structural requirements for positive and negative allosteric modulation of NMDARs. These studies identified compounds such as UBP684 (14b), which act as pan potentiators by enhancing NMDAR currents in diheteromeric NMDAR tetramers containing GluN1 and GluN2A-D subunits. 14b and derivatives thereof are useful tools to study synaptic function and have potential as leads for the development of drugs to treat schizophrenia and disorders that lead to a loss of cognitive function. In addition, SAR studies have identified a series of styryl substituted compounds with partial NAM activity and a preference for inhibition of GluN2D versus the other GluN2 subunits. In particular, the 3-and 2-nitrostyryl derivatives UBP783 (79i) and UBP792 (79h) had IC50s of 1.4 μM and 2.9 μM, respectively, for inhibition of GluN2D but showed only 70-80% maximal inhibition. GluN2D has been shown to play a role in excessive pain transmission due to nerve injury and potentially in neurodegenerative disorders. Partial GluN2D inhibitors may be leads for the development of drugs to treat these disorders without the adverse effects observed with full NMDAR antagonists.


Serum laminin, hydrocarbon exposure, and glomerular damage.
It has been postulated that occupational exposure to hydrocarbons may damage the kidney and lead to glomerulonephritis and chronic renal failure. As laminin is a ubiquitous basement membrane component that seems to play a central part in the structure and function of basement membranes and as the normal renal filtration process is highly dependent on an intact glomerular basement membrane, the serum laminin concentration was examined in a population of workers exposed to hydrocarbons. The hydrocarbon exposure was assessed by exposure surrogates (exposure duration and exposure score). An interaction between occupational exposure to hydrocarbons and hypertension increased the laminin concentration whereas the laminin concentration decreased in workers exposed for a long time probably because of a selection effect. In a subgroup of printers exposed to toluene whose hippuric acid excretion had been recorded for several years this interaction was confirmed when the hippuric acid excretion was substituted for the other exposure indices. In the exposed group, the age-related decline in creatinine clearance was accelerated. These results seem to confirm that occupational exposure to hydrocarbons is a non-specific factor that may promote a deterioration of renal function.


Fiber-Optic Intracranial Pressure Monitoring System Using Wi-Fi-An In Vivo Study.
Continuous invasive monitoring of intracranial pressure (ICP) is essential in neurocritical care for surveillance and management of raised ICP. Fluid-based systems and strain gauge microsensors remain the current standard. In the past few decades, several studies with wireless monitoring were developed aiming to reduce invasiveness and complications. To describe a novel Wi-Fi fiber-optic device for continuous ICP monitoring using smartphone in a swine model. Two ICP sensors (wireless prototype and wire-based reference) were implanted in the cerebral parenchyma of a swine model for a total of 120 minutes of continuous monitoring. Every 5 minutes, jugular veins compression was performed to evaluate ICP changes. The experimentation was divided in 3 phases for comparison and analysis. Phase 1 showed agreement in ICP changes for both sensors during jugular compression and releasing, with a positive and strong Spearman correlation (r = 0.829, P < .001). Phase 2 started after inversion of the sensors in the burr holes; there was a positive and moderately weak Spearman correlation (r = 0.262, P < .001). For phase 3, the sensors were returned to the first burr holes; the prototype behaved similarly to the reference sensor, presenting a positive and moderately strong Spearman correlation (r = 0.669, P < .001). A Wi-Fi ICP monitoring system was demonstrated in a comprehensive and feasible way. It was possible to observe, using smartphone, an adequate correlation regarding ICP variations. Further adaptations are already being developed.


Mild renal dysfunction and metabolites tied to low HDL cholesterol are associated with monocytosis and atherosclerosis.
The number of circulating blood monocytes impacts atherosclerotic lesion size, and in mouse models, elevated levels of high-density lipoprotein cholesterol suppress blood monocyte counts and atherosclerosis. We hypothesized that individuals with mild renal dysfunction at increased cardiovascular risk would have reduced high-density lipoprotein levels, high blood monocyte counts, and accelerated atherosclerosis. To test whether mild renal dysfunction is associated with an increase in a leukocyte subpopulation rich in monocytes that has a known association with future coronary events, we divided individuals from the Malmö Diet and Cancer study (MDC) into baseline cystatin C quintiles (n=4757). Lower levels of renal function were accompanied by higher monocyte counts, and monocytes were independently associated with carotid bulb intima-media thickness cross-sectionally (P=0.02). Cystatin C levels were positively and plasma high-density lipoprotein cholesterol levels negatively associated with monocyte counts at baseline, after adjustment for traditional risk factors. Several amino acid metabolites tied to low levels of high-density lipoprotein cholesterol and insulin resistance measured in a subset of individuals (n=752) by use of liquid chromatography-mass spectrometry were independently associated with a 22% to 34% increased risk of being in the top quartile of monocytes (P<0.05). A low high-density lipoprotein cholesterol, insulin resistance phenotype occurs in subjects with mild renal dysfunction and is associated with elevated monocytes and atherosclerosis. High blood monocyte counts may represent a previously unrecognized mechanism underlying the strong relationship between cystatin C and cardiovascular risk.


The interplay between single particle anisotropy and interparticle interactions in ensembles of magnetic nanoparticles.
This paper aims to analyze the competition of single particle anisotropy and interparticle interactions in nanoparticle ensembles using a random anisotropy model. The model is first applied to ideal systems of non-interacting and strongly dipolar interacting ensembles of maghemite nanoparticles. The investigation is then extended to more complex systems of pure cobalt ferrite CoFe2O4 (CFO) and mixed cobalt-nickel ferrite (Co,Ni)Fe2O4 (CNFO) nanoparticles. Both samples were synthetized by the polyol process and exhibit the same particle size (DTEM ≈ 5 nm), but with different interparticle interaction strengths and single particle anisotropy. The implementation of the random anisotropy model allows investigation of the influence of single particle anisotropy and interparticle interactions, and sheds light on their complex interplay as well as on their individual contribution. This analysis is of fundamental importance in order to understand the physics of these systems and to develop technological applications based on concentrated magnetic nanoparticles, where single and collective behaviors coexist.


Architectural Changes of the Biceps Femoris Long Head after Concentric or Eccentric Training.
To determine the architectural adaptations of the biceps femoris long head (BFlh) after concentric or eccentric strength training interventions and the time course of adaptation during training and detraining. Participants in this intervention (concentric training group [n = 14], eccentric training group [n = 14], male subjects) completed a 4-wk control period, followed by 6 wk of either concentric- or eccentric-only knee flexor training on an isokinetic dynamometer and finished with 28 d of detraining. Architectural characteristics of BFlh were assessed at rest and during graded isometric contractions using two-dimensional ultrasonography at 28 d prebaseline; baseline; and days 14, 21, and 42 of the intervention and then again after 28 d of detraining. BFlh fascicle length was significantly longer in the eccentric training group (P < 0.05; d range, 2.65-2.98) and shorter in the concentric training group (P < 0.05; d range, -1.62 to -0.96) after 42 d of training compared with baseline at all isometric contraction intensities. After the 28-d detraining period, BFlh fascicle length was significantly reduced in the eccentric training group at all contraction intensities compared with the end of the intervention (P < 0.05; d range, -1.73 to -1.55). There was no significant change in fascicle length of the concentric training group after the detraining period. These results provide evidence that short-term resistance training can lead to architectural alterations in the BFlh. In addition, the eccentric training-induced lengthening of BFlh fascicle length was reversed and returned to baseline values after 28 d of detraining. The contraction mode specific adaptations in this study may have implications for injury prevention and rehabilitation.


Self-assembly of vascularized tissue to support tumor explants in vitro.
Testing the efficacy of cancer drugs requires functional assays that recapitulate the cell populations, anatomy and biological responses of human tumors. Although current animal models and in vitro cell culture platforms are informative, they have significant shortcomings. Mouse models can reproduce tissue-level and systemic responses to tumor growth and treatments observed in humans, but xenografts from patients often do not grow, or require months to develop. On the other hand, current in vitro assays are useful for studying the molecular bases of tumorigenesis or drug activity, but often lack the appropriate in vivo cell heterogeneity and natural microenvironment. Therefore, there is a need for novel tools that allow rapid analysis of patient-derived tumors in a robust and representative microenvironment. We have developed methodology for maintaining harvested tumor tissue in vitro by placing them in a support bed with self-assembled stroma and vasculature. The harvested biopsy or tumor explant integrates with the stromal bed and vasculature, providing the correct extracellular matrix (collagen I, IV, fibronectin), associated stromal cells, and a lumenized vessel network. Our system provides a new tool that will allow ex vivo drug-screening and can be adapted for the guidance of patient-specific therapeutic strategies.


