Temporal order of evolution of DNA replication systems inferred by comparison of cellular and viral DNA polymerases.
The core enzymes of the DNA replication systems show striking diversity among cellular life forms and more so among viruses. In particular, and counter-intuitively, given the central role of DNA in all cells and the mechanistic uniformity of replication, the core enzymes of the replication systems of bacteria and archaea (as well as eukaryotes) are unrelated or extremely distantly related. Viruses and plasmids, in addition, possess at least two unique DNA replication systems, namely, the protein-primed and rolling circle modalities of replication. This unexpected diversity makes the origin and evolution of DNA replication systems a particularly challenging and intriguing problem in evolutionary biology. I propose a specific succession for the emergence of different DNA replication systems, drawing argument from the differences in their representation among viruses and other selfish replicating elements. In a striking pattern, the DNA replication systems of viruses infecting bacteria and eukaryotes are dominated by the archaeal-type B-family DNA polymerase (PolB) whereas the bacterial replicative DNA polymerase (PolC) is present only in a handful of bacteriophage genomes. There is no apparent mechanistic impediment to the involvement of the bacterial-type replication machinery in viral DNA replication. Therefore, I hypothesize that the observed, markedly unequal distribution of the replicative DNA polymerases among the known cellular and viral replication systems has a historical explanation. I propose that, among the two types of DNA replication machineries that are found in extant life forms, the archaeal-type, PolB-based system evolved first and had already given rise to a variety of diverse viruses and other selfish elements before the advent of the bacterial, PolC-based machinery. Conceivably, at that stage of evolution, the niches for DNA-viral reproduction have been already filled with viruses replicating with the help of the archaeal system, and viruses with the bacterial system never took off. I further suggest that the two other systems of DNA replication, the rolling circle mechanism and the protein-primed mechanism, which are represented in diverse selfish elements, also evolved prior to the emergence of the bacterial replication system. This hypothesis is compatible with the distinct structural affinities of PolB, which has the palm-domain fold shared with reverse transcriptases and RNA-dependent RNA polymerases, and PolC that has a distinct, unrelated nucleotidyltransferase fold. I propose that PolB is a descendant of polymerases that were involved in the replication of genetic elements in the RNA-protein world, prior to the emergence of DNA replication. By contrast, PolC might have evolved from an ancient non-templated polymerase, e.g., polyA polymerase. The proposed temporal succession of the evolving DNA replication systems does not depend on the specific scenario adopted for the evolution of cells and viruses, i.e., whether viruses are derived from cells or virus-like elements are thought to originate from a primordial gene pool. However, arguments are presented in favor of the latter scenario as the most parsimonious explanation of the evolution of DNA replication systems. Comparative analysis of the diversity of genomic strategies and organizations of viruses and cellular life forms has the potential to open windows into the deep past of life's evolution, especially, with the regard to the origin of genome replication systems. When complemented with information on the evolution of the relevant protein folds, this comparative approach can yield credible scenarios for very early steps of evolution that otherwise appear to be out of reach. Eric Bapteste, Patrick Forterre, and Mark Ragan.


Metal Exposure during Early Pregnancy and Risk of Gestational Diabetes Mellitus: Mixture Effect and Mediation by Phospholipid Fatty Acids.
Despite existing studies exploring the association between metal exposure and gestational diabetes mellitus (GDM), most of them have focused on a single metal or a small mixture of metals. Our prospective work investigated the joint and independent effects of early gestational exposure to 17 essential and nonessential metals on the GDM risk and potential mediation by plasma phospholipid fatty acids (PLFAs) based on a nested case-control study established with 335 GDM cases and 670 randomly matched healthy controls. The Bayesian kernel machine regression (BKMR) and quantile g-computation analyses demonstrated a joint effect from metal co-exposure on GDM risk. BKMR with hierarchical variable selection indicated that the group of essential metals was more strongly associated with GDM than the group of nonessential metals with group posterior inclusion probabilities (PIPs) of 0.979 and 0.672, respectively. Cu (0.988) and Ga (0.570) had the largest conditional PIPs within each group. We also observed significant mediation effects of selected unsaturated PLFAs on Cu-GDM and Ga-GDM associations. KEGG enrichment analysis further revealed significant enrichment in the biosynthesis of unsaturated PLFAs. C18:1 n-7 exhibited the largest proportion of mediation in both associations (23.8 and 22.9%). Collectively, our work demonstrated the joint effect of early gestational metal exposure on GDM risk and identified Cu and Ga as the key species to the joint effect. The findings lay a solid ground for further validation through multicenter investigations and mechanism exploration via laboratory studies.


Ontogenetic shifts in fishes between vegetated and unvegetated tidepools: assessing the effect of physical structure on fish habitat selection.
The aim of this study of tidepool fishes was analyse variation in their use of intertidal habitats (rocky shore, mangrove and salt marsh). Specimens were collected during wet and dry periods from 18 tidepools in the three habitats. A total of 7690 specimens, belonging to 19 families and 30 species, was captured. The fish assemblage in rocky shore pools was clearly distinct from that of vegetated habitats (mangrove and salt marshes). The rocky shore fauna was dominated by permanent resident species, whereas pools in mangrove and salt marsh habitats were inhabited primarily by opportunistic and transient species. Habitat segregation by ontogenetic stage (e.g. smaller individuals in mangroves, intermediate size classes in salt marsh and sub-adults/adults on rocky shores) indicates age-related migration in response to the physical structure of these habitats and to the natural history of each fish species. These findings are important for the development of effective conservation and management plans for intertidal fishes.


Effects of interleukin-1 on bone turnover in normal mice.
Interleukin-1 (IL-1) is a potent stimulator of osteoclastic bone resorption in vitro and causes hypercalcemia and increased osteoclastic resorption when infused into normal mice for 72 h. However, its longer term or local effects on bone turnover in vivo are unknown. To study these, we injected IL-1 alpha once daily for 3 days into the sc tissue over the calvariae of normal mice and examined its effects on calvarial bone morphology during the subsequent 4 weeks using quantitative histomorphometry. Increased bone resorption inside the calvariae and elevated plasma calcium concentrations were present 24 h after the last IL-1 injection. These early systemic effects were not prevented by indomethacin. During the following 3-4 weeks most of the bone on the injected side of the calvariae was resorbed by osteoclasts and was subsequently replaced by increased amounts of new bone. These longer term local effects on bone turnover were prevented by indomethacin. However, indomethacin did not prevent the formation of new bone inside the calvariae at sites of resorption induced by IL-1 independent of prostaglandin production. These findings indicate that IL-1 stimulates bone turnover systemically, independent of prostaglandin production, and that it has profound long term local effects on bone turnover that are mediated through prostaglandins.


An insect-inspired model for visual binding II: functional analysis and visual attention.
We have developed a neural network model capable of performing visual binding inspired by neuronal circuitry in the optic glomeruli of flies: a brain area that lies just downstream of the optic lobes where early visual processing is performed. This visual binding model is able to detect objects in dynamic image sequences and bind together their respective characteristic visual features-such as color, motion, and orientation-by taking advantage of their common temporal fluctuations. Visual binding is represented in the form of an inhibitory weight matrix which learns over time which features originate from a given visual object. In the present work, we show that information represented implicitly in this weight matrix can be used to explicitly count the number of objects present in the visual image, to enumerate their specific visual characteristics, and even to create an enhanced image in which one particular object is emphasized over others, thus implementing a simple form of visual attention. Further, we present a detailed analysis which reveals the function and theoretical limitations of the visual binding network and in this context describe a novel network learning rule which is optimized for visual binding.


An Anion Conductance, the Essential Component of the Hydroxyl-Radical-Induced Ion Current in Plant Roots.
Oxidative stress signaling is essential for plant adaptation to hostile environments. Previous studies revealed the essentiality of hydroxyl radicals (HO•)-induced activation of massive K⁺ efflux and a smaller Ca2+ influx as an important component of plant adaptation to a broad range of abiotic stresses. Such activation would modify membrane potential making it more negative. Contrary to these expectations, here, we provide experimental evidence that HO• induces a strong depolarization, from -130 to -70 mV, which could only be explained by a substantial HO•-induced efflux of intracellular anions. Application of Gd3+ and NPPB, non-specific blockers of cation and anion conductance, respectively, reduced HO•-induced ion fluxes instantaneously, implying a direct block of the dual conductance. The selectivity of an early instantaneous HO•-induced whole cell current fluctuated from more anionic to more cationic and vice versa, developing a higher cation selectivity at later times. The parallel electroneutral efflux of K⁺ and anions should underlie a substantial leak of the cellular electrolyte, which may affect the cell's turgor and metabolic status. The physiological implications of these findings are discussed in the context of cell fate determination, and ROS and cytosolic K⁺ signaling.


What factors control superficial lava dome explosivity?
Dome-forming eruption is a frequent eruptive style and a major hazard on numerous volcanoes worldwide. Lava domes are built by slow extrusion of degassed, viscous magma and may be destroyed by gravitational collapse or explosion. The triggering of lava dome explosions is poorly understood: here we propose a new model of superficial lava-dome explosivity based upon a textural and geochemical study (vesicularity, microcrystallinity, cristobalite distribution, residual water contents, crystal transit times) of clasts produced by key eruptions. Superficial explosion of a growing lava dome may be promoted through porosity reduction caused by both vesicle flattening due to gas escape and syn-eruptive cristobalite precipitation. Both processes generate an impermeable and rigid carapace allowing overpressurisation of the inner parts of the lava dome by the rapid input of vesiculated magma batches. The relative thickness of the cristobalite-rich carapace is an inverse function of the external lava dome surface area. Explosive activity is thus more likely to occur at the onset of lava dome extrusion, in agreement with observations, as the likelihood of superficial lava dome explosions depends inversely on lava dome volume. This new result is of interest for the whole volcanological community and for risk management.


Afterdepolarizations promote the transition from ventricular tachycardia to fibrillation in a three-dimensional model of cardiac tissue.
Recent experimental results regarding the action potential duration restitution curve have explained the transition from ventricular tachycardia (VT) to fibrillation (VF) in terms of spiral wave (SW) meandering and breakup. However, it remains unclear whether VF always has a steep restitution curve. The present study was designed to test the hypothesis that afterdepolarizations occur at excitable gaps during VF and affect the SW dynamics, even if the restitution curve is gentle. Homogeneous and isotropic 3-dimensional tissue was simulated with a LRd model. Because of the gentle restitution curve, it was not expected that SW instabilities would occur in this condition. In the tissue, a stationary SW reentry was initially observed; however, afterdepolarizations erupted from the excitable gap near the SW tip, and the SW then meandered widely. Following that, afterdepolarizations erupted far from the SW tip, resulting in SW breakup. In this manner, the wave dynamics degenerated into a chaotic state within a few seconds. Furthermore, not only triggered activity but also subthreshold afterdepolarizations were found to cause SW instabilities. These results suggest that afterdepolarizations may play an important role in the transition to VF and that the mechanism is independent of restitution properties.


Decreased Resting-State Activity in the Precuneus Is Associated With Depressive Episodes in Recurrent Depression.
To investigate alterations in resting-state spontaneous brain activity in patients with major depressive disorder (MDD) experiencing multiple episodes. Between May 2007 and September 2014, 24 recurrent and 22 remitted patients diagnosed with MDD with the Structured Clinical Interview for DSM-IV Axis I Disorders (SCID-I), and 69 healthy controls matched for age, sex, and educational level participated in this study. Among them, 1 healthy control was excluded due to excessive head motion. The fractional amplitude of low-frequency fluctuation (fALFF) was assessed for all recruited subjects during the completion of resting-state functional magnetic resonance imaging. Relationships between fALFF and clinical measurements, including number of depressive episodes and illness duration, were examined. Compared to patients with remitted MDD and to healthy controls, patients with recurrent MDD exhibited decreased fALFF in the right posterior insula and right precuneus and increased fALFF in the left ventral anterior cingulate cortex. Decreased fALFF in the right precuneus and increased fALFF in the right middle insula were correlated with the number of depressive episodes in the recurrent MDD groups (r = -0.75, P < .01 and r = 0.78, P < .01, respectively) and remitted MDD groups (r = -0.63, P < .01 and r = 0.41, P = .03, respectively). In addition to regions in the default mode network (DMN) and salience network, the altered resting-state activity in the middle temporal and visual cortices was also identified. Altered resting-state activity was observed across several neural networks in patients with recurrent MDD. Consistent with the emerging theory that altered DMN activity is a risk factor for depression relapses, the association between reduced fALFF in the right precuneus and number of depressive episodes supports the role of the DMN in the pathology of recurrent depression.


Insulin-like growth factor I in fetal serum obtained by cordocentesis is correlated with intrauterine growth retardation.
We examined whether insulin-like growth factor-I (IGF-I) and one of its binding proteins (IGFBP-1) in fetal serum obtained by cordocentesis is correlated with intrauterine growth retardation (IUGR) and weight estimation by ultrasound. Cordocentesis sera from 27 fetuses suspected of having IUGR were analysed for IGF-I and IGFBP-1 by radioimmunoassay. The results showed that IGF-I concentrations were correlated significantly with birth weight (P < 0.001) and placenta weight (P < 0.05). Mean fetal concentrations of IGF-I were 38 +/- 18 microg/l. In patients (n = 11) with a weight deviation at delivery <-33%, IGF-I concentrations were 24.1 +/- 13.2 microg/l. IGFBP-1 was inversely correlated with birth weight (P < 0.006) and concentrations of IGF-I. Mean plasma concentrations of IGFBP-1 were 234.2 +/- 161.4 microg/l. Furthermore, IGF-I concentrations were correlated with the weight deviation estimated by ultrasonography at the time of cordocentesis (P < 0.007), as well as with the weight deviation at delivery (P < 0.0001). The actual weight deviation at delivery was correlated more strongly with fetal IGF-I concentrations than with the estimated weight deviation at cordocentesis. The lowest concentrations of IGF-I were found in patients with a weight deviation <-33%. Very low concentrations of IGF-I are thus associated with IUGR, indicating that IGF-I measured in fetal serum may increase the predictive value of ultrasonographic weight estimation.


Association Between Alcohol Use Among College Students and Alcohol Outlet Proximity and Densities.
Alcohol consumption is common on college campuses and is associated with negative consequences. Factors associated with availability of alcohol are not completely understood. To describe how proximity and density of alcohol outlets are associated with any drinking and binge drinking in students at the University of Wisconsin-Madison. Participants were full-time students enrolled in the Young Adults Eating and Active for Health, a multisite, randomized intervention that assessed a variety of health behaviors. Geographic information systems were used to calculate proximity and enumerate alcohol outlet densities. Participants were categorized as "drinkers" or "nondrinkers" based on self-reported alcohol consumption. Binge drinking was categorized as "non-binge drinker," "frequent binge drinker," and "excessive binge drinker." Analysis included regression, t tests, and chi-square tests. RESULTS. Among the 166 participants, 126 (76%) were drinkers. Among drinkers, 80 (63%) were either frequent or excessive binge drinkers. Drinkers lived closer to an alcohol outlet than non-drinkers (0.18 +/- 0.15 vs. 0.61 +/- 1.59 miles, respectively, P=0.005). Within a 1-mile walking radius, there were 47% more establishments for drinkers (153 +/- 47 compared to 104 +/- 55 outlets for nondrinkers, P<0.0001). At distances of 0.10-0.25 and 0.25-0.50 miles, twice as many outlets were available to drinkers (19 +/- 19 and 43 +/- 25, respectively) compared to nondrinkers (7 +/- 11 and 20 +/- 22, respectively), P<0.001. Proximity and density were hot associated with binge drinking frequency. Drinkers lived closer to alcohol outlets and had significantly more outlets available at a distance of up to 1 mile. Municipal and college administrators could consider limiting alcohol license distributions in municipalities with high alcohol consumption.


Improvement of phytochemical and quality characteristics of Dracocephalum kotschyi by drying methods.
This experiment was conducted to evaluate the effects of different drying methods on drying parameters and qualitative characteristics of Dracocephalum kotschyi in a completely randomized design with three replications. Treatments included shade drying as control, sun drying, cabinet drying (CD at 50 and 60°C), refractance window drying (RWD), infrared drying (IRD) at 200 and 300 W, and combination of RWD+ IRD at 200 and 300 W. According to the results, IRD, RWD, and RWD+ IRD effectively maintained valuable secondary metabolites compared to the conventional drying methods. The maximum total phenol content (2.7 and 2.66 mg GAE/g dry weight), total flavonoid content (2.26 and 2.33 mg QE/g dry weight), antioxidant activity (79% and 78.33%), and essential oil content (0.65% and 0.76%) were obtained from plants dried by RWD and IRD. Samples dried by RWD, IRD, and RWD+ IRD had high color quality, acceptable green color, and less browning. Also, RWD and IRD methods effectively reduced microbial contamination of dried plants compared to the control and other methods. The minimum aerobic mesophiles, mold, yeast, and coliforms were observed at 3.11, 0, and 1.47 log CFU/g in IRD 300 W and 3.17, 1, and 1.30 log CFU/g in RWD. D. kotschyi dried at CD 50°C had the maximum microbial contamination. Generally, according to the obtained results, RWD and IRD methods are suggested for drying of D. kotschyi and similar herbs due to shortening the drying time, preserving and improving the quality properties of dried plants.


Phage therapy administered noninvasively could be effective in thin tubes subject to episodic flow despite washout: a simulation study.
Bacteriophages (phages) have been proposed as candidates for the treatment of bacterial infections in light of emerging antibiotic-resistant microorganisms. Bacterial growth within thin tubes is a particular concern, such as in urinary tract infections and colonization of catheters. However, it is not clear whether phage administration to the urinary tract or in catheters could be effective in the context of flow to the outside (i.e. voiding or saline flush). Here, we adapt a previous model of phage infection to a thin tube geometry mimicking the spatial organization of the urinary tract, including bacterial motility and episodic flow during which phages are washed out of the system. We show that density-dependent dynamics permit propagation of the phage infection and that washout has little effect on the timing of bacterial clearance. In addition, instillation of phage at the bottom ~0.1 mm of the tract is effective in our computational model, suggesting that therapeutic phage introduced non-invasively could be efficacious in such situations.


Maintenance therapy in COPD: time to phase out ICS and switch to the new LAMA/LABA inhalers?
Long-term maintenance therapy for COPD is evolving rapidly. Dual bronchodilation with new long-acting muscarinic antagonist and long-acting beta-agonist (LAMA/LABA) fixed dose combination inhalers were introduced over the past 2 years. In clinical trials, these inhalers significantly improved lung function (trough forced expiratory volume in 1 second), patient-reported outcomes, and quality of life measures compared with placebo, their respective monocomponents, and tiotropium. The recorded adverse events of these new inhalers were also similar to those of their monocomponents or placebo. There are concerns regarding long-term complications (weight gain, osteoporosis, cataract) and increased risk of community-acquired pneumonia with the use of inhaled corticosteroids (ICS). The new LAMA/LABA inhalers could potentially reduce the use of ICS as part and parcel of maintenance therapy in COPD. Recent studies compared these LAMA/LABA inhalers with ICS/LABA combination inhalers in moderate-to-severe COPD. The results are promising and favor the LAMA/LABA inhalers, especially in the longer duration trials. Furthermore, there is a clearer picture emerging as to the subgroup of COPD patients who may be able to successfully switch from their current ICS/LABA therapy to these new LAMA/LABA inhalers.


Developing an information prescription service.
Availability and accessibility of high-quality information is inconsistent nationally and improving information is central to Department of Health policy. This article outlines an information prescription (IP) pilot. Using a quality-improvement cycle, feedback was gathered from stakeholders to develop an IP service for people with Parkinson's disease and their relatives/carers. Generally, service users were satisfied with the information and felt it enabled them to better understand, manage and be in control of their condition. Despite initial concerns, health staff identified benefits to themselves and patients. Barriers to implementing IPs were discussed.


Phase behavior of 2,3-disubstituted methyl octadecanoate monolayers at the air-water interface.
The phase behavior of 2,3-disubstituted methyl octadecanoate monolayers at the air-water interface is studied by film balance and a Brewster angle microscope (BAM). The comparison of the surface pressure-molecular area (pi-A) isotherms with the corresponding BAM images provides information on the phase behavior of the monolayers. Variations in the phase behavior of different 2,3-disubstituted methyl octadecanoate monolayers can be correlated with the size of the headgroups, the interactions between the polar molecular moieties and the subphase, and the intermolecular interactions. The enlarging of the headgroups makes forming a condensed monolayer difficult for the molecules, even after introduction of substituents giving rise to the formation of hydrogen bonds between the molecules, which may balance the steric repulsion and stabilize the monolayers. Model calculations of the two-dimensional lattice structure of the 2,3-disubstituted methyl octadecanoates on basis of the pg and p1 space group are performed and correspond well with the experimental results.


The electronic house call. Consequences of telemedicine consultations for physicians, patients, and society.
Within the next 10 years, telemedicine technology is likely to become widely available in physicians' offices and patients' homes. Rather than describing the technology of telemedicine, we discuss the implications of its widespread use among physicians, patients, and society as a whole, focusing on that mainstay of communication between physician and patient, the "electronic house call." After exploring the new relationship that is likely to develop between patients and physicians as a consequence of the use of telemedicine, we discuss the broader economic, legal, and political consequences of the widespread use of this technology.


Spontaneous pneumomediastinum: Case presentation to a college student health clinic.
The author describes a case of spontaneous pneumomediastinum (SPM) in a 19-year-old man presenting to a college student health clinic. The author also provides a review on SPM, including clinical manifestations, diagnostic evaluation, and management.


Evidence for age-dependent changes in Sertoli cell androgen receptor concentration.
Cytosol and nuclear receptor concentrations in Sertoli cells isolated from the testes of 15-, 25-, and 35-day-old rats were measured using hydroxylapatite separation procedures. In these cells the mean Kd of the cytosol receptor for methyltrienolone (3H-R1881) ranged between 2.3 and 2.9 nM, and the concentration of cytosol androgen receptor per mg Sertoli cell DNA increased over the 15-to 35-day age interval. However, when the data were expressed per mg cytosol protein, no increase was observed. The increase in receptor concentration per mg DNA paralleled the increase in cytosol protein/DNA ratio. The concentration of androgen receptor per mg DNA in nuclear extracts also increased with age. Consequently, total Sertoli cell androgen receptor increases over the time interval in which meiosis is first completed in the testis.


Synthesis of silver sulfide modified carbon materials for adsorptive removal of dibenzothiophene in n-hexane.
Carbon nanotube (CNT) and graphene oxide (GO) as common nanostructures were modified with silver sulfide (Ag2S) using chemical vapor deposition. The raw and modified materials were tested for the removal of Dibenzothiophene (DBT) from a model fuel in batch mode adsorption experiments. The maximum adsorption capacities of DBT were 52.18 and 49.65 mg g-1, using CNT-Ag2S and GO-Ag2S, respectively. The adsorption isotherm was modeled using Freundlich, Langmuir and Temkin models using linear and non-linear regression. The squared correlation coefficient (R2) and HYBRID error function were used to determine the best adsorption model. IR spectroscopy was used to study the DBT adsorption mechanism, and it was found that the DBT molecules lie flat on the surface of the developed adsorbents. Significant improvement was achieved in the adsorption of DBT using CNT-Ag2S and GO-Ag2S, where the maximum adsorption capacity increased by 127% and 117% respectively, which indicates a stronger interaction between DBT and the modified adsorbents.


Current-limiting amplifier for high speed measurement of resistive switching data.
Resistive switching devices, important for emerging memory and neuromorphic applications, face significant challenges related to the control of delicate filamentary states in the oxide material. As a device switches, its rapid conductivity change is involved in a positive feedback process that would lead to runaway destruction of the cell without current, voltage, or energy limitation. Typically, cells are directly patterned on MOS transistors to limit the current, but this approach is very restrictive as the necessary integration limits the materials available as well as the fabrication cycle time. In this article, we propose an external circuit to cycle resistive memory cells, capturing the full transfer curves while driving the cells in a way that suppresses runaway transitions. Using this circuit, we demonstrate the acquisition of 105 I, V loops per second without using on-wafer current limiting transistors. This setup brings voltage sweeping measurements to a relevant timescale for applications and enables many new experimental possibilities for device evaluation in a statistical context.


[How do general practitioners limit their prescriptions? A qualitative study based on a focus group].
There is no consensus on prescription of medicines in many situations in general medicine. The aim of this study was to identify the strategies used by general practitioners to limit prescriptions in order to make their prescriptions more effective. A mixed sample of general practitioners in terms of age and types of practice were interviewed using the focus group method until a sufficient number of data were obtained. Fourteen women and ten men aged between 32 and 64 years were interviewed by means of three group interviews. Various strategies were identified: the practitioner's attitude (rapid identification of the patient's needs, listening and evaluation of symptoms, support by physical examination) and the use of resources (reference tools and news) during the consultation; the importance of the conclusion of the consultation (written advice or visit report, review of the previous prescription) and explanation (reasons for limitation, reassurance, arguments, proposal of a follow-up visit). Limitation of prescriptions also depends on the practitioner's own reasons (initial and continued training, motivation and personal objectives, part of a peer group) but equally on the health care system (institutional, specialist support). The study highlights numerous approach to facilitate limitation of prescriptions: training and informing practitioners and patients, consultation management, promote communication at the heart of the health care system and policy-makers. Training organizations and health authorities could facilitate these strategies for the benefit of patients.


Taurine protects against NMDA-induced retinal damage by reducing retinal oxidative stress.
This study aimed to evaluate effect of TAU on NMDA-induced changes in retinal redox status, retinal cell apoptosis and retinal morphology in Sprague-Dawley rats. Taurine was injected intravitreally as pre-, co- or post-treatment with NMDA and 7 days post-treatment retinae were processed for estimation of oxidative stress, retinal morphology using H&E staining and retinal cell apoptosis using TUNEL staining. Treatment with TAU, particularly pre-treatment, significantly increased retinal glutathione, superoxide dismutase and catalase levels compared to NMDA-treated rats; whereas, the levels of malondialdehyde reduced significantly. Reduction in retinal oxidative stress in TAU pre-treated group was associated with significantly greater fractional thickness of ganglion cell layer within inner retina and retinal cell density in inner retina. TUNEL staining showed significantly reduced apoptotic cell count in TAU pre-treated group compared to NMDA group. It could be concluded that TAU protects against NMDA-induced retinal injury in rats by reducing retinal oxidative stress.


Changes in Caco-2 cells transcriptome profiles upon exposure to gold nanoparticles.
Higher efficacy and safety of nano gold therapeutics require examination of cellular responses to gold nanoparticles (AuNPs). In this work we compared cellular uptake, cytotoxicity and RNA expression patterns induced in Caco-2 cells exposed to AuNP (5 and 30nm). Cellular internalization was dose and time-dependent for both AuNPs. The toxicity was observed by colony forming efficiency (CFE) and not by Trypan blue assay, and exclusively for 5nm AuNPs, starting at the concentration of 200μM (24 and 72h of exposure). The most pronounced changes in gene expression (Agilent microarrays) were detected at 72h (300μM) of exposure to AuNPs (5nm). The biological processes affected by smaller AuNPs were: RNA/zinc ion/transition metal ion binding (decreased), cadmium/copper ion binding and glutathione metabolism (increased). Some Nrf2 responsive genes (several metallothioneins, HMOX, G6PD, OSGIN1 and GPX2) were highly up regulated. Members of the selenoproteins were also differentially expressed. Our findings indicate that exposure to high concentration of AuNPs (5nm) induces metal exposure, oxidative stress signaling pathways, and might influence selenium homeostasis. Some of detected cellular responses might be explored as potential enhancers of anti-cancer properties of AuNPs based nanomedicines.


Fauna of Cerambycidae (Insecta: Coleoptera) in Komaba Campus of the University of Tokyo, a highly urbanised area in Japan.
Urban green spaces play an important role in maintaining urban biodiversity in the Tokyo Metropolis, Japan. Plant-dependent insect assemblages such as Cerambycidae, in particular, are likely influenced by the existence of green spaces in Tokyo's urbanised environments. This study is the first comprehensive inventory of the cerambycid fauna in the Komaba Campus of the University of Tokyo. A cerambycid assemblage composed of a total of 25 species was recorded within the Komaba Campus site and compared to cerambycid assemblages in nine other green spaces distributed throughout Tokyo. The results indicated that the species number in the campus was similar to that recoded in a similar-sized green space in coastal Tokyo. Fewer cerambycid species were, however, found at the campus site than in larger-sized green spaces within Tokyo. Moreover, species compositions in urbanised areas were markedly different from those in suburbanised parks, mountains and forests within Tokyo.


An assessment of inflammation in the reservoir after restorative proctocolectomy with ileoanal ileal reservoir.
The significance of inflammation of the mucosa of the ileal reservoir after restorative proctocolectomy is not known although in some cases it appears to be associated with symptoms when the condition has been referred to as pouchitis. This investigation has aimed to determined the prevalence of inflammation, to define pouchitis and to examine some factors which might be related to inflammation. Mucosal biopsies from the ileal reservoir were studied in 90 patients at up to 62 months after closure of the ileostomy. A histological grading system (0-6) was used to assess the severity of inflammation. Some degree of chronic and acute inflammation was found in 87% and 30% of cases respectively. The prevalence of a grade of 4 or more was 23% and 3.5%. There was a correlation between severity of chronic and acute inflammation. Severe histological acute inflammation (grade 4-6) was associated with sigmoidoscopic features of inflammation and with increased frequency of defaecation. Of 55 patients sigmoidoscoped by one clinician, 6 (11%) had pouchitis which was characterised by macroscopic inflammation of the reservoir, diarrhoea and a histological grade of 4 or more. The severity of chronic inflammation was not related to frequency of defaecation. Histological inflammation could not be correlated with the type of reservoir, residual volume after evacuation of a known volume of stool substitute introduced per anum into the reservoir or compliance of the reservoir. Acute inflammation was significantly more severe in patients with ulcerative colitis than in those with familial adenomatous polyposis.


Associations of ABO blood type and galactose-deficient immunoglobulin A1 with adverse outcomes in patients with IgA nephropathy.
Both ABO blood group antigens and pathogenic immunoglobulin A1 (IgA1) in patients with IgA nephropathy (IgAN) are influenced by modifications of N-acetylgalactosamine and galactose. The purpose of this study was to assess whether ABO blood type is associated with galactose-deficient IgA1 (Gd-IgA1) in the progression of kidney disease in patients with IgAN. We enrolled 1313 IgAN patients with a median of 44 months follow-up and measured the plasma Gd-IgA1 levels. Multivariate Cox regression models were used to estimate the association between all variables and adverse outcomes. Using the propensity score matching method, 718 IgAN patients with blood type either A or B were selected, and their data were used to assess the association of blood type and Gd-IgA1/serum complement 3 (sC3) with outcomes. We found that the risk of adverse outcomes was significantly higher in patients with blood type A than in those with type B (hazard ratio = 1.82, 95% confidence interval 1.23-2.71; P = 0.003) after multivariate adjustment. The Gd-IgA1 levels showed trends similar to the multivariate-adjusted event-free curves for the blood types. However, this higher risk of adverse outcomes in type A than in type B patients was no longer significant after the addition of Gd-IgA1/sC3 to the model. IgAN patients with blood type A had a higher risk of adverse outcomes than those with type B, and this risk was associated with Gd-IgA1/sC3. Thus, the ABO blood type may provide a reference for the prognostic factors for individuals with IgAN.


Thermoresponsive Bentonite for Water-Based Drilling Fluids.
As an important industrial material, bentonite has been widely applied in water-based drilling fluids to create mud cakes to protect boreholes. However, the common mud cake is porous, and it is difficult to reduce the filtration of a drilling fluid at high temperature. Therefore, this paper endowed bentonite with a thermo response via the insertion of N-isopropylacrylamide (NIPAM) monomers. The interaction between NIPAM monomers and bentonite was investigated via Fourier infrared spectroscopy (FTIR), isothermal adsorption, and X-ray diffraction (XRD) at various temperatures. The results demonstrate that chemical adsorption is involved in the adsorption process of NIPAM monomers on bentonite, and the adsorption of NIPAM monomers accords with the D-R model. With increasing temperature, more adsorption water was squeezed out of the composite when the temperature of the composite exceeded 70 °C. Based on the composite of NIPAM and bentonite, a mud cake was prepared using low-viscosity polyanionic cellulose (Lv-PAC) and initiator potassium peroxydisulfate (KPS). The change in the plugging of the mud cake was investigated via environmental scanning electron microscopy (ESEM), contact angle testing, filtration experiments, and linear expansion of the shale at various temperatures. In the plugging of the mud cake, a self-recovery behavior was observed with increasing temperature, and resistance was observed at 110 °C. The rheology of the drilling fluid was stable in the alterative temperature zone (70-110 °C). Based on the high resistance of the basic drilling fluid, a high-density drilling fluid (ρ = 2.0 g/cm3) was prepared with weighting materials with the objective of drilling high-temperature formations. By using a high-density drilling fluid, the hydration expansion of shale was reduced by half at 110 °C in comparison with common bentonite drilling fluid. In addition, the rheology of the high-density drilling fluid tended to be stable, and a self-recovery behavior was observed.


CTLA-4 and CD4+ CD25+ regulatory T cells inhibit protective immunity to filarial parasites in vivo.
The T cell coinhibitory receptor CTLA-4 has been implicated in the down-regulation of T cell function that is a quintessential feature of chronic human filarial infections. In a laboratory model of filariasis, Litomosoides sigmodontis infection of susceptible BALB/c mice, we have previously shown that susceptibility is linked both to a CD4+ CD25+ regulatory T (Treg) cell response, and to the development of hyporesponsive CD4+ T cells at the infection site, the pleural cavity. We now provide evidence that L. sigmodontis infection drives the proliferation and activation of CD4+ Foxp3+ Treg cells in vivo, demonstrated by increased uptake of BrdU and increased expression of CTLA-4, Foxp3, GITR, and CD25 compared with naive controls. The greatest increases in CTLA-4 expression were, however, seen in the CD4+ Foxp3- effector T cell population which contained 78% of all CD4+ CTLA-4+ cells in the pleural cavity. Depletion of CD25+ cells from the pleural CD4+ T cell population did not increase their Ag-specific proliferative response in vitro, suggesting that their hyporesponsive phenotype is not directly mediated by CD4+ CD25+ Treg cells. Once infection had established, killing of adult parasites could be enhanced by neutralization of CTLA-4 in vivo, but only if performed in combination with the depletion of CD25+ Treg cells. This work suggests that during filarial infection CTLA-4 coinhibition and CD4+ CD25+ Treg cells form complementary components of immune regulation that inhibit protective immunity in vivo.


PCR-restriction endonuclease analysis for identification and strain typing of Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium based on polymorphisms in IS1311.
Point mutations in the IS 1311 sequences from sheep and cattle strains of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) and M. avium subsp. avium (M. avium) were targeted to develop a polymerase chain reaction (PCR) that would be useful in the diagnosis and control of Johne's disease. Candidate PCR tests were evaluated for sensitivity, specificity and ease of interpretation of the restriction endonuclease analysis (REA) products. One IS 1311 PCR, amplifying a 608 base pair product, was shown to be suitable when the amplified product was digested with Hinf I and Mse I. The PCR detected 50 fg of template DNA from M. paratuberculosis strain 316 V, the equivalent of 10 organisms. The test was evaluated further using purified DNA from M. paratuberculosis and M. avium isolates and diagnostic samples including primary radiometric cultures. All 89 M. paratuberculosis samples were correctly identified and typed according to host species or IS 900 restriction fragment length polymorphism (RFLP) type. All 28 isolates of M. avium were also correctly identified. A second PCR/REA strategy based on a shorter fragment of IS 1311 was developed for formalin-fixed paraffin-embedded tissue samples. It correctly differentiated sheep and cattle strains of M. paratuberculosis in 27 tissue samples in which acid fast bacilli had been observed in Ziehl Neelsen stains and in which sufficient amplified product was present for REA with Hinf I. Both tests were specific for M. paratuberculosis when tested against 24 other mycobacterial species. These simple and rapid tests can be used on a range of diagnostic samples for the confirmation of Johne's disease and will be of benefit in control and eradication programmes for this disease.


Chemical shift encoded imaging of hyperpolarized (13) C pyruvate.
To demonstrate a reconstruction technique for separating signal from different hyperpolarized carbon-13 metabolites. A reconstruction method is described for chemical shift encoded separation of the signal from pyruvate and its downstream metabolites. This method uses consistency of the data with the signal model rather than an additional free-induction decay (FID) acquisition to estimate the B0 offset. Compressed sensing was also integrated into the reconstruction allowing reconstruction of metabolite images from undersampled datasets. The performance of the reconstruction was assessed using thermal phantoms, digital phantoms, and in vivo hyperpolarized [1-(13) C] pyruvate experiments. Thermal and digital phantoms indicate that metabolite separation is feasible given Signal-to-noise ratio > 5 and an initial B0 offset estimate within -105 Hz to 90 Hz of the actual B0 offset. In vivo comparisons to an existing FID calibrated reconstruction show improved fidelity in regions with significant field map inhomogeneity provided that these field map variations are accounted for using an additional proton acquisition. Prospectively and retrospectively undersampled studies show acceleration factors of 2 are feasible using compressed sensing. A reconstruction framework for the separation of signal from pyruvate and its downstream metabolites is shown. This reconstruction eliminates the need to acquire additional calibration FID acquisition and allows acceleration through compressed sensing.


Membrane composition influences the activity of in vitro refolded human vitamin K epoxide reductase.
Human vitamin K epoxide reductase (hVKOR) is an integral membrane protein responsible for the maintenance of reduced vitamin K pools, a prerequisite for the action of γ-glutamyl carboxylase and hence for hemostasis. Here we describe the recombinant expression of hVKOR as an insoluble fusion protein in Escherichia coli, followed by purification and chemical cleavage under denaturing conditions. In vitro renaturation and reconstitution of purified solubilized hVKOR in phospholipids could be established to yield active protein. Crucially, the renatured enzyme is inhibited by the powerful coumarin anticoagulant warfarin, and we demonstrate that enzyme activity depends on lipid composition. The completely synthetic system for protein production allows a rational investigation of the multiple variables in membrane protein folding and paves the way for the provision of pure, active membrane protein for structural studies.


Symmetric gangrene of the extremities in late renal failure: a case report and review of the literature.
Medical calcification of arteries is common in chronic renal failure. We report on a patient with extensive calcification of the arterial media who developed symmetrical acral gangrene and severe cardiac failure shortly after cadaveric renal transplantation. At necropsy, the medial calcification was found to be accompanied by extensive intimal proliferation and multiple antemortem fractures, some healing by callus formation. Such medial calcification, which is similar to Monckeberg's sclerosis, affected all systemic arteries except the aorta, pulmonary artery and transplanted renal artery. Six years before this terminal illness he had undergone total parathyroidectomy for osteitis fibrosa associated with ruptured tendons. We review previous reports of patients with the syndrome of acral gangrene in azotaemic renal failure and discuss the histopathological features and pathogenesis in relation to the unusual features of our patient.


Metabolomic and antioxidant analyses of Salvia miltiorrhiza Bunge and Salvia prattii Hemsl. seeds.
Salvia miltiorrhiza and Salvia prattii seeds are rich in metabolites that are beneficial to human health and can be utilised as nutritional supplements. In this study, UPLC-MS and GC-MS based on extensively focused metabolomics were used to compare the seed metabolomics of the two species. LC-MS detected 118 metabolites, primarily Lipids and phenylpropanoids. GC- MS detected a total of 188 metabolites, mainly organic acids and their derivatives, of which Salvia prattii seeds contain high levels of nutrients. In addition, we experimentally determined antioxidant activity of two Salvia species, and the results showed that the antioxidant activity of Salvia prattii seeds was about twice as high as that of Salvia miltiorrhiza seeds. We used WGCNA to group the metabolites, and found the central metabolites in the focal modules including flavonoids and terpenoids. Our study contributes valuable knowledge for future research on the chemical makeup of Salvia prattii seeds.


Global circular RNA expression profile of human gastric cancer and its clinical significance.
Circular RNAs (circRNAs) are a new class of noncoding RNAs. However, the expression profile and clinical significance of circRNAs in human gastric cancer is unclear. The global circRNA expression profile in human gastric cancer was measured by circRNA microarray. Hsa_circ_0014717, one of the most downregulated circRNAs in microarray, was selected as a targeted circRNA to explore its levels in gastric tissues and gastric juice. Freeze-thaw experiment and incubation experiment confirmed the stability of gastric juice circRNAs. A total of 308 circRNAs, including 107 (34.74%) upregulated and 201 (65.26%) downregulated circRNAs, were found significantly aberrantly expressed in gastric cancer tissues. The top ten upregulated in gastric cancer tissues were hsa_circ_0035445, hsa_circ_0003789, hsa_circ_0063809, hsa_circ_0074362, hsa_circ_0006282, hsa_circ_0011107, hsa_circ_0084606, hsa_circ_0005556, hsa_circ_0050547, and hsa_circ_0006470, while the top ten downregulated ones were hsa_circ_0007099, hsa_circ_0001897, hsa_circ_0007707, hsa_circ_0008832, hsa_circ_0001546, hsa_circ_0002089, hsa_circ_0004680, hsa_circ_0000154, hsa_circ_0004458, and hsa_circ_0008394. The hot-point chromosomes were chr1, chr2, chr3, chr9, and chr17. Hsa_circ_0014717 was significantly downregulated in 77.2% (74/96) gastric cancer tissues. Its levels in gastric cancer tissues were related to tumor stage (P = 0.037), distal metastasis (P = 0.048), tissue carcinoembryonic antigen (P = 0.001), and carbohydrate antigen 19-9 expression (P = 0.021). More importantly, hsa_circ_0014717 can stably exist in human gastric juice; and its nature meets the requirements of clinical detection. Our study uncovered the circRNA expression profile in human gastric cancer. Moreover, some circRNAs can stably exist in human body fluid, and has the potential to be used as novel biomarkers for the screening of high-risk gastric cancer patients.


Glucagon-like peptide-1 receptor agonists for type 2 diabetes: A rational drug development.
Today, glucagon-like peptide-1 (GLP-1) receptor agonists are established glucose-lowering drugs used in the management of type 2 diabetes. Their development emerged from the understanding that a combined islet dysfunction comprising of impaired insulin secretion and exaggerated glucagon secretion is the key defect of hyperglycemia. GLP-1 was shown to target these defects, and after the discovery that dipeptidyl peptidase-4 inactivates native GLP-1, several different dipeptidyl peptidase-4-resistant GLP-1 receptor agonists have been developed. They are administered subcutaneously, but show differences in molecular structure, molecular size and pharmacokinetics, the latter allowing twice-daily, once-daily or once-weekly administration. They have been shown to be efficient in reducing both glycated hemoglobin and bodyweight, and to be safe and highly tolerable. Cardiovascular outcomes trials have shown them to be neutral or beneficial. GLP-1 receptor agonists are positioned as add-ons to metformin alone or in combination with oral agents in the clinical paradigm. They are also efficient when combined with insulin, and fixed dose combinations with long-acting insulin have been developed. Recent development includes a very long administration schedule and oral availability. The research from the first demonstration of the antidiabetic action of GLP-1 in the early 1990s to the enormously accumulated data today represents a successful and rational development, which has been characterized by focused perseverance to establish this therapy in the management of type 2 diabetes.


Common gamma chain cytokines: dissidence in the details.
Cytokines of the common cytokine-receptor gamma-chain (gamma(c)) family are essential for the development and maintenance of lymphocytes. Herein, we will focus on the roles of interleukin-2 (IL-2), IL-7, IL-15 and IL-21, in the orchestration of CD8 T cell responses. Among these cytokines, IL-7 has emerged as a master regulator of survival of immature and mature T lymphocytes, while IL-2, IL-15 and IL-21 appear to have specific functions in T cell homeostasis and differentiation. Hence, the gamma(c) has evolved as an elegant anchor through which related cytokines regulate distinct biological responses in T cells.


Semiordered Hierarchical Metallic Network for Fast and Large Charge-Induced Strain.
Nanoporous metallic actuators for artificial muscle applications are distinguished by combining the low operating voltage, which is otherwise reserved for polymer-based actuators with interesting values of strain amplitude, strength, and stiffness that are comparable of those of piezoceramics. We report a nanoporous metal actuator with enhanced strain amplitude and accelerated switching. Our 3D macroscopic metallic muscle has semiordered and hierarchical nanoporous structure, in which μm-sized tubes align perpendicular with the sample surface, while nm-sized ligaments consist of the tube walls. This nanoarchitecture combines channels for fast ion transportation with large surface area for charge storage and strain generation. The result is a record reversible strain amplitude of 1.59% with a strain rate of 8.83 × 10-6 s-1 in the field of metallic based actuators. A passive hydroxide layer is self-grown on the metal surface, which not only contributes a supercapacitive layer, but also stabilizes the nanoporous structure against coarsening, which guarantees sustainable actuation beyond ten-thousand cycles.


Current Trends and Future Directions for Outpatient Total Joint Arthroplasty: A Review of the Anesthesia Choices and Analgesic Options.
The aging population and the obesity epidemic have led to an increased rate of joint arthroplasty procedures, specifically total knee arthroplasty and total hip arthroplasty. These surgeries are associated with increased hospital length of stay and, consequently, higher costs. Despite the benefits of outpatient surgery, only a small percentage of total joint arthroplasties (TJAs) are done in this manner. We reviewed the most up-to-date trends for outpatient TJA and discussed essential factors for a successful outpatient program, including the proper patient selection process and best available anesthetic and analgesic options, along with their risks and benefits. Risk stratification tools, such as the Outpatient Arthroplasty Risk Assessment, are helpful for predicting outcomes regarding outpatient TJA, and neuraxial anesthesia should be considered to minimize complications and facilitate early discharge. A multimodal analgesia regimen could be effective for pain management in outpatient TJA, and the currently recommended peripheral nerve blocks for total hip arthroplasty and total knee arthroplasty are the fascia iliaca compartment block and adductor canal block, respectively. However, blocks should be carefully considered for outpatient procedures. Enhanced recovery after surgery (ERAS) protocols help to guide perioperative care teams and allow for improved patient recovery, decreased length of stay, and increased patient satisfaction.


Stretchable and flexible high-strain sensors made using carbon nanotubes and graphite films on natural rubber.
Conventional metallic strain sensors are flexible, but they can sustain maximum strains of only ~5%, so there is a need for sensors that can bear high strains for multifunctional applications. In this study, we report stretchable and flexible high-strain sensors that consist of entangled and randomly distributed multiwall carbon nanotubes or graphite flakes on a natural rubber substrate. Carbon nanotubes/graphite flakes were sandwiched in natural rubber to produce these high-strain sensors. Using field emission scanning electron microscopy, the morphology of the films for both the carbon nanotube and graphite sensors were assessed under different strain conditions (0% and 400% strain). As the strain was increased, the films fractured, resulting in an increase in the electrical resistance of the sensor; this change was reversible. Strains of up to 246% (graphite sensor) and 620% (carbon nanotube sensor) were measured; these values are respectively ~50 and ~120 times greater than those of conventional metallic strain sensors.


Noninvasive measurement of cardiac output in hemodialysis patients by task force monitor: a comparison with the Transonic System.
Cardiovascular disease is the leading cause of morbidity and mortality in maintenance hemodialysis (MHD) patients. The Transonic (TRS; Transonic Systems, Ithaca, NY) device is frequently used for determination of cardiac output (CO) by an indicator dilution technique. The Task Force Monitor (TFM; CN Systems, Graz, Austria) has gained attention as noninvasive tool for continuous beat-to-beat assessment of cardiovascular variables, including CO by impedance cardiography. Despite its use in cardiology and intensive care settings, the TFM has yet not been validated in dialysis patients. This study compares CO measurements in 12 MHD patients by TFM and TRS. Bland-Altman and regression analysis were used. CO was measured simultaneously by TRS and TFM. Average CO was 5.4 L/min by TRS and 5.0 L/min by TFM, respectively. Bland-Altman analysis revealed no significant systematic differences between the two methods (mean difference: 0.4 L/min; SD: 0.6; p > 0.05). Linear regression analysis showed significant correlation between both techniques (r = 0.802, p = 0.002). The SD of mean individual CO values was 1.1 L/min with TRS and 0.8 L/min with TFM, respectively.CO measured by TFM and TRS does not differ significantly, thus making the TFM an attractive noninvasive tool for the continuous beat-to-beat assessment of CO in MHD patients.


Occurrence of the siphonophore Muggiaea atlantica in Scottish coastal waters: source or sink?
We applied the concept of source-sink dynamics to investigate a recent (1999-2013) increase in the occurrence of the siphonophore Muggiaea atlantica in Scottish coastal waters. Our aim was to determine whether this change represented the establishment of resident populations (i.e. "sources"), or transient populations reliant on immigration (i.e. "sinks"). First, we show that local production was not always sufficient to account for recruitment (a "source" prerequisite), suggesting reliance on immigration (a "sink" prerequisite). Using variation partitioning, we then discriminated between the exclusive effects of immigration [indexed by the European Slope Current (ESC)] and local production (indexed by local sea temperature and food availability). On the west coast (Loch Ewe), interannual variability in the species' abundance was determined by, in order of increasing importance: (i) suitable local environmental conditions (13%); (ii) the role of the ESC in modulating these conditions (20%); and (iii) immigration via the ESC (29%). These results provided a strong indication that Loch Ewe represents a sink habitat for M. atlantica. However, on the east coast (Stonehaven) our results were less conclusive, probably due to the less direct influence of the ESC. For both locations, we suggest that low winter temperatures prevented overwintering, necessitating annual re-colonization via immigration.


Plasma and aqueous humur ascorbic acid levels in people with cataract from diverse geographical regions of Nepal.
Ascorbic acid in aqueous humor of the eye has a role as an antioxidant in delaying cataract formation. In a cross-sectional comparative study, ascorbic acid levels from aqueous humor and venous blood obtained from patients with cataracts in mountainous regions (1,300-2,000 meters) and subtropical lowlands (<100 meters) of Nepal were measured spectrophotometrically with the objective of comparing the levels of aqueous humor ascorbic acid in patients with cataracts from these two diverse geographical regions. Of 131 patients included in the study, 59 were from the mountainous region of Nepal (Group A) and 72 were from the subtropical lowlands (Group B). The mean (+/- SD) plasma ascorbic acid level (mg%) of the subjects in Group A was 0.65 (+/- 0.20) and Group B was 0.85 (+/- 0.31). The aqueous ascorbic acid levels ranged from 12-28 mg % (mean +/-SD = 17.5+/-4.52) and 14-50 mg % (mean +/-SD =23.47+/-8.66) in Groups A and B, respectively. The difference of the plasma and aqueous humor ascorbic acid levels between the two groups was statistically significant (p<0.01). The plasma and aqueous humor ascorbic acid levels in the people with cataracts from the mountainous region of Nepal were significantly lower than those from subtropical lowlands. This may be because of the effect of high altitude and ultraviolet rays on ascorbic acid metabolism.


Similar Fecal Microbiota Signatures in Patients With Diarrhea-Predominant Irritable Bowel Syndrome and Patients With Depression.
Patients with irritable bowel syndrome (IBS) often have psychiatric comorbidities. Alterations in the intestinal microbiota have been associated with IBS and depression, but it is not clear if there is a microbial relationship between these disorders. We studied the profiles of fecal microbiota samples from patients with IBS, depression, or comorbidities of IBS and depression; we determined the relationships among these profiles and clinical and pathophysiological features of these disorders. We used 454 pyrosequencing to analyze fecal microbiota samples from 100 subjects (40 with diarrhea-predominant IBS [IBS-D], 15 with depression, 25 with comorbidities of IBS and depression, and 20 healthy individuals [controls]), recruited at Peking University. Abdominal and psychological symptoms were evaluated with validated questionnaires. Visceral sensitivity was evaluated using a barostat. Colonic mucosal inflammation was assayed by immunohistochemical analyses of sigmoid tissue biopsy specimens. Fecal microbiota signatures were similar between patients with IBS-D and depression in that they were less diverse than samples from controls and had similar abundances of alterations. They were characterized by high proportions of Bacteroides (type I), Prevotella (type II), or nondominant microbiota (type III). Most patients with IBS-D or depression had type I or type II profiles (IBS-D had 85% type I and type II profiles, depression had 80% type I and type II profiles). Colon tissues from patients with type I or type II profiles had higher levels of inflammatory markers than colon tissues from patients with type III profiles. The level of colon inflammation correlated with the severity of IBS symptoms. Patients with IBS-D and depression have similar alterations in fecal microbiota; these might be related to the pathogenesis of these disorders. We identified 3 microbial profiles in patients that could indicate different subtypes of IBS and depression or be used as diagnostic biomarkers.


Substrate selective amide coupling driven by encapsulation of a coupling agent within a self-assembled hexameric capsule.
Encapsulation of a cationic carbodiimide condensing agent within a self-assembled hexameric capsule made of resorcin[4]arene units provides a nano-environment that efficiently steers the substrate selectivity in the amide synthesis reaction between carboxylic acids and primary amines. While in solution pairs of acids react similarly with a given amine, in the presence of the capsule the formation of the shorter amide is greatly favored.


Guideline appraisal with AGREE II: Systematic review of the current evidence on how users handle the 2 overall assessments.
The Appraisal of Guidelines for Research & Evaluation (AGREE) II instrument is the most commonly used guideline appraisal tool. It includes 23 appraisal criteria (items) organized within 6 domains and 2 overall assessments (1. overall guideline quality; 2. recommendation for use). The aim of this systematic review was twofold. Firstly, to investigate how often AGREE II users conduct the 2 overall assessments. Secondly, to investigate the influence of the 6 domain scores on each of the 2 overall assessments. A systematic bibliographic search was conducted for publications reporting guideline appraisals with AGREE II. The impact of the 6 domain scores on the overall assessment of guideline quality was examined using a multiple linear regression model. Their impact on the recommendation for use (possible answers: "yes", "yes, with modifications", "no") was examined using a multinomial regression model. 118 relevant publications including 1453 guidelines were identified. 77.1% of the publications reported results for at least one overall assessment, but only 32.2% reported results for both overall assessments. The results of the regression analyses showed a statistically significant influence of all domains on overall guideline quality, with Domain 3 (rigour of development) having the strongest influence. For the recommendation for use, the results showed a significant influence of Domains 3 to 5 ("yes" vs. "no") and Domains 3 and 5 ("yes, with modifications" vs. "no"). The 2 overall assessments of AGREE II are underreported by guideline assessors. Domains 3 and 5 have the strongest influence on the results of the 2 overall assessments, while the other domains have a varying influence. Within a normative approach, our findings could be used as guidance for weighting individual domains in AGREE II to make the overall assessments more objective. Alternatively, a stronger content analysis of the individual domains could clarify their importance in terms of guideline quality. Moreover, AGREE II should require users to transparently present how they conducted the assessments.


Controlled release of methotrexate from functionalized silica-gelatin aerogel microparticles applied against tumor cell growth.
Methotrexate functionalized silica-gelatin hybrid aerogel (SGM) was synthesized by the sol-gel method and co-gelation. The drug methotrexate (MTX) is covalently linked to the collagen molecules of the hybrid aerogel backbone by amide-bond. The characteristic MTX content of the functionalized hybrid aerogel is ca. 6 wt% by the dry weight. The micronization of SGM aerogel in water yields cell sized (d = 10-20 µm) particles. The cytotoxicity of these microparticles against tumor cell lines (SCC VII and HL-60) is unprecedentedly high, it is approximately equivalent to that of an equal dose of free (dissolved) MTX, as proved by in vitro experiments. Thus, the activity of MTX is intact after aerogel functionalization, and the mass specific cytotoxicity of SGM is high enough for medical applications. Drug release studies verified that MTX cannot be liberated from this drug delivery system solely by chemical hydrolysis, however, collagenase enzymatic activity releases MTX from the functionalized hybrid aerogel. The cytotoxicity of SGM towards various cancerous and non-cancerous cell lines correlates with the collagenase activities of cells. Therefore, conjugation with the hybrid aerogel provides a controlled release system for the antineoplastic agent MTX. The morphology of the delivery vehicle was chosen to adapt the size of cancer cells; thus the metastatic pathways of the tumor cells can get flooded.


Personality differences in hindsight bias.
Ten personality correlates of hindsight bias were tested in a study with 75 participants answering almanac-type knowledge questions. Participants showed hindsight bias when hindsight estimates were compared to foresight estimates (memory condition), when hindsight estimates were compared to foresight estimates of other participants (BS = between-subjects hypothetical condition), and when hindsight estimates were compared to foresight estimates in response to equally difficult control items (WS = within-subject hypothetical condition). The magnitude of hindsight bias in both hypothetical conditions was positively associated with the participant's field dependence and his or her tendency for favourable self-presentation (as measured by social desirability and impression management). Between-subjects hypothetical hindsight was associated with the participant's conscientiousness and need for predictability and control (as measured by a rigidity scale). In a multiple regression analysis, 39% of the variance in BS hypothetical hindsight, 24% of the variance in WS hypothetical hindsight, but no significant proportion of the variance in memory hindsight could be accounted for by personality measures. It is concluded that individual differences in hindsight bias exist and must be taken into account in a complete model of the effect.


Factors Affecting Subjective Symptoms in Children with Pes Planovalgus Deformity: A Study Using the Oxford Ankle Foot Questionnaire.
Idiopathic pes planovalgus is one of the most common foot deformities in children and adolescents. However, there is a discrepancy between subjective symptoms and radiographic severity in idiopathic planovalgus deformity, and very few studies have investigated this aspect. Further, the assessment of subjective symptoms in patients with pes planovalgus requires a quantitative scoring system for making meaningful comparisons, such as the Oxford Ankle Foot Questionnaire for Children (OxAFQ-c) and that for parents (OxAFQ-p). Therefore, the purpose of this study was to evaluate the factors affecting the symptoms of idiopathic planovalgus using the OxAFQ. All patients who were ≤18 years of age, had visited our clinic for the evaluation of pes planovalgus deformity, and had completed the OxAFQ were included in this study. The anteroposterior talo-first metatarsal, lateral talo-first metatarsal, and hallux valgus angles were measured on weight-bearing radiographs. The data were analyzed using a multiple regression model, with age, sex, and radiographic indices as explanatory variables. Overall, 123 patients were enrolled in this study, and 246 standing foot radiographs were evaluated along with scores in each domain of the OxAFQ. The factors affecting physical domain scores in the OxAFQ-c were female sex (p = 0.047) and the anteroposterior talo-first metatarsal angle (p = 0.033). Age of ≥10 years was a significant factor (p < 0.05) affecting all domains in both the OxAFQ-c and OxAFQ-p other than the physical domain score. Although pes planovalgus deformity is 3-dimensional, the forefoot abduction component of the deformity should be carefully assessed as it is closely related to subjective symptoms. Further, in female patients with a more severe anteroposterior talo-first metatarsal angle, an aggravation of symptoms with aging should be expected when managing pes planovalgus deformity. Prognostic Level IV. See Instructions for Authors for a complete description of levels of evidence.


The phylogeography of Adelie penguin faecal flora.
The gut of animals changes quickly from a totally sterile environment before birth to a numerous and highly diverse microbial community shortly after birth. However, few studies have examined the source of the bacteria colonizing the gut. We used a genetic based approach to investigate the distribution and acquisition of faecal microbial communities by Adelie penguins, Pygoscelis adeliae, breeding in the Ross Sea region of Antarctica by cloning a portion of the 16S rRNA gene and by automated ribosomal intergenic spacer analysis (ARISA). We hypothesized that bacteria were either acquired from the penguins' neighbours or inherited from their ancestors. Samples were collected from Adelie penguin breeding colonies at the north-western edge of the Ross Sea coast through to the southernmost Adelie penguin colonies on Ross Island. Most of the bacterial sequences we obtained were only distantly homologous with previously published sequences. Bacterial taxa appear to have a restricted distribution as the majority of 16S rRNA clones were isolated from only one or two hosts. Faecal bacterial community similarity was strongly negatively correlated with the genetic distance between hosts suggesting that bacterial communities are inherited. There was little support for a correlation between distance between collection sites and community similarity.


Photon irradiation using a water bath technique for treatment of confluent carcinoma in situ of the hand, digits, and nail bed: a case report.
Confluent squamous cell carcinoma in situ, or Bowen's disease, involving the hand, digit, and nail bed is rare and represents a significant therapeutic challenge. Surgical excision is recommended as first-line treatment but in cases of extensive disease can lead to unacceptable functional morbidity or cosmetic outcomes. Radiation therapy has been shown to be equally efficacious to surgery in the treatment of carcinoma in situ but its use has historically been limited due to concerns regarding toxicity. In this case report we present a novel therapeutic technique that may enable radiotherapy to be employed as a definitive treatment option in these challenging cases. A 75-year-old white man with a previous history of carcinoma in situ of his right hand previously treated with 5-fluorouracil presented with recurrent biopsy-proven confluent squamous cell carcinoma in situ of multiple surfaces of his right hand and digits with involvement of nail beds. To avoid extensive resection and possible amputation he was offered definitive external beam radiation therapy utilizing a water bath as a tissue-equivalent bolus material. This protocol enabled improved dose homogeneity to the target volume while minimizing acute toxicity. He experienced complete clinical resolution of the disease with only minimal acute edema and hyperpigmentation. Twenty months following treatment completion he remains disease-free with normal function and excellent cosmesis. Therapeutic radiation utilizing water as a tissue-equivalent bolus in this complicated case enabled definitive treatment of disease without compromising functional or cosmetic outcomes. Radiotherapy may therefore be an alternative and under-utilized approach to surgical excision in difficult-to-treat cases of carcinoma in situ.


High-speed AFM images of thermal motion provide stiffness map of interfacial membrane protein moieties.
The flexibilities of extracellular loops determine ligand binding and activation of membrane receptors. Arising from fluctuations in inter- and intraproteinaceous interactions, flexibility manifests in thermal motion. Here we demonstrate that quantitative flexibility values can be extracted from directly imaging the thermal motion of membrane protein moieties using high-speed atomic force microscopy (HS-AFM). Stiffness maps of the main periplasmic loops of single reconstituted water channels (AqpZ, GlpF) revealed the spatial and temporal organization of loop-stabilizing intraproteinaceous H-bonds and salt bridges.


BP1 is a negative modulator of definitive erythropoiesis.
Beta protein 1 (BP1), a human homeotic transcription factor, is expressed during hematopoeisis in the erythroid lineage. To determine the in vivo role of BP1 in erythropoiesis, we have undertaken two complementary approaches using enforced BP1 expression in both transgenic mice and embryonic stem (ES) cells. Despite repeated attempts, only one adult transgenic BP1 founder mouse among 121 mice was obtained. This mouse presumably survived due to transgene mosaicism because the transgene could not be transmitted. This mouse expressed BP1 and displayed splenomegaly, extramedullary erythropoiesis and severe amyloidosis A in the kidney, a phenotype compatible with thalassemia. Consistently, the presence of BP1 transgene in fetuses was associated with paleness and lethality. In ES cells, BP1 expression in primary differentiation appeared to antagonize adult beta-globin expression. In secondary differentiation, BP1 expression reduced significantly beta-globin gene expression in both primitive and definitive erythroid cells, whereas it impaired only the definitive erythroid cell differentiation. These studies showed that BP1 can negatively modulate adult beta-globin gene expression and definitive erythroid cell differentiation, and suggest that BP1 could play a role in thalassemia.


Detour test performance of cloned minipigs from three different clone populations.
Genetics, the uterine environment, maternal behavior, and rearing conditions can all influence animal behavioral phenotypes. Some studies on cloned pigs have found no differences between the behavioral patterns of cloned and non-cloned animals. Other studies conducted on dogs have reported similarities in the behavior of cloned subjects. This study evaluated the performance of 12 cloned minipigs from three different clone populations (A, B, C) in a detour test around symmetric and asymmetric barriers. We measured the detour time and patterns, in order to investigate the pigs' cognitive abilities.The detour time and the detour entry/exit pattern were recorded. All the animals tended to keep a fixed entry/exit pattern instead of modifying it to accommodate changes in the working set. Significant differences in detour time were found among the populations, with animals belonging to population B being faster than the others, and also within each population.Our study is one of the few to assess the cognitive abilities of cloned minipigs. The results indicate that even animals belonging to the same cloned population may develop different cognitive, hence behavioral characteristics. Whether cloning can be utilized to obtain similar behavioral phenotypes therefore remains a matter of debate.


[Modern diagnosis of Chlamydia trachomatis infections].
Chlamydia trachomatis is the most frequent sexually transmitted bacterial infection. Tests to diagnose C. trachomatis infections include growth on cell culture and assays detecting bacterial antigens or nucleic acids. Until recently, culture was considered the gold standard of testing because of its high specificity. However, the sensitivity of culture is limited. Nucleic acid amplification tests (NATs) have the highest sensitivity and are almost as specific as culture. They are best suited for screening. Chlamydial antigen EIAs are considerably less sensitive and specific, resulting in low positive predictive values, especially when low-prevalence populations are studied. Thus, positive results of chlamydia antigen EIAs should be confirmed by another test.


Expression of Plant Receptor Kinases in Tobacco BY-2 Cells.
Although more than 600 single-transmembrane receptor kinase genes have been found in the Arabidopsis genome, only a few of them have known physiological functions, and even fewer plant receptor kinases have known specific ligands. Ligand-binding analysis must be operated using the functionally expressed receptor form. However, the relative abundance of native receptor kinase molecules in the plasma membrane is often quite low. Here, we present a method for stable and functional expression of plant receptor kinases in tobacco BY-2 cells that allows preparation of microsomal fractions containing the receptor. This procedure provides a sufficient amount of receptor proteins while maintaining its ligand-binding activities.


Endemic bacterial meningitis in Sudanese children: aetiology, clinical findings, treatment and short-term outcome.
During the period April 1985 to November 1986 (18 months), 196 children (of age greater than 1 month) admitted to the Children's Emergency Hospital in Khartoum, Sudan, with clinical suspicion of meningitis/meningoencephalitis were followed up prospectively. Bacterial meningitis was diagnosed by culture, direct microscopy and/or antigen-detecting assays (co-agglutination and enzyme immunoassay) in 44 infants (25 Haemophilus influenzae type b, 8 Neisseria meningitidis, 7 Streptococcus pneumoniae, 3 enterobacteria and one mixed infection), aseptic meningitis in 52, cerebral malaria in 4 and febrile convulsions in 96. The majority of cases of bacterial meningitis were boys and 57% of those in whom H. influenzae was the commonest isolate were less than 1 year old. The presenting signs and symptoms are described as well as the transient and permanent short-term sequelae. The total mortality from bacterial meningitis was 19%, permanent neurological sequelae were seen in 26% of survivors. Prospective follow-up, including audiometry, of 35 children 1-2 months after discharge showed that 11% had hemiplegia and 20% had hearing impairment. The potential impact of vaccination against invasive H. influenzae infections is discussed.


Itraconazole versus fluconazole for prevention of fungal infections in patients receiving allogeneic stem cell transplants.
Prophylactic fluconazole prevents candidiasis; however, this drug has no activity against molds. We performed a randomized trial to determine whether prophylactic itraconazole prevents invasive mold infections (IMIs). A total of 304 patients receiving allogeneic stem cell transplants (SCT) were randomized to receive fluconazole (400 mg/d) or itraconazole (oral solution 2.5 mg/kg 3 times daily, or intravenous 200 mg daily) for 180 days after SC transplantation, or until 4 weeks after discontinuation of graft-versus-host disease (GVHD) therapy. Proven or probable invasive fungal infections (IFI) were evaluated by intent-to-treat and "on-treatment" analyses. More patients in the itraconazole arm developed hepatotoxicities, and more patients were discontinued from itraconazole because of toxicities or gastrointestinal (GI) intolerance (36% versus 16%, P <.001). Intent-to-treat analysis demonstrated no difference in the incidence of IFI during the intended study period (fluconazole 16% versus itraconazole 13%, P =.46); however, fewer patients in the itraconazole arm developed IFI on treatment (fluconazole 15% versus itraconazole 7%, P =.03). Itraconazole provided better protection against IMI (fluconazole 12% versus itraconazole 5%, P =.03), but similar protection against candidiasis (3% versus 2%, P =.69). There was no difference in overall or fungal-free survival. Itraconazole appears to prevent IMI in the subset of patients who tolerate the drug; however, toxicities and poor tolerability limit its success as prophylactic therapy.


Parameterizing dose-response models to estimate relative potency functions directly.
Many comparative analyses of toxicity assume that the potency of a test chemical relative to a reference chemical is constant, but employing such a restrictive assumption uncritically may generate misleading conclusions. Recent efforts to characterize non-constant relative potency rely on relative potency functions and estimate them secondarily after fitting dose-response models for the test and reference chemicals. We study an alternative approach of specifying a relative potency model a priori and estimating it directly using the dose-response data from both chemicals. We consider a power function in dose as a relative potency model and find that it keeps the two chemicals' dose-response functions within the same family of models for families typically used in toxicology. When differences in the response limits for the test and reference chemicals are attributable to the chemicals themselves, the older two-stage approach is the more convenient. When differences in response limits are attributable to other features of the experimental protocol or when response limits do not differ, the direct approach is straightforward to apply with nonlinear regression methods and simplifies calculation of simultaneous confidence bands. We illustrate the proposed approach using Hill models with dose-response data from U.S. National Toxicology Program bioassays. Though not universally applicable, this method of estimating relative potency functions directly can be profitably applied to a broad family of dose-response models commonly used in toxicology.


Medical applications with synchrotron radiation in Japan.
In Japan, various medical applications of synchrotron X-ray imaging, such as angiography, monochromatic X-ray computed tomography (CT), radiography and radiation therapy, are being developed. In particular, coronary arteriography (CAG) is quite an important clinical application of synchrotron radiation. Using a two-dimensional imaging method, the first human intravenous CAG was carried out at KEK in May 1996; however, further improvements of image quality are required in clinical practice. On the other hand, two-dimensional aortographic CAG revealed canine coronary arteries as clearly as those on selective CAG, and coronary arteries less than 0.2 mm in diameter. Among applications of synchrotron radiation to X-ray CT, phase-contrast X-ray CT and fluorescent X-ray CT are expected to be very interesting future applications of synchrotron radiation. For actual clinical applications of synchrotron radiation, a medical beamline and a laboratory are now being constructed at SPring-8 in Harima.


Enhancement of cytotoxicity of bleomycin by dithiocarbamates: formation of bis(dithiocarbamato) cu(II).
Properties of the reactions of dithiocarbamates and their Cu(II) or Fe(III) complexes with Ehrlich cells were determined and related to their effects on the inhibition of cell proliferation caused by bleomycin and Cu bleomycin. In complete culture medium containing Eagle's minimal essential medium plus Earles salts and 2.5% fetal calf serum, dimethyl- and diethyldithiocarbamates and their copper complexes inhibit cell proliferation and cause cell death. The copper complexes are more effective agents. Ferric tris-diethyldithiocarbamate is also a cytotoxic species. In contrast, when cells are exposed to dimethyldithiocarbamate or its copper complex in Ringer's buffer under metal-restricted condition, washed, and then placed in complete medium, the copper complex is much more active in inhibiting cell growth. The difference is magnified when dihydroxyethyldithiocarbamate and N-methylglucamine dithiocarbamate and their copper complexes are compared in complete media. Incubation of bleomycin or copper bleomycin with Ehrlich cells in Ringer's buffer with or without dimethyldithiocarbamate or bis-dimethyldithiocarbamato Cu(II) leads to no enhancement of cytotoxicity from combinations of agents, except when the two copper complexes are present. Diethyl- or dimethyldithiocarbamate readily extracts copper from Cu(II)bleomycin and iron from Fe(III)bleomycin when ethylacetate is present to remove the tris-dithiocarbamato Fe(III) complex from aqueous solution. When bis-dimethyldithiocarbamato Cu(II) is incubated with Ehrlich cells, copper is released from the complex and bound to high molecular weight and metallothionein fractions. A reductive mode of dissociation of the copper complexes in cells is supported by ESR experiments. Reactions of diethyl- and dimethyldithiocarbamato Cu(II) with thiol compounds demonstrates one possible mechanism of reduction of these complexes.


[Heavy metals in food--analytical control in Austria].
Cadmium, lead, mercury and arsenic were determined by means of atomic absorption spectrophotometry. The procedures used for digestion, extraction and concentration and the measuring technique are described. Nearly 1500 determinations of lead, 1200 determinations of mercury and arsenic, respectively, and 800 determinations of cadmium in foods permitted to calculate (taking into account the food habits) the dietary loads on the Austrian population. The loads per individual and month were: 5.9 mg lead (infants aged from 1--12 months: 0.18--2.5 mg), 2.02 mg cadmium (infants, 0.04--0.61 mg), 0.2 mg mercury and 0.9 mg arsenic. Whereas the cadmium intake was somewhat greater than the maximum value established by the FAO/WHO (1.6--2.0 mg/individual and month), the intakes of lead, mercury and arsenic were considerably lower than the maximum values.


Cyclophosphamide immunosuppression during lymphotropic herpesvirus infection in the guinea pig model. A histopathologic and virologic study.
Guinea pigs infected with lymphotropic herpesvirus (GPHLV) were given the immunosuppressive agent cyclophosphamide (Cy). All Cy-treated animals revealed the expected lymphoid depletion of spleen and lymph node B zones. Acute GPHLV infection of Cy-treated animals resulted in increased blood and spleen leukocyte viral infectivity titers and lymphoid tissue lesions containing cells positive for GPHLV antigen and intranuclear inclusions. During latent GPHLV infection, Cy treatment resulted in declining leukocyte viral infectivity titers without pathologic lesions. Morphologic data suggest that tissue histiocytic cells may be involved in the productive viral infection observed in Cy-immunosuppressed animals during acute GPHLV infection. During latency, however, infectious virus appears restricted to a Cy-sensitive, probably lymphoid, cell. This animal model appears useful for the study of lymphotropic viral infection during immunosuppression.


An improved high-throughput screening assay for tunicamycin sensitivity in Arabidopsis seedlings.
Tunicamycin (Tm) sensitivity assays are a useful method for studies of endoplasmic reticulum stress and the unfolded protein response in eukaryotic cells. While Tm sensitivity and Tm recovery assays have been previously described, these existing methods are time-consuming, labor intensive, and subjected to mechanical wounding. This study shows an improved method of testing Tm sensitivity in Arabidopsis using liquid Murashige and Skoog medium versus the traditional solid agar plates. Liquid medium bypasses the physical manipulation of seedlings, thereby eliminating the risk of potential mechanical damage and additional unwanted stress to seedlings. Seedlings were subjected to comparative treatments with various concentrations of Tm on both solid and liquid media and allowed to recover. Determination of fresh weight, chlorophyll contents analysis and qRT-PCR results confirm the efficacy of using liquid medium to perform quantitative Tm stress assays.


Raman difference spectroscopy: a non-invasive method for identification of oral squamous cell carcinoma.
The feasibility of shifted-excitation Raman difference spectroscopy (SERDS) as a label-free and non-invasive technique for an objective diagnosis of oral cancer (OSCC) was investigated by analyzing 12 ex vivo OSCC samples. 72 mean SERDS spectra from each three physiological tissue points and pathological lesions were correlated with the histo-pathological diagnosis. Principal component analysis (PCA) and linear discriminant analysis (LDA) showed excellent results with an area under the curve of 94.5% and a classification error of 9.7% (sensitivity: 86.1%; specificity: 94.4%). The SERDS Raman spectra of malignant and benignant tissues were discriminable with respect to the spectral features of proteins, lipids and nucleic acids. The presented method is capable of a highly accurate identification of OSCC. These findings suggest a high validity and reproducibility of SERDS combined with PCA and LDA analysis regarding oral cancer tissue.


Silica-Encapsulated Pt-Sn Intermetallic Nanoparticles: A Robust Catalytic Platform for Parahydrogen-Induced Polarization of Gases and Liquids.
Recently, a facile method for the synthesis of size-monodisperse Pt, Pt3 Sn, and PtSn intermetallic nanoparticles (iNPs) that are confined within a thermally robust mesoporous silica (mSiO2 ) shell was introduced. These nanomaterials offer improved selectivity, activity, and stability for large-scale catalytic applications. Here we present the first study of parahydrogen-induced polarization NMR on these Pt-Sn catalysts. A 3000-fold increase in the pairwise selectivity, relative to the monometallic Pt, was observed using the PtSn@mSiO2 catalyst. The results are explained by the elimination of the three-fold Pt sites on the Pt(111) surface. Furthermore, Pt-Sn iNPs are shown to be a robust catalytic platform for parahydrogen-induced polarization for in vivo magnetic resonance imaging.


Persistent paralysis in critically ill patients after the use of neuromuscular blocking agents.
Neuromuscular blocking agents (NMBAs), an important part of the pharmacologic armamentarium of the intensivist, have a long and admirable history of safety when used in the operating room for periods of time (almost always < 12 hrs). Since 1985, dozens of medical journals have reported a multitude of studies on persistent paralysis when these same agents are exported from the operating room to the ICU. Most of these reports are case presentations of patients who failed to move for days to weeks after discontinuation of NMBAs. These reports have led to concern about the appropriate use of NMBAs in the ICU. This article sorts through the issues surrounding persistent paralysis, and defines it as a short-term and a long-term problem. The short-term problem seems to have a pharmacologic explanation that is not difficult to correct. The long-term problem is much more complex and may have a toxic explanation that may also be more difficult to manage.


Testing the GPO waters.
When deciding whether to switch to a new group purchasing organization (GPO), consider the context of the opportunity presented, the experience and character of those associated with the new GPO, contract terms that can affect savings, and how savings will be measured.


Endocrine effects of duodenal-jejunal exclusion in obese patients with type 2 diabetes mellitus.
Duodenal-jejunal bypass liner (DJBL) is an endoscopically implantable device designed to noninvasively mimic the effects of gastrointestinal bypass operations by excluding the duodenum and proximal jejunum from the contact with ingested food. The aim of our study was to assess the influence of DJBL on anthropometric parameters, glucose regulation, metabolic and hormonal profile in obese patients with type 2 diabetes mellitus (T2DM) and to characterize both the magnitude and the possible mechanisms of its effect. Thirty obese patients with poorly controlled T2DM underwent the implantation of DJBL and were assessed before and 1, 6 and 10months after the implantation, and 3months after the removal of DJBL. The implantation decreased body weight, and improved lipid levels and glucose regulation along with reduced glycemic variability. Serum concentrations of fibroblast growth factor 19 (FGF19) and bile acids markedly increased together with a tendency to restoration of postprandial peak of GLP1. White blood cell count slightly increased and red blood cell count decreased throughout the DJBL implantation period along with decreased ferritin, iron and vitamin B12 concentrations. Blood count returned to baseline values 3months after DJBL removal. Decreased body weight and improved glucose control persisted with only slight deterioration 3months after DJBL removal while the effect on lipids was lost. We conclude that the implantation of DJBL induced a sustained reduction in body weight and improvement in regulation of lipid and glucose. The increase in FGF19 and bile acids levels could be at least partially responsible for these effects.


[Cerebral contrast medium extravasation after coronary angioplasty].
We report on an 82-year-old woman who suffered a sudden loss of consciousness, right-sided hemiparesis, and aphasia during a coronary angiographic procedure. Computed tomography (CT) of the brain performed immediately revealed an edema and extravascularly localized contrast media in the left hemisphere. Within 6 h, neurological symptoms had disappeared, and a second CT after 48 h revealed normal results. Usually, extravasation of contrast media affects the posterior circulation with cortical blindness. This case demonstrates that contrast media may affect the blood-brain barrier also outside the posterior circulation. If neurological symptoms occur during angiography, contrast media extravasation must be distinguished from embolism or vasospasm induced by the angiographic procedure.


High Precision FRET at Single-molecule Level for Biomolecule Structure Determination.
A protocol on how to perform high-precision interdye distance measurements using Förster resonance energy transfer (FRET) at the single-molecule level in multiparameter fluorescence detection (MFD) mode is presented here. MFD maximizes the usage of all "dimensions" of fluorescence to reduce photophysical and experimental artifacts and allows for the measurement of interdye distance with an accuracy up to ~1 Å in rigid biomolecules. This method was used to identify three conformational states of the ligand-binding domain of the N-methyl-D-aspartate (NMDA) receptor to explain the activation of the receptor upon ligand binding. When comparing the known crystallographic structures with experimental measurements, they agreed within less than 3 Å for more dynamic biomolecules. Gathering a set of distance restraints that covers the entire dimensionality of the biomolecules would make it possible to provide a structural model of dynamic biomolecules.


Brain interference: Revisiting the role of IFNγ in the central nervous system.
Interferon gamma (IFNγ) is a pro-inflammatory cytokine, first described as a secreted molecule capable of interfering with viral replication. Since then, numerous other important actions in the context of the immune response to invading pathogens (including those invading the brain) have been ascribed to this pleiotropic cytokine. Nevertheless, the precise role of IFNγ in neuropsychiatric and neurodegenerative disorders, and its possible contribution to the regulation of normal brain function, remains enigmatic. This review integrates and considers current knowledge about IFNγ actions with accumulating evidence of its importance on neurocytogenesis, synaptic plasticity and neurodegeneration within the framework of brain health and disease.