Simultaneous analysis of non-steroidal anti-inflammatory drugs using electrochemically controlled solid-phase microextraction based on nanostructure molecularly imprinted polypyrrole film coupled to ion mobility spectrometry.
A simple, rapid, and highly sensitive method for simultaneous analysis of anti-inflammatory drugs (naproxen, ibuprofen, and mefenamic acid) in diluted human serum was developed using the electrochemically controlled solid-phase microextraction coupled to ion mobility spectrometry. A conducting molecularly imprinted polymer film based on polypyrrole was synthesized for the selective uptake and release of drugs. The film was prepared by incorporation of a template molecule (naproxen) during the electropolymerization of pyrrole onto a platinum electrode using cyclic voltammetry method. The measured ion mobility spectrometry intensity was related to the concentration of analytes taken up into the films. The calibration graphs (naproxen, ibuprofen, and mefenamic acid) were linear in the range of 0.1-30 ng/mL and detection limits were 0.07-0.37 ng/mL and relative standard deviation was lower than 6%. On the basis of the results obtained in this work, the conducting molecularly imprinted polymer films as absorbent have been applied in the electrochemically controlled solid-phase microextraction and ion mobility spectrometry system for the selective clean-up and quantification of trace amounts of anti-inflammatory drugs in human serum samples. Scanning electron microscopy has confirmed the nano-structure morphology of the polypyrrole film.


Identification and characterisation of peptides from a boarfish (Capros aper) protein hydrolysate displaying in vitro dipeptidyl peptidase-IV (DPP-IV) inhibitory and insulinotropic activity.
Twenty-two novel dipeptidyl peptidase-IV (DPP-IV) inhibitory peptides (with IC50 values <200 µM) and fifteen novel insulinotropic peptides were identified in a boarfish protein hydrolysate generated at semi-pilot scale using Alcalase 2.4L and Flavourzyme 500L. This was achieved by bioassay-driven semi-preparative reverse phase-high performance liquid chromatography fractionation, liquid chromatography-mass spectrometry and confirmatory studies with synthetic peptides. The most potent DPP-IV inhibitory peptide (IPVDM) had a DPP-IV half maximal inhibitory concentration (IC50) value of 21.72 ± 1.08 µM in a conventional in vitro and 44.26 ± 0.65 µM in an in situ cell-based (Caco-2) DPP-IV inhibition assay. Furthermore, this peptide stimulated potent insulin secretory activity (1.6-fold increase compared to control) from pancreatic BRIN-BD11 cells grown in culture. The tripeptide IPV exhibited potent DPP-IV inhibitory activity (IC50: 5.61 ± 0.20 µM) comparable to that reported for the known DPP-IV inhibitor IPI (IC50: 3.20 µM). Boarfish proteins contain peptide sequences with potential to play a role in glycaemic management in vivo.


Bickerstaff's encephalitis and Miller Fisher syndrome associated with voltage-gated potassium channel and novel anti-neuronal antibodies.
GQ1b antibody (GQ1b-Ab) is detected in approximately two-thirds of sera of patients with Bickerstaffs encephalitis (BE). Whilst some of the remaining patients have antibodies to other gangliosides, many patients with BE are reported to be seronegative. Voltage-gated potassium channel antibody (VGKC-Ab) at high titer was detected during the diagnostic work-up of one patient with BE. Sera of an additional patient with BE and nine patients with Miller Fisher syndrome (MF) (all GQ1b-Ab positive) were investigated for VGKC-Ab and other anti-neuronal antibodies by radioimmunoprecipitation using 125I-dendrotoxin-VGKC and immunohistochemistry, respectively. Two patients with MF exhibited moderate titer VGKC-Abs. Regardless of positivity for VGKC or GQ1b antibodies, serum IgG of all patients with BE and MF reacted with the molecular layer and Purkinje cells of the cerebellum in a distinctive pattern. Voltage-gated potassium channel antibodies might be involved in some cases of BE or MF. The common staining pattern despite different antibody results suggests that there might be other, as yet unidentified, antibodies associated with BE and MF.


Biochemical typing of pathological prion protein in aging cattle with BSE.
The broad enforcement of active surveillance for bovine spongiform encephalopathy (BSE) in 2000 led to the discovery of previously unnoticed, atypical BSE phenotypes in aged cattle that differed from classical BSE (C-type) in biochemical properties of the pathological prion protein. Depending on the molecular mass and the degree of glycosylation of its proteinase K resistant core fragment (PrPres), mainly determined in samples derived from the medulla oblongata, these atypical cases are currently classified into low (L)-type or high (H)-type BSE. In the present study we address the question to what extent such atypical BSE cases are part of the BSE epidemic in Switzerland. To this end we analyzed the biochemical PrPres type by Western blot in a total of 33 BSE cases in cattle with a minimum age of eight years, targeting up to ten different brain regions. Our work confirmed H-type BSE in a zebu but classified all other cases as C-type BSE; indicating a very low incidence of H- and L-type BSE in Switzerland. It was documented for the first time that the biochemical PrPres type was consistent across different brain regions of aging animals with C-type and H-type BSE, i.e. independent of the neuroanatomical structure investigated. Taken together this study provides further characteristics of the BSE epidemic in Switzerland and generates new baseline data for the definition of C- and H-type BSE phenotypes, thereby underpinning the notion that they indeed represent distinct prion disease entities.


Determining thickness of films on a curved substrate by use of ellipsometric measurements.
A three-intensity measurement technique has been employed in polarizer-sample-analyzer imaging ellipsometry to measure the two-dimensional ellipsometric parameters of a coated cylindrical lens. Since azimuth deviation alpha of a polarizer can also be measured with this three-intensity measurement technique, we tilted a well-calibrated thin-film wafer to identify the orientation of the measured alphawith respect to the incident plane. Using the analytic property of this measurement technique, we can correct the deviation and determine the thickness profile of the thin film coated on a cylindrical lens.


A modeling exploration of how synaptic feedback to descending projection neurons shapes the activity of an oscillatory network.
Rhythmic activity which underlies motor output is often initiated and controlled by descending modulatory projection pathways onto central pattern generator (CPG) networks. In turn, these descending pathways receive synaptic feedback from their target CPG network, which can influence the CPG output. However, the mechanisms underlying such bi-directional synaptic interactions are mostly unexplored. We develop a reduced mathematical model, including both feed-forward and feedback circuitry, to examine how the synaptic interactions involving two projection neurons, MCN1 and CPN2, can produce and shape the activity of the gastric mill CPG in the crab stomatogastric nervous system. We use simplifying assumptions that are based on the behavior of the biological system to reduce this model down to 2 dimensions, which allows for phase plane analysis of the model output. The model shows a distinct activity for the gastric mill rhythm that is elicited when MCN1 and CPN2 are co-active compared to the rhythm elicited by MCN1 activity alone. Furthermore, the presence of feedback to the projection neuron CPN2 provides a distinct locus of pattern generation in the model which does not require reciprocally inhibitory interactions between the gastric mill CPG neurons, but is instead based on a half-center oscillator that occurs through a tri-synaptic pathway that includes CPN2. Our modeling results show that feedback to projection pathways may provide additional mechanisms for the generation of motor activity. These mechanisms can have distinct dependence on network parameters and may therefore provide additional flexibility for the rhythmic motor output.


Reminder Cards Improve Physician Documentation of Obesity But Not Obesity Counseling.
Physicians frequently fail to document obesity and obesity-related counseling. We sought to determine whether attaching a physical reminder card to patient encounter forms would increase electronic medical record (EMR) assessment of and documentation of obesity and dietary counseling. Reminder cards for obesity documentation were attached to encounter forms for patient encounters over a 2-week intervention period. For visits in the intervention period, the EMR was retrospectively reviewed for BMI, assessment of "obesity" or "morbid obesity" as an active problem, free-text dietary counseling within physician notes, and assessment of "dietary counseling" as an active problem. These data were compared to those collected through a retrospective chart review during a 2-week pre-intervention period. We also compared physician self-report of documentation via reminder cards with EMR documentation. We found significant improvement in the primary endpoint of assessment of "obesity" or "morbid obesity" as an active problem (42.5% versus 28%) compared to the pre-intervention period. There was no significant difference in the primary endpoints of free-text dietary counseling or assessment of "dietary counseling" as an active problem between the groups. Physician self-reporting of assessment of "obesity" or "morbid obesity" as an active problem (77.7% versus 42.5%), free-text dietary counseling on obesity (69.1% versus 35.4%) and assessment of "dietary counseling" as an active problem (54.3% versus 25.2%) were all significantly higher than those reflected in EMR documentation. This study demonstrates that physical reminder cards are a successful means of increasing obesity documentation rates among providers but do not necessarily increase rates of obesity-related counseling or documentation of counseling. Our study suggests that even with such interventions, physicians are likely under-documenting obesity and counseling compared to self-reported rates.


Function and biological roles of the Dickkopf family of Wnt modulators.
Dickkopf (Dkk) genes comprise an evolutionary conserved small gene family of four members (Dkk1-4) and a unique Dkk3-related gene, Dkkl1 (soggy). They encode secreted proteins that typically antagonize Wnt/beta-catenin signaling, by inhibiting the Wnt coreceptors Lrp5 and 6. Additionally, Dkks are high affinity ligands for the transmembrane proteins Kremen1 and 2, which also modulate Wnt signaling. Dkks play an important role in vertebrate development, where they locally inhibit Wnt regulated processes such as antero-posterior axial patterning, limb development, somitogenesis and eye formation. In the adult, Dkks are implicated in bone formation and bone disease, cancer and Alzheimer's disease.


A Mn-54 radiotracer study of Mn isotope solid-liquid exchange during reductive transformation of vernadite (δ-MnO2) by aqueous Mn(II).
We employed Mn-54 radiotracers to characterize the extent and dynamics of Mn atom exchange between aqueous Mn(II) and vernadite (δ-Mn(IV)O2) at pH 7.5 under anoxic conditions. Exchange of Mn atoms between the solid and liquid phase is rapid, reaching dynamic equilibrium in 2-4 days. We propose that during the initial stages of reaction, Mn atom exchange occurs through consecutive comproportionation-disproportionation reactions where interfacial electron transfer from adsorbed Mn(II) to lattice Mn(IV) generates labile Mn(III) cations that rapidly disproportionate to reform aqueous Mn(II) and solid-phase Mn(IV). Following nucleation of Mn(III)OOH phases, additional exchange likely occurs through electron transfer from aqueous Mn(II) to solid-phase Mn(III). Our results provide evidence for the fast and extensive production of transient Mn(III) species at the vernadite surface upon contact of this substrate with dissolved Mn(II). We further show that HEPES buffer is a reductant of lattice Mn(IV) in the vernadite structure in our experiments. The methods and results presented here introduce application of Mn-54 tracers as a facile tool to further investigate the formation kinetics of labile Mn(III) surface species and their impacts on Mn-oxide structure and reactivity over a range of environmentally relevant geochemical conditions.


Guinea pig kisspeptin neurons are depolarized by leptin via activation of TRPC channels.
Hypothalamic kisspeptin neurons are critical for driving reproductive function, but virtually nothing is known about their endogenous electrophysiological properties and the effects of leptin on their excitability. Therefore, we used the slice preparation from female guinea pigs to study the endogenous conductances and the effects of leptin on kisspeptin neurons. We targeted the arcuate kisspeptin neurons using visualized-patch whole-cell recording and identified kisspeptin neurons using immuocytochemical staining for kisspeptin or single cell RT-PCR. We also harvested dispersed arcuate neurons for analysis of expression of channel transcripts. Kisspeptin neurons exhibited a relatively negative resting membrane potential, and eighty percent of the neurons expressed a pacemaker current (h-current) and a T-type Ca(2+) current. Furthermore, the glutamate receptor agonist N-methyl D-aspartic acid depolarized and induced burst firing in kisspeptin neurons. Leptin activated an inward current that depolarized kisspeptin neurons and increased (burst) firing, but leptin hyperpolarized NPY neurons. Lanthanum, a TRPC-4,-5 channel activator, potentiated the leptin-induced inward current by 170%. The leptin-activated current reversed near -15 mV and was abrogated by the relatively selective TRPC channel blocker 2-APB. The leptin effects were also blocked by a Janus kinase inhibitor, a phosphatidylinositol 3 kinase inhibitor, and a phospholipase Cγ inhibitor. In addition, the majority of these neurons expressed TRPC1 and -5 and phospholipase Cγ1 based on single cell RT-PCR. Therefore, guinea pig kisspeptin neurons express endogenous pacemaker currents, and leptin excites these neurons via activation of TRPC channels. The leptin excitatory effects on kisspeptin neurons may be critical for governing the excitatory drive to GnRH neurons during different nutritional states.


Habitat compression and ecosystem shifts as potential links between marine heatwave and record whale entanglements.
Climate change and increased variability and intensity of climate events, in combination with recovering protected species populations and highly capitalized fisheries, are posing new challenges for fisheries management. We examine socio-ecological features of the unprecedented 2014-2016 northeast Pacific marine heatwave to understand the potential causes for record numbers of whale entanglements in the central California Current crab fishery. We observed habitat compression of coastal upwelling, changes in availability of forage species (krill and anchovy), and shoreward