Non-pharmacological interventions for wandering of people with dementia in the domestic setting.
A number of studies exist of interventions for wandering in the institutional setting, but much less work has been done on wandering in the domestic setting. The prevalence of wandering by people with dementia is difficult to assess; wandering is not a simple or static behaviour and the reasons why people wander remain unclear. In the absence of a theory of wandering and an agreed definition of wandering, it is difficult to discover effective strategies for managing wandering and difficult to design appropriate intervention strategies. Also, the same behaviour or type of wandering might occur for different reasons in different individuals; any theoretical formulation is going to have to allow for different triggers for the behaviour and so to get a 'one size fits all' kind of explanation is unlikely. Thus what we mostly encounter in this field is a 'trial and error' approach which does not always do justice to the complex interactions of personal and environmental factors that lead people with dementia to wander. While there seems to be a consensus in the literature that in the majority of cases non-pharmacological approaches may work as well as drug treatment and with fewer side effects, in practice clinicians often resort to drugs as the first line of treatment. This review reports the lack of evidence from RCTs and discusses the range of non-pharmacological interventions that have been carried out using other study designs. To evaluate the effectiveness and safety of non-pharmacological interventions in reducing wandering in the domestic setting by people with dementia. The secondary objective is to highlight the quality and quantity of research evidence available and to set an agenda for future research. The trials were identified from a search of the Specialized Register of the Cochrane Dementia and Cognitive Improvement Group on 11 May 2006 using the terms exit* or wander* or elopement or ambulat* or walk*. This Register contains records from all major healthcare databases and many ongoing trial databases and is updated regularly. Randomised clinical trials comparing intervention with no intervention or usual treatment ('standard care') or another intervention. No suitable trials of non-pharmacological interventions for the prevention and management of wandering in the domestic setting were found. As no randomised controlled trials were found, no results can be reported. There is an urgent need for randomised controlled trials of non-pharmacological interventions for wandering in the domestic setting.


[BCcogSEP: a French test battery evaluating cognitive functions in multiple sclerosis].
In French language, there is no standardized procedure to assess cognitive function in patients with multiple sclerosis (MS). Such an assessment is however very useful to determine the consequences of the disease on cognitive function, to evaluate the disease progression and the consequences of usual treatments on cognition. This study aimed to develop and validate a French language battery based on "the Brief Repeatable Battery of Neuropsychological tests for Multiple Sclerosis" (BRB-N) often used in other countries. In an initial phase, the battery was composed of the French version of the 5 BRB-N tests to which were added 4 tests assessing immediate and working memory as well as executive function. 52 healthy control subjects (20-50 years) participated in the study. A principal component analysis (PCA) of their data examined the contribution of each test into the battery. 93 other healthy subjects participated in a second phase where analyses of variance were carried out to investigate the effect of the main demographical variables. Finally, the performance of two patient groups (20 with a EDSS score<or=3 et 31 with a EDSS score>3) was compared to the healthy controls in order to investigate the ability of our battery to detect cognitive impairment in MS patients. The PCA showed that each test, except one, had a specific contribution to the battery. The final battery (BCcogSEP) was thus comprised of 8 tests. MANOVA and ANOVA showed significant effects of age, sex and educational level on performance. In consequence, a procedure allowing to take into account these factors was developed. The battery was able to detect cognitive impairment in MS patients, even when the disease is not severe. In this case, deficits were observed in tests assessing executive function, information processing speed, immediate and working memory. This short battery with reliable psychometric qualities allows the interpretation of a MS patient's performance considering his/her demographical characteristics and is able to detect cognitive abnormalities even in case of mild physical handicap.


Multimedia computer-assisted instruction in cardiology.
Medical education faces problems caused by increasing restraints on resources. A multicenter consortium combined simulation and multimedia computer-assisted instruction (MCAI) to develop unique interactive teaching programs that can address a number of these problems. We describe the consortium, the MCAI system, the programs, and a multicenter evaluation of technical and educational performance. The MCAI system uses computer-controlled access to full-screen, full-motion, and full-color laser disc video in combination with digitized sound, images, and graphics stored on removable media. The Socratic teaching method enhances interaction and guides learners through the patient's history, cardiovascular physical examination, laboratory evaluation, and therapy. Self-instruction and instructor-led modes of function are possible. The first five programs, based on simulations of specific cardiology problems, were distributed to four medical centers. Questionnaires evaluated technical function and medical student opinions, while behaviors and scores were automatically tracked and tabulated by program administration software. The MCAI system functioned reliably and accurately in all modes and at all sites. The programs were highly rated. Student ratings, scores, and behaviors were independent of institution and mode of use. A multicenter educational consortium developed a system to produce unique, sophisticated MCAI programs in cardiology. Both system and programs functioned reliably at four institutions and were highly rated by fourth-year medical students. With this enthusiastic reception, the economies and strengths associated with MCAI make it an attractive solution to a number of problem areas, and it will likely play an increasingly important role.


An automated shotgun lipidomics platform for high throughput, comprehensive, and quantitative analysis of blood plasma intact lipids.
Blood plasma has gained protagonism in lipidomics studies due to its availability, uncomplicated collection and preparation, and informative readout of physiological status. At the same time, it is also technically challenging to analyze due to its complex lipid composition affected by many factors, which can hamper the throughput and/or lipidomics coverage. To tackle these issues, we developed a comprehensive, high throughput, and quantitative mass spectrometry-based shotgun lipidomics platform for blood plasma lipid analyses. The main hallmarks of this technology are (i) it is comprehensive, covering 22 quantifiable different lipid classes encompassing more than 200 lipid species; (ii) it is amenable to high-throughput, with less than 5 min acquisition time allowing the complete analysis of 200 plasma samples per day; (iii) it achieves absolute quantification, by inclusion of internal standards for every lipid class measured; (iv) it is highly reproducible, achieving an average coefficient of variation of <10% (intra-day), approx. 10% (inter-day), and approx. 15% (inter-site) for most lipid species; (v) it is easily transferable allowing the direct comparison of data acquired in different sites. Moreover, we thoroughly assessed the influence of blood stabilization with different anticoagulants and freeze-thaw cycles to exclude artifacts generated by sample preparation. Practical applications: This shotgun lipidomics platform can be implemented in different laboratories without compromising reproducibility, allowing multi-site studies and inter-laboratory comparisons. This possibility combined with the high-throughput, broad lipidomic coverage and absolute quantification are important aspects for clinical applications and biomarker research.


Comparison of CpG island methylator phenotype (CIMP) frequency in colon cancer using different probe- and gene-specific scoring alternatives on recommended multi-gene panels.
In colorectal cancer a distinct subgroup of tumours demonstrate the CpG island methylator phenotype (CIMP). However, a consensus of how to score CIMP is not reached, and variation in definition may influence the reported CIMP prevalence in tumours. Thus, we sought to compare currently suggested definitions and cut-offs for methylation markers and how they influence CIMP classification in colon cancer. Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA), with subsequent fragment analysis, was used to investigate methylation of tumour samples. In total, 31 CpG sites, located in 8 different genes (RUNX3, MLH1, NEUROG1, CDKN2A, IGF2, CRABP1, SOCS1 and CACNA1G) were investigated in 64 distinct colon cancers and 2 colon cancer cell lines. The Ogino gene panel includes all 8 genes, in addition to the Weisenberger panel of which only 5 of the 8 genes included were investigated. In total, 18 alternative combinations of scoring of CIMP positivity on probe-, gene-, and panel-level were analysed and compared. For 47 samples (71%), the CIMP status was constant and independent of criteria used for scoring; 34 samples were constantly scored as CIMP negative, and 13 (20%) consistently scored as CIMP positive. Only four of 31 probes (13%) investigated showed no difference in the numbers of positive samples using the different cut-offs. Within the panels a trend was observed that increasing the gene-level stringency resulted in a larger difference in CIMP positive samples than increasing the probe-level stringency. A significant difference between positive samples using 'the most stringent' as compared to 'the least stringent' criteria (20% vs 46%, respectively; p<0.005) was demonstrated. A statistical significant variation in the frequency of CIMP depending on the cut-offs and genes included in a panel was found, with twice as many positives samples by least compared to most stringent definition used.


Persistence of goitre in children post-salt iodization in Islamic Republic of Iran: autoimmune status.
We evaluated the prevalence of autoimmune thyroiditis in a random sample of 1188 schoolchildren aged 8-13 years with normalized iodine intake in the Islamic Republic of Iran. The prevalence of goitre was 39.6%; the majority had palpable but non-visible goitre. Of a subsample of 500 children, median urinary iodine excretion (18/8 microg/dL) indicated normal iodine intake. Thyroid peroxidase (TPO) antibody was positive in 3.7% of children and was significantly correlated with the prevalence of goitre and hypothyroidism. No correlation was seen between urinary iodine excretion and positive TPO antibody, mean TPO antibody, hypothyroidism or prevalence of goitre. Autoimmune thyroiditis explains some cases of goitre but other goitrogenic factors need to be evaluated.


Vaccine Containing the Three Allelic Variants of the Plasmodium vivax Circumsporozoite Antigen Induces Protection in Mice after Challenge with a Transgenic Rodent Malaria Parasite.
Plasmodium vivax is the most common species that cause malaria outside of the African continent. The development of an efficacious vaccine would contribute greatly to control malaria. Recently, using bacterial and adenoviral recombinant proteins based on the P. vivax circumsporozoite protein (CSP), we demonstrated the possibility of eliciting strong antibody-mediated immune responses to each of the three allelic forms of P. vivax CSP (PvCSP). In the present study, recombinant proteins representing the PvCSP alleles (VK210, VK247, and P. vivax-like), as well as a hybrid polypeptide, named PvCSP-All epitopes, were generated. This hybrid containing the conserved C-terminal of the PvCSP and the three variant repeat domains in tandem were successfully produced in the yeast Pichia pastoris. After purification and biochemical characterization, they were used for the experimental immunization of C57BL/6 mice in a vaccine formulation containing the adjuvant Poly(I:C). Immunization with a recombinant protein expressing all three different allelic forms in fusion elicited high IgG antibody titers reacting with all three different allelic variants of PvCSP. The antibodies targeted both the C-terminal and repeat domains of PvCSP and recognized the native protein on the surface of P. vivax sporozoites. More importantly, mice that received the vaccine formulation were protected after challenge with chimeric Plasmodium berghei sporozoites expressing CSP repeats of P. vivax sporozoites (Pb/PvVK210). Our results suggest that it is possible to elicit protective immunity against one of the most common PvCSP alleles using soluble recombinant proteins expressed by P. pastoris. These recombinant proteins are promising candidates for clinical trials aiming to develop a multiallele vaccine against P. vivax malaria.


Hypothalamic JNK1-hepatic fatty acid synthase axis mediates a metabolic rewiring that prevents hepatic steatosis in male mice treated with olanzapine via intraperitoneal: Additional effects of PTP1B inhibition.
Olanzapine (OLA), a widely used second-generation antipsychotic (SGA), causes weight gain and metabolic alterations when administered orally to patients. Recently, we demonstrated that, contrarily to the oral treatment which induces weight gain, OLA administered via intraperitoneal (i.p.) in male mice resulted in body weight loss. This protection was due to an increase in energy expenditure (EE) through a mechanism involving the modulation of hypothalamic AMPK activation by higher OLA levels reaching this brain region compared to those of the oral treatment. Since clinical studies have shown hepatic steatosis upon chronic treatment with OLA, herein we further investigated the role of the hypothalamus-liver interactome upon OLA administration in wild-type (WT) and protein tyrosine phosphatase 1B knockout (PTP1B-KO) mice, a preclinical model protected against metabolic syndrome. WT and PTP1B-KO male mice were fed an OLA-supplemented diet or treated via i.p. Mechanistically, we found that OLA i.p. treatment induces mild oxidative stress and inflammation in the hypothalamus in a JNK1-independent and dependent manner, respectively, without features of cell dead. Hypothalamic JNK activation up-regulated lipogenic gene expression in the liver though the vagus nerve. This effect concurred with an unexpected metabolic rewiring in the liver in which ATP depletion resulted in increased AMPK/ACC phosphorylation. This starvation-like signature prevented steatosis. By contrast, intrahepatic lipid accumulation was observed in WT mice treated orally with OLA; this effect being absent in PTP1B-KO mice. We also demonstrated an additional benefit of PTP1B inhibition against hypothalamic JNK activation, oxidative stress and inflammation induced by chronic OLA i.p. treatment, thereby preventing hepatic lipogenesis. The protection conferred by PTP1B deficiency against hepatic steatosis in the oral OLA treatment or against oxidative stress and neuroinflammation in the i.p. treatment strongly suggests that targeting PTP1B might be also a therapeutic strategy to prevent metabolic comorbidities in patients under OLA treatment in a personalized manner.


Application of a new leaf area index algorithm to China's landmass using MODIS data for carbon cycle research.
An operational system was developed for mapping the leaf area index (LAI) for carbon cycle models from the moderate resolution imaging spectroradiometer (MODIS) data. The LAI retrieval algorithm is based on Deng et al. [2006. Algorithm for global leaf area index retrieval using satellite imagery. IEEE Transactions on Geoscience and Remote Sensing, 44, 2219-2229], which uses the 4-scale radiative transfer model [Chen, J.M., Leblancs, 1997. A 4-scale bidirectional reflection model based on canopy architecture. IEEE Transactions on Geoscience and Remote Sensing, 35, 1316-1337] to simulate the relationship of LAI with vegetated surface reflectance measured from space for various spectral bands and solar and view angles. This algorithm has been integrated to the MODISoft platform, a software system designed for processing MODIS data, to generate 250 m, 500 m and 1 km resolution LAI products covering all of China from MODIS MOD02 or MOD09 products. The multi-temporal interpolation method was implemented to remove the residual cloud and other noise in the final LAI product so that it can be directly used in carbon models without further processing. The retrieval uncertainties from land cover data were evaluated using five different data sets available in China. The results showed that mean LAI discrepancies can reach 27%. The current product was also compared with the NASA MODIS MOD15 LAI product to determine the agreement and disagreement of two different product series. LAI values in the MODIS product were found to be 21% larger than those in the new product. These LAI products were compared against ground TRAC measurements in forests in Qilian Mountain and Changbaishan. On average, the new LAI product agrees with the field measurement in Changbaishan within 2%, but the MODIS product is positively biased by about 20%. In Qilian Mountain, where forests are sparse, the new product is lower than field measurements by about 38%, while the MODIS product is larger by about 65%.


Existence of different Fos/Jun complexes during the G0-to-G1 transition and during exponential growth in mouse fibroblasts: differential role of Fos proteins.
We have determined the different Fos/Jun complexes present in Swiss 3T3 cells either following serum stimulation of quiescent cells or during exponential growth by immunoprecipitation analyses. We have shown that while c-Fos is the major Fos protein associated with the Jun proteins (c-Jun, JunB, and JunD) soon after serum stimulation, at later times Fra-1 and Fra-2 are the predominant Fos proteins associated with the different Jun proteins. During exponential growth, the synthesis of Fra-1 and Fra-2 is maintained at a significant level, in contrast to c-Fos and FosB, which are expressed at very low or undetectable levels. Consequently, Fra-1 and Fra-2 are the main Fos proteins complexed with the Jun proteins in asynchronously growing cells. To determine whether the Fos proteins are differentially required during the G0-to-G1 transition and exponential growth for the entrance into S phase, we microinjected affinity-purified antibodies directed against c-Fos, FosB, Fra-1, and Fra-2. We have found that while the activities of c-Fos and FosB are required mostly during the G0-to-G1 transition, Fra-1 and Fra-2 are involved both in the G0-to-G1 transition and in asynchronous growth.


Chemical agents transported by xylem mass flow propagate variation potentials.
Long-distance signalling is important for coordinating plant responses to the environment. Variation potentials (VPs) are a type of long-distance electrical signal that are generated in plants in response to wounding or flaming. Unlike self-propagating action potentials, VPs can be measured beyond regions of dead or chemically treated tissue that block signal generation, suggesting a different mode of propagation. Two alternative propagation mechanisms have been proposed: movement of a chemical agent and a pressure wave through the vasculature. Variants of these two signalling mechanisms have been suggested. Here, we use simple models of the underlying physical processes to evaluate and compare these predictions against independent data. Our models suggest that chemical diffusion and pressure waves are unlikely to capture existing data with parameters that are known from other sources. The previously discarded hypothesis of mass flow in the xylem transporting a chemical agent, however, is able to reproduce experimental propagation speeds for VPs. We therefore suggest that chemical agents transported by mass flow within the xylem are more likely than a pressure wave or chemical diffusion as a VP propagation mechanism. Understanding this mode of long-distance signalling within plants is important for unravelling how plants coordinate physiological responses via cell-to-cell communication.


Epigenetic regulation of the oxytocin receptor gene: implications for behavioral neuroscience.
Genetic approaches have improved our understanding of the neurobiological basis of social behavior and cognition. For instance, common polymorphisms of genes involved in oxytocin signaling have been associated with sociobehavioral phenotypes in healthy samples as well as in subjects with mental disorders. More recently, attention has been drawn to epigenetic mechanisms, which regulate genetic function and expression without changes to the underlying DNA sequence. We provide an overview of the functional importance of oxytocin receptor gene (OXTR) promoter methylation and summarize studies that have investigated the role of OXTR methylation in behavioral phenotypes. There is first evidence that OXTR methylation is associated with autism, high callous-unemotional traits, and differential activation of brain regions involved in social perception. Furthermore, psychosocial stress exposure might dynamically regulate OXTR. Given evidence that epigenetic states of genes can be modified by experiences, especially those occurring in sensitive periods early in development, we conclude with a discussion on the effects of traumatic experience on the developing oxytocin system. Epigenetic modification of genes involved in oxytocin signaling might be involved in the mechanisms mediating the long-term influence of early adverse experiences on socio-behavioral outcomes.


Product of the Pseudomonas aeruginosa gene pilD is a prepilin leader peptidase.
The related type IV pilins produced by Pseudomonas aeruginosa, Neisseria gonorrhoeae, Bacteroides nodosus, and Moraxella bovis are synthesized as precursors with short, six- or seven-amino acid N-terminal leader peptides. We have previously observed that P. aeruginosa mutations in pilD, a gene required for pilus biogenesis, result in the accumulation of unprocessed prepilin in the membrane and a general defect in the excretion of a number of extracellular enzymes. An endopeptidase activity has been detected in detergent-solubilized inner membrane of P. aeruginosa and shown to correctly cleave the prepilin of P. aeruginosa and N. gonorrhoeae. It is absent from pilD mutants, increased by pilD overexpression, and conferred on Escherichia coli by the introduction of the pilD gene. The pilD gene product, purified by immunoaffinity chromatography with antibody to a PilD-derived synthetic peptide, was identified with the endopeptidase. PilD appears to be a prototype of a class of enzymes that process not only type IV pilin precursors but also components of a protein-excretion apparatus of Gram-negative bacteria.


IgE antibodies to alpha-gal in the general adult population: relationship with tick bites, atopy, and cat ownership.
The carbohydrate alpha-gal epitope is present in many animal proteins, including those of red meat and animal immunoglobulins, such as cat IgA. Systemic anaphylaxis to the alpha-gal epitope has recently been described. To investigate and compare the prevalence of alpha-gal-specific (s)IgE and its associated factors in the general adult population from two separated (Northern and Southern) European regions (Denmark and Spain, respectively). Cross-sectional study of 2297 and 444 randomly selected adults from 11 municipalities in Denmark and one in Spain. Alpha-gal sIgE was assessed by ImmunoCAP to bovine thyroglobulin. Additional assessments included a panel of skin prick test (SPT) to common aeroallergens and epidemiological factors, including the history of tick bites in the Danish series. The prevalence of positive (≥ 0.1 kUA /L) sIgE to alpha-gal was 5.5% and 8.1% in the Danish and Spanish series, respectively. The prevalence of sIgE ≥ 0.35 kUA /L was 1.8% and 2.2% in Denmark and Spain, respectively. Alpha-gal sIgE positivity was associated with pet ownership in both series and, particularly, cat ownership (data available in the Danish series). Alpha-gal sIgE positivity was associated with atopy (SPT positivity) in both series, although it was not associated with SPT positivity to cat or dog dander. Alpha-gal sIgE positivity was strongly associated with a history of tick bites. The prevalence of alpha-gal sIgE antibodies in these general adult European populations is similarly low. The presence of alpha-gal sIgE antibodies is associated with a history of tick bites, atopy, and cat ownership.


Heroin detoxification during pregnancy: A systematic review and retrospective study of the management of heroin addiction in pregnancy.
There is general consensus that methadone maintenance is the gold standard in the management of pregnant heroin users. However, in South African state hospitals, methadone withdrawal is the routine procedure offered to these patients, as methadone maintenance programmes are unavailable in the public sector. To conduct a systematic review of the literature on heroin detoxification in pregnancy, and to document pregnancy outcomes in heroin users detoxified with methadone at Groote Schuur Hospital (GSH), Cape Town, from 2006 to 2010. A literature search was undertaken to identify key publications on the management of heroin addiction in pregnancy. Patients for the study were identified from the GSH methadone registry, and data were collected from the clinical files. A total of 20 relevant publications were identified and reviewed. Early case reports described an increased risk of stillbirths and fetal distress after methadone detoxification, but more recent case series involving larger numbers of patients showed positive outcomes. In our study, six pregnant patients received methadone withdrawal over a 5-year period at GSH, and all the neonates had good Apgar scores and were discharged home within 3 days of delivery. There is limited evidence on the management of heroin addiction during pregnancy, and the only two guidelines identified suggest that methadone maintenance is preferable to methadone withdrawal. The favourable pregnancy outcomes in this small sample of patients managed with methadone withdrawal suggest that it may be safe and deserves further study.


Does body posture influence hand preference in an ancestral primate model?
The origin of human handedness and its evolution in primates is presently under debate. Current hypotheses suggest that body posture (postural origin hypothesis and bipedalism hypothesis) have an important impact on the evolution of handedness in primates. To gain insight into the origin of manual lateralization in primates, we studied gray mouse lemurs, suggested to represent the most ancestral primate condition. First, we investigated hand preference in a simple food grasping task to explore the importance of hand usage in a natural foraging situation. Second, we explored the influence of body posture by applying a forced food grasping task with varying postural demands (sit, biped, cling, triped). The tested mouse lemur population did not prefer to use their hands alone to grasp for food items. Instead, they preferred to pick them up using a mouth-hand combination or the mouth alone. If mouth usage was inhibited, they showed an individual but no population level handedness for all four postural forced food grasping tasks. Additionally, we found no influence of body posture on hand preference in gray mouse lemurs. Our results do not support the current theories of primate handedness. Rather, they propose that ecological adaptation indicated by postural habit and body size of a given species has an important impact on hand preference in primates. Our findings suggest that small-bodied, quadrupedal primates, adapted to the fine branch niche of dense forests, prefer mouth retrieval of food and are less manually lateralized than large-bodied species which consume food in a more upright, and less stable body posture.


[Effect of puerarin in myocardial protection in rats with acute and chronic alcoholism].
To investigate the protective effect of puerarin on the myocardium of rats with acute and chronic alcoholism. In acute alcoholism experiment, normal male SD rats were randomly divided into the control group, alcoholism group and puerarin group (n=8), and high- and low-dose puerarin was administered. In chronic alcoholism experiment, increasing puerarin doses were given. Serum and myocardial levels of spartate aminotransferase (AST) and creatine phosphokinase (CPK) were determined using enzymatic methed, and superoxide dismutase (SOD), malondialdehyde (MDA), Ca(2+)-Mg(2+)-ATPase, and Na(+)-K(+)-ATPase in the myocardium were assayed with colorimetric method. HE staining was used to observe the microscopic changes of the myocardium. Compared with alcoholism group, puerarin-treated groups showed significantly lowered myocardial contents of MDA, CPK and AST and serum levels of AST and CPK (P<0.05, P<0.01) and increased myocardial SOD (P<0.05, P<0.01), Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase activity (P<0.05, P<0.01), but Na(+)-K(+)-ATPase was similar between the two groups (P>0.05). HE staining of the myocardium showed cell swelling and obscure cell boundaries in alcoholism group, especially in chronic alcoholism group. The myocardial structure in puerarin group remained clear and regular. Puerarin can protect from myocardial injuries induced by acute and chronic alcoholism in rats.


Relapsing demyelinating disease affecting both the central and peripheral nervous systems.
Clinical and electromyographic findings of chronic inflammatory demyelinating polyradiculopathy (CIDP) are occasionally observed in patients with multiple sclerosis (MS). To define a new inflammatory demyelinating disease unlike MS or CIDP. This study reports on five patients with a demyelinating disease affecting the central nervous system (CNS) and peripheral nervous system (PNS). Each case presented a relapsing-remitting course in which CNS involvement preceded PNS involvement. All patients fulfilled Barkhof's criteria on MRI and the McDonald criteria for MS. Two patients had grey matter lesions with typical white matter changes. No systemic inflammatory disease and no metabolic or inflammatory factor for peripheral neuropathy were found. In all cases electromyography showed a demyelinating peripheral neuropathy without conduction block. Four patients fulfilled the European Federation of Neurological Societies/PNS guideline for CIDP and Nicolas et al's criteria for CIDP, one of whom also fulfilled the Ad Hoc Subcommittee criteria for CIDP. Nerve biopsy, performed in two patients, showed histological evidence of CIDP. An improvement in clinical status and neurophysiological parameters was observed in three patients after treatment with either intravenous immunoglobulin (n = 1) or cyclophosphamide (n = 2). The CNS and PNS demyelination, the absence of oligoclonal bands and the peripheral demyelination without conduction block indicate pathogenic mechanisms different from MS and CIDP. The chronology of events suggests an entity unlike that involved in acute demyelinating encephalomyelitis. Immunological reactivity against antigens common to peripheral and central myelin may explain why the demyelinating disease affected both the CNS and PNS.


Characterization of dark quencher chromophores as nonfluorescent acceptors for single-molecule FRET.
Dark quenchers are chromophores that primarily relax from the excited state to the ground state nonradiatively (i.e., are dark). As a result, they can serve as acceptors for Förster resonance energy transfer experiments without contributing significantly to background in the donor-emission channel, even at high concentrations. Although the advantages of dark quenchers have been exploited for ensemble bioassays, no systematic single-molecule study of dark quenchers has been performed, and little is known about their photophysical properties. Here, we present the first systematic single-molecule study of dark quenchers in conjunction with fluorophores and demonstrate the use of dark quenchers for monitoring multiple interactions and distances in multichromophore systems. Specifically, using double-stranded DNA standards labeled with two fluorophores and a dark quencher (either QSY7 or QSY21), we show that the proximity of a fluorophore and dark quencher can be monitored using the stoichiometry ratio available from alternating laser excitation spectroscopy experiments, either for single molecules diffusing in solution (using a confocal fluorescence) or immobilized on surfaces (using total-internal-reflection fluorescence). The latter experiments allowed characterization of the dark-quencher photophysical properties at the single-molecule level. We also use dark-quenchers to study the affinity and kinetics of binding of DNA Polymerase I (Klenow fragment) to DNA. The measured properties are in excellent agreement with the results of ensemble assays, validating the use of dark quenchers. Because dark-quencher-labeled biomolecules can be used in total-internal-reflection fluorescence experiments at concentrations of 1 μM or more without introducing a significant background, the use of dark quenchers should permit single-molecule Förster resonance energy transfer measurements for the large number of biomolecules that participate in interactions of moderate-to-low affinity.


The genetic basis of high-carbohydrate and high-monosodium glutamate diet related to the increase of likelihood of type 2 diabetes mellitus: a review.
Diabetes is one of the most common metabolic diseases. Aside from the genetic factor, previous studies stated that other factors such as environment, lifestyle, and paternal-maternal condition play critical roles in diabetes through DNA methylation in specific areas of the genome. One of diabetic cases is caused by insulin resistance and changing the homeostasis of blood glucose control so glucose concentration stood beyond normal rate (hyperglycemia). High fat diet has been frequently studied and linked to triggering diabetes. However, most Asians consume rice (or food with high carbohydrate) and food with monosodium glutamate (MSG). This habit could lead to pathophysiology of type 2 diabetes mellitus (T2D). Previous studies showed that high-carbohydrate or high-MSG diet could change gene expression or modify protein activity in body metabolism. This imbalanced metabolism can lead to pleiotropic effects of diabetes mellitus. In this study, the authors have attempted to relate various changes in genes expression or protein activity to the high-carbohydrate and high-MSG-induced diabetes. The authors have also tried to relate several genes that contribute to pathophysiology of T2D and proposed several ideas of genes as markers and target for curing people with T2D. These are done by investigating altered activities of various genes that cause or are caused by diabetes. These genes are selected based on their roles in pathophysiology of T2D.


Cranioplasty after Craniectomy in a Pediatric Population: Single-Center Experience from a Developing Country.
Cranioplasty is a frequently performed procedure in neurosurgery. The pediatric population for this procedure is distinct from the adult one because of the growing skulls and thinner bones of the calvarium. A paucity of data on the outcomes of this procedure in the pediatric population has been identified repeatedly. We conducted a retrospective cohort study to investigate the outcomes in a pediatric population that underwent cranioplasty after craniectomy at our institute in a developing-world country. Our cohort showed no association of complication rate or cosmetic outcomes with the timing of cranioplasty, area of skull defect, type of implant used, or method of storage.


An open-label, long-term evaluation of the safety, efficacy and tolerability of avanafil in male patients with mild to severe erectile dysfunction.
Determine the long-term efficacy, safety and tolerability of avanafil, a highly specific, rapidly absorbed phosphodiesterase type 5 inhibitor in male patients with mild to severe erectile dysfunction (ED), with or without diabetes. This was a 52-week, open-label extension of two 12-week, randomised, placebo-controlled, phase 3 trials. Patients were assigned to avanafil 100 mg, but could request 200 mg (for increased efficacy; '100/200-mg' group) or 50 mg (for improved tolerability). Primary end points included percentage of sexual attempts ending in successful vaginal penetration [Sexual Encounter Profile 2 (SEP2)] and intercourse (SEP3) and erectile function domain score per the International Index of Erectile Function (IIEF-EF). Some 712 patients enrolled; 686 were included in the intent to treat population and contributed to the data. All primary end points showed sustained improvement. SEP2 and SEP3 success rates improved from 44% to 83% and from 13% to 68% (100-mg group) and from 43% to 79% and from 11% to 66% (100/200-mg group), respectively. Mean IIEF-EF domain scores improved from 13.6 to 22.2 (100-mg group) and from 11.9 to 22.7 (100/200-mg group). Avanafil was effective in some patients ≤ 15 min and > 6 h postdose. Sixty-five per cent (112/172) of 'nonresponders' to avanafil 100 mg responded to the 200-mg dose. The most common (≥ 2%) treatment-emergent adverse events were headache, flushing, nasopharyngitis and nasal congestion; < 3% of patients discontinued therapy because of adverse events. The long-term tolerability and improvement in sexual function, coupled with rapid onset, suggest that avanafil is well suited for the on-demand treatment of ED.


Sexual conflict promotes speciation in insects.
Speciation rates among extant lineages of organisms vary extensively, but our understanding of the causes of this variation and, therefore, the processes of speciation is still remarkably incomplete. Both theoretical and empirical studies have indicated that sexual selection is important in speciation, but earlier discussions have focused almost exclusively on the potential role of female mate choice. Recent findings of postmating reproductive conflicts of interest between the sexes suggest a quite different route to speciation. Such conflicts may lead to perpetual antagonistic coevolution between males and females and may thus generate rapid evolutionary divergence of traits involved in reproduction. Here, we assess this hypothesis by contrasting pairs of related groups of insect species differing in the opportunity for postmating sexual conflict. Groups where females mate with many males exhibited speciation rates four times as high as in related groups where females mate only once. Our results not only highlight the general importance of postmating sexual selection in speciation, but also support the recent suggestion that sexual conflict is a key engine of speciation.


Lesions of the enamel organ of developing dog teeth following experimental inoculation of gnotobiotic puppies with canine distemper virus.
Ten 7-day-old gnotobiotic Beagle puppies were inoculated intraperitoneally with virulent canine distemper virus (R252-CDV). The dogs were killed and perfused with paraformaldehyde/glutaraldehyde from eight to 36 days after inoculation. The developing teeth of the mandibles were examined by light microscopy, and the teeth from three dogs were examined by electron microscopy. Necrosis of individual cells in the stratum intermedium of the developing tooth was the first change, detectable at day 9 post-inoculation. At day 16 post-inoculation, there was disorganization of the ameloblasts. In the stratum intermedium, multinucleate giant cells and large eosinophilic cytoplasmic viral inclusions were prominent. Ultrastructurally, these inclusions consisted of clusters of tubular aggregates typical of canine distemper virus nucleocapsids. At 28 to 36 days post-inoculation, the changes were seen in the reduced enamel epithelium. Multinucleate cells were seen, but no inclusions. Some necrotic cells were seen. In these teeth, ameloblastic cells of the root were morphologically normal. Our results suggest that distemper virus affects developing teeth by direct infection of the enamel organ.


Evaluation of Chronic Pain Using Magnetic Resonance (MR) Neuroimaging Approaches: What the Clinician Needs to Know.
Numerous neuroimaging techniques have been recently used to investigate central mechanisms involved in pain perception and to examine morphological and functional brain alterations associated with chronic pain. Compared to self-reporting approaches, objective imaging techniques are expected to potentially lead to better pain assessment and guide management. This comprehensive scoping review aims to identify recent magnetic resonance imaging (MRI) approaches that have been used to characterize the brain of chronic pain subjects, using structural, chemical and functional MRI techniques. A systematic search and review of the literature was conducted and the resultant studies were critically examined for relevance. MRI neuroimaging of various chronic pain conditions were summarized. We classified the collected studies into: structural brain alterations, VBM (voxel based morphology) examination of structural changes, DTI, changes in brain chemistry, functional and blood flow brain alterations. From our clinical experience, we have noted that most clinicians are not aware of the capabilities of advanced MRI methods in assessing cortical manifestations of chronic pain. In addition, many clinicians are not aware of the cortical alterations present in individuals with chronic pain. This comprehensive scoping review thus sets out to first summarize MRI neuroimaging techniques that are available in the current literature to examine chronic pain. We then identify cortical MR approaches that have been able to reliably predict transition from acute to chronic pain. Finally, we summarize MRI neuroimaging techniques that have been used to track treatment response of individuals with chronic pain.


The Schizophrenia Cognition Rating Scale: an interview-based assessment and its relationship to cognition, real-world functioning, and functional capacity.
Interview-based measures of cognition may serve as potential coprimary measures in clinical trials of cognitive-enhancing drugs for schizophrenia. However, there is no such valid scale available. Interviews of patients and their clinicians are not valid in that they are unrelated to patients' levels of cognitive impairment as assessed by cognitive performance tests. This study describes the reliability and validity of a new interview-based assessment of cognition, the Schizophrenia Cognition Rating Scale (SCoRS), that involves interviews with patients and informants. Sixty patients with schizophrenia were assessed with the SCoRS and three potential validators of an interview-based measure of cognition: cognitive performance, as measured by the Brief Assessment of Cognition in Schizophrenia (BACS); real-world functioning, as measured by the Independent Living Skills Inventory; and functional capacity, as measured by the University of California, San Diego, Performance-Based Skills Assessment (UPSA). The SCoRS global ratings were significantly correlated with composite scores of cognitive performance and functional capacity and with ratings of real-world functioning. Multiple regression analyses suggested that SCoRS global ratings predicted unique variance in real-world functioning beyond that predicted by the performance measures. An interview-based measure of cognition that included informant reports was related to cognitive performance as well as real-world functioning. Interview-based measures of cognition, such as the SCoRS, may be valid coprimary measures for clinical trials assessing cognitive change and may also aid clinicians desiring to assess patients' level of cognitive impairment.


Evolution and predictive value of IgE responses toward a comprehensive panel of house dust mite allergens during the first 2 decades of life.
The evolution of the IgE response to the numerous allergen molecules of Dermatophagoides pteronyssinus is still unknown. We sought to characterize the evolutionary patterns of the IgE response to 12 molecules of D pteronyssinus from birth to adulthood and to investigate their determinants and clinical relevance. We investigated the clinical data and sera of 722 participants in the German Multicenter Allergy Study, a birth cohort started in 1990. Diagnoses of current allergic rhinitis (AR) related to mite allergy and asthma were based on yearly interviews at the ages of 1 to 13 years and 20 years. IgE to the extract and 12 molecules of D pteronyssinus were tested by means of ImmunoCAP and microarray technology, respectively, in sera collected at ages 1, 2, 3, 5, 6, 7, 10, 13, and 20 years. Exposure to mites at age 6 and 18 months was assessed by measuring Der p 1 weight/weight concentration in house dust. One hundred ninety-one (26.5%) of 722 participants ever had IgE to D pteronyssinus extract (≥0.35 kUA/L). At age 20 years, their IgE recognized most frequently Der p 2, Der p 1, and Der p 23 (group A molecules; prevalence, >40%), followed by Der p 5, Der p 7, Der p 4, and Der p 21 (group B molecules; prevalence, 15% to 30%) and Der p 11, Der p 18, clone 16, Der p 14, and Der p 15 (group C molecules; prevalence, <10%). IgE sensitization started almost invariably with group A molecules and expanded sequentially first to group B and finally to group C molecules. Early IgE sensitization onset, parental hay fever, and higher exposure to mites were associated with a broader polymolecular IgE sensitization pattern. Participants reaching the broadest IgE sensitization stage (ie, ABC) had significantly higher risk of mite-related AR and asthma than unsensitized participants. IgE to Der p 1 or Der p 23 at age 5 years or less predicted asthma at school age. Parental hay fever and early exposure to D pteronyssinus allergens promote IgE polysensitization to several D pteronyssinus molecules, which in turn predicts current mite-related AR and current/future asthma. These results might inspire predictive algorithms and prevention strategies against the progression of IgE sensitization to mites toward AR and asthma.


Openness to experience and activity engagement facilitate the maintenance of verbal ability in older adults.
The current study used data from the Aberdeen Birth Cohort, 1936, to investigate the hypothesis that the positive effects of the personality trait Openness on cognitive ability are mediated by activity levels. Results of latent growth modeling analysis revealed that higher Openness predicted better reading ability, inductive reasoning, and memory performance across three testing occasions when participants were aged 64-68 years. Higher Openness predicted higher activity levels, and higher activity levels in turn predicted higher reading ability, but not higher performance on measures of inductive reasoning, memory, and speed of processing. Overall, Openness and activity engagement appear related to preserved higher cognitive ability in older adults, with Openness having a direct effect on marker tests of fluid ability and with the combined influence of Openness and activity being particularly important for marker tests of crystallized intelligence.


Associations between violent video gaming, empathic concern, and prosocial behavior toward strangers, friends, and family members.
Exposure to media violence, including violent video gaming, can have a cognitive desensitization effect, lowering empathic concern for others in need. Since emerging adulthood offers increased opportunities to volunteer, strengthen relationships, and initiate new relationships, decreases in empathic concern and prosocial behavior may prove inhibitive to optimal development during this time. For these reasons, the current study investigated associations between violent video gaming, empathic responding, and prosocial behavior enacted toward strangers, friends, and family members. Participants consisted of 780 emerging adults (M age = 19.60, SD = 1.86, range = 18–29, 69% female, 69% Caucasian) from four universities in the United States. Results showed small to moderate effects between playing violent video gaming and lowered empathic concern for both males and females. In addition, lowered empathic concern partially mediated the pathways between violent video gaming and prosocial behavior toward all three targets (at the level of a trend for females), but was most strongly associated with lower prosocial behavior toward strangers. Discussion highlights how violent video gaming is associated with lower levels of prosocial behavior through the mechanism of decreased empathic concern, how this association can affect prosocial behavior differently across target, and finally what implications this might have for development during emerging adulthood.


Distinct chromophore-protein environments enable asymmetric activation of a bacteriophytochrome-activated diguanylate cyclase.
Sensing of red and far-red light by bacteriophytochromes involves intricate interactions between their bilin chromophore and the protein environment. The light-triggered rearrangements of the cofactor configuration and eventually the protein conformation enable bacteriophytochromes to interact with various protein effector domains for biological modulation of diverse physiological functions. Excitation of the holoproteins by red or far-red light promotes the photoconversion to their far-red light-absorbing Pfr state or the red light-absorbing Pr state, respectively. Because prototypical bacteriophytochromes have a parallel dimer architecture, it is generally assumed that symmetric activation with two Pfr state protomers constitutes the signaling-active species. However, the bacteriophytochrome from Idiomarina species A28L (IsPadC) has recently been reported to enable long-range signal transduction also in asymmetric dimers containing only one Pfr protomer. By combining crystallography, hydrogen-deuterium exchange coupled to MS, and vibrational spectroscopy, we show here that Pfr of IsPadC is in equilibrium with an intermediate "Pfr-like" state that combines features of Pfr and Meta-R states observed in other bacteriophytochromes. We also show that structural rearrangements in the N-terminal segment (NTS) can stabilize this Pfr-like state and that the PHY-tongue conformation of IsPadC is partially uncoupled from the initial changes in the NTS. This uncoupling enables structural asymmetry of the overall homodimeric assembly and allows signal transduction to the covalently linked physiological diguanylate cyclase output module in which asymmetry might play a role in the enzyme-catalyzed reaction. The functional differences to other phytochrome systems identified here highlight opportunities for using additional red-light sensors in artificial sensor-effector systems.


Contraception and abortion in a low-fertility setting: the role of seasonal migration.
Seasonal labor migration is common among men in many former Soviet republics. Little research has examined contraceptive use and induced abortion among women in such low-fertility, high-migration settings, according to husband's migration status. Combined data from 2,280 respondents of two surveys of married women aged 18-45 in rural Armenia-one conducted in 2005 and one in 2007-were used. Logistic regression analyses examined whether a husband's migration status was associated with his wife's current use of the pill or the IUD, or with the probability that she had had a pregnancy that ended in induced abortion. Additional analyses were conducted to determine whether relationships were moderated by household wealth. Women with a migrant husband were less likely than those with a nonmigrant husband to be currently using the pill or the IUD (odds ratio, 0.6); with increased household wealth, the likelihood of method use increased among women with a nonmigrant husband, but decreased slightly among women with a migrant husband. Overall, the probability that a pregnancy ended in abortion did not differ by migration status; however, the likelihood of abortion increased with wealth among women married to a nonmigrant, but not among those married to a migrant. Despite their husband's absence, women married to a migrant may have an unwanted pregnancy rate similar to that of women married to a nonmigrant. Improved access to modern contraceptive methods is likely to be positively associated with contraceptive use among women with a nonmigrant husband, but not among those with a migrant husband.


[Swallowing after non-surgical treatment (radiation therapy / radiochemotherapy protocol) of laryngeal cancer].
Radiation therapy and radiochemotherapy protocols can cause swallowing difficulties. To evaluate swallowing in patients undergoing radiation therapy and radiochemotherapy protocol only for the treatment of laryngeal tumors. A prospective study of 20 patients, with a mean age of 62 years, at the end of oncological therapy. Six patients (30%) underwent radiation therapy, and 14 patients (70%) underwent combined therapy. The mean time between treatment and an evaluation of swallowing was 8.5 months. Videofluoroscopy was done to assess the preparatory, oral and pharyngeal phases of swallowing. All patients had only an oral diet. Normal swallowing was present in only 25% of patients. The swallowing videofluoroscopic examination identified the following changes: bolus formation (85%), bolus ejection (60%), oral cavity stasis (55%), changes in the onset of the pharyngeal phase (100%), decreased laryngeal elevation (65%), and hypopharyngeal stasis (80%). Laryngeal penetration was observed in 25% of the cases; 40% presented tracheal aspiration. The grade of penetration/aspiration was mild in 60% of cases. Aspiration was silent in 35% of patients. Although 75% of patients had dysphagia, only 25% complained of swallowing difficulties. Patients with laryngeal cancer that underwent radiation therapy/combined treatment can present changes in all swallowing phases, or may be asymptomatic.


Dexmedetomidine-Induced Neuroapoptosis Is Dependent on Its Cumulative Dose.
Dexmedetomidine (DEX) has inherent neuroprotective properties that have been attributed to the activation of prosurvival kinases. However, the impact of supraclinical doses of DEX on neuroapoptosis and neuronal viability has not been determined. Rat pups and primary neuronal cells were treated with DEX or ketamine (KET) alone or in combination. Neuroapoptosis was measured by cleaved-caspase-3 expression and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining in brain sections. Expression of prosurvival kinases was measured by Western blot. We measured the impact of DEX with and without α1-adrenergic receptor blockade on the viability of primary neuronal cell cultures. Increasing the cumulative dose of DEX resulted in elevated levels of neuroapoptosis in vivo. Low doses increased, whereas high dose decreased phosphorylation of the prosurvival kinases. KET alone and in combination with DEX produced a greater degree of apoptosis and reductions in expression of these protein kinases than DEX alone. Increasing concentrations of DEX decreased, while coadministration of an α1-adrenergic receptor blocker preserved neuronal viability in vitro. Although DEX is neuroprotective at clinical doses, high cumulative doses and concentrations induce neuroapoptosis, in vivo and in vitro, respectively. Because the current dosing schedules used in humans yield plasma levels that are substantially below concentrations that induce neurotoxicity, low-dose DEX should not be neurotoxic and has the potential to be a neuroprotective adjuvant.


Effect of supragingival plaque control in smokers and never-smokers: 6-month evaluation of patients with periodontitis.
The effect of supragingival plaque control on clinical signs of periodontitis is controversial, particularly when smoking habits are considered. This study evaluated the clinical effects of supragingival plaque control on clinical signs of periodontitis in smokers and never-smokers. The following data were collected for 25 never-smokers and 25 smokers at baseline and 30, 90, and 180 days: visible plaque index (VPI), gingival bleeding index (GBI), bleeding on probing (BOP), periodontal probing depth (PD), and clinical attachment loss (CAL). After baseline examinations, supragingival scaling was performed. Oral hygiene practices were reinforced and reevaluated weekly during the experimental period. Linear models adjusted for clustering of observations within individuals were used for statistical analysis. Reductions in VPI were significant for both groups, with no intergroup differences. GBI at baseline was similar between groups, and at 30, 90, and 180 days, smokers had a lower GBI than never-smokers. Significant reductions were observed in PD for shallow (1 to 3 mm), moderate (4 to 5 mm), and deep sites (> or = 6 mm) in both groups. CAL was significantly greater in smokers throughout the study, but gains in attachment were similar for both groups (0.71 to 1.00 mm). BOP reductions were similar in both groups. Supragingival plaque control resulted in significant changes in clinical parameters associated with gingivitis and periodontitis. Smoking did not affect results, regardless of initial PD.


Doing better by getting worse: posthypnotic amnesia improves random number generation.
Although forgetting is often regarded as a deficit that we need to control to optimize cognitive functioning, it can have beneficial effects in a number of contexts. We examined whether disrupting memory for previous numerical responses would attenuate repetition avoidance (the tendency to avoid repeating the same number) during random number generation and thereby improve the randomness of responses. Low suggestible and low dissociative and high dissociative highly suggestible individuals completed a random number generation task in a control condition, following a posthypnotic amnesia suggestion to forget previous numerical responses, and in a second control condition following the cancellation of the suggestion. High dissociative highly suggestible participants displayed a selective increase in repetitions during posthypnotic amnesia, with equivalent repetition frequency to a random system, whereas the other two groups exhibited repetition avoidance across conditions. Our results demonstrate that temporarily disrupting memory for previous numerical responses improves random number generation.


The "patholog" of the genes expression profile, a new tool in defining, evaluating, and classifying genetic diseases.
The paper introduces the pre-Hilbert space of standard gene expression (SSGE) based on the normal variability in gene profiles, as revealed by the "spotting" microarray technique. In this space, every point represents a possible gene profile and every continuous curve a possible genetic evolution. The gene "patholog" is defined as the Euclidean distance separating the representative point of the gene profile from the unit, 0-centered hyperball. It is the most general quantification of the alteration in digital genes expression, suitable to define, evaluate, and classify the genetic diseases. Our mathematical model and the afferent computer package allow the researcher to identify the patterns of various genetic afflictions in the SSGE. A publicly available database will be opened where every interested experimentalist could introduce his/her results and process them according to our procedure.


Cross-talk between 10-gingerol and its anti-cancerous potential: a recent update.
Since time immortal, ginger, as an ancient herb, has been used throughout the world in foods and beverages due to its typical strong and pungent flavor. Besides its use as a spice, it also serves as an excellent source of several bioactive phenolics, including nonvolatile pungent compounds, such as gingerols, paradols, shogaols, and gingerones. Gingerols constitute key ingredients in fresh ginger, with the most abundant being 6-gingerol (6-G), 8-gingerol (8-G), and 10-gingerol (10-G). Many studies have investigated the various valuable pharmacological properties of these ingredients and experimentally verified the mechanistic aspects of their health effects; however, to date, most research on the anti-cancerous activities of gingerols have focused largely on 6-G. Thus, the present article deals with the number of recent studies that have indicated and highlighted the role of 10-G with respect to its cancer prevention attributes in particular and its anti-inflammatory, anti-oxidant, anti-microbial, and gastrointestinal tract protective potential in general. The purpose of this review is to provide an overview of all the experimentally validated health benefits of 10-G for nutraceutical applications. The various findings have warranted the further investigation of 10-G and its possible use in various cancer treatments as well as its promising role as a chemo-preventive agent.


Na+/Ca2+ exchanger is a determinant of excitation-contraction coupling in human embryonic stem cell-derived ventricular cardiomyocytes.
In adult cardiomyocytes (CMs), the Na(+)/Ca(2+) exchanger (NCX) is a well-defined determinant of Ca(2+) homeostasis. Developmentally, global NCX knockout in mice leads to abnormal myofibrillar organization, electrical defects, and early embryonic death. Little is known about the expression and function of NCX in human heart development. Self-renewable, pluripotent human embryonic stem cells (hESCs) can serve as an excellent experimental model. However, hESC-derived CMs are highly heterogeneous. A stably lentivirus-transduced hESC line (MLC2v-dsRed) was generated to express dsRed under the transcriptional control of the ventricular-restricted myosin light chain-2v (MLC2v) promoter. Electrophysiologically, dsRed+ cells differentiated from MLC2vdsRed hESCs displayed ventricular action potentials (AP), exclusively. Neither atrial nor pacemaker APs were observed. While I(Ca-L), I(f), and I(Kr) were robustly expressed, I(Ks) and I(K1) were absent in dsRed+ ventricular hESCCMs. Upon differentiation (7+40 to +90 days), the basal [Ca(2+)](i), Ca(2+) transient amplitude, maximum upstroke, and decay velocities significantly increased (P < 0.05). The I(Ca-L) antagonizer nifedipine (1 microM) decreased the Ca(2+) transient amplitude (to approximately 30%) and slowed the kinetics (by approximately 2-fold), but Ca(2+) transients could still be elicited even after complete ICa-L blockade, suggesting the presence of additional Ca(2+) influx(es). Indeed, Ni(2+)-sensitive INCX could be recorded in 7+40- and +90-day dsRed+ hESC-CMs, and its densities increased from -1.2 +/- 0.6 pA/pF at -120 mV and 3.6 +/- 1.0 pA/pF at 60 mV by 6- and 2-folds, respectively. With higher [Ca(2+)](i), 7+90-day ventricular hESC-CMs spontaneously but irregularly fired transients upon a single stimulus under an external Na(+)-free condition; however, without extracellular Na(+), nifedipine could completely inhibit Ca(2+) transients. We conclude that I(NCX) is functionally expressed in developing ventricular hESC-CMs and contributes to their excitation-contraction coupling.


Targeting tumor-initiating cancer cells with dCD133KDEL shows impressive tumor reductions in a xenotransplant model of human head and neck cancer.
A novel anticancer agent was constructed by fusing a gene encoding the scFV that targets both glycosylated and unglycosylated forms of CD133 to a gene fragment encoding deimmunized PE38KDEL. The resulting fusion protein, dCD133KDEL, was studied to determine its ability to bind and kill tumor-initiating cells in vitro and in vivo. The anti-CD133 scFV selectively bound HEK293 cells transfected with the CD133 receptor gene. Time course viability studies showed that dCD133KDEL selectively inhibited NA-SCC and UMSCC-11B, 2 head and neck squamous cell carcinomas that contain a CD133 expressing subpopulation. Importantly, the drug did not inhibit the viability of hematopoietic lineages measured by long-term culture-initiating cell and colony-forming assays from sorted human CD34+ progenitor cells. In addition to in vitro studies, in vivo tumor initiation experiments confirmed that CD133-sorted cells implanted into the flanks of nude mice grew faster and larger than unsorted cells. In contrast, cells that were pretreated with dCD133KDEL before implantation showed the slowest and lowest incidence of tumors. Furthermore, UMSCC-11B-luc tumors treated with multiple intratumoral injections of dCD133KDEL showed marked growth inhibition, leading to complete degradation of the tumors that was not observed with an irrelevant control-targeted toxin. Experiments in immunocompetent mice showed that toxin deimmunization resulted in a 90% reduction in circulating antitoxin levels. These studies show that dCD133KDEL is a novel anticancer agent effective at inhibiting cell proliferation, tumor initiation, and eliminating established tumors by targeting the CD133 subpopulation. This agent shows significant promise for potential development as a clinically useful therapy.


Multi-port polarization-independent optical quasi-circulators by using a pair of holographic spatial- and polarization- modules.
In this paper we proposed an alternative type of multi-port polarization-independent optical quasi-circulator by using a pair of holographic spatial- and polarization- modules. The prototype is fabricated and experimentally tested. In addition, the operating principles, the characteristics and the performances of this device are discussed. The merits of this design include polarization-independence, compactness, high isolation, low polarization mode dispersion, and easy fabrication. Furthermore, the number of ports can be scaled up easily.


Central and statistical data monitoring in the Clinical Randomisation of an Antifibrinolytic in Significant Haemorrhage (CRASH-2) trial.
Background The purpose of monitoring in clinical trials is to ensure the rights, safety, and well-being of trial patients and the accuracy of the trial data. In the Clinical Randomisation of an Antifibrinolytic in Significant Haemorrhage (CRASH-2) trial, which recruited over 20,000 adult trauma patients worldwide, the nature and extent of monitoring was based on a risk assessment undertaken before recruitment started. Purpose We report the methods used for central and statistical monitoring in the CRASH-2 trial and explain how central monitoring was used to target on-site investigations. Methods To ensure that trial participants met the inclusion criteria, we monitored event rates for the primary (death) and secondary outcomes (blood transfusion given). We monitored four quantitative variables (systolic blood pressure (SBP), heart rate (HR), respiratory rate, and capillary refill time) as indicators of the severity of bleeding. We used the coefficient of variation (CV) to identify sites with too much or too little variability. To ensure the accuracy of the data on side effects, we monitored thromboembolic events at each site. Sites with higher or lower than expected event rates were identified for further evaluation. Results A total of 274 sites recruited patients: 145 sites recruited ≥20; patients, and 52 sites recruited ≥100 patients. Sites with low case fatality and low blood transfusion rates were found to be including patients with relatively mild haemorrhage. One site with a high rate of thromboembolic events was found to be using clinical judgement alone. Measurements of SBP and HR varied by about one-fifth of their average value, and capillary refill time measurements varied by around one-third of their average; between-site variation was lowest for blood pressure. Limitations A comparison of mean and median CV indicated that the distributions are slightly skewed to the right. Our simple approach to calculating 95% confidence intervals for the CV may be improved by using a logarithmic transformation of CV for each variable. Conclusions Central and statistical monitoring of data can be used to monitor clinical trials, particularly large, pragmatic, international trials where 100% on-site monitoring is neither necessary nor cost-effective. In the CRASH-2 trial, re-education about trial protocol and the development of guidance helped resolve the issues identified during monitoring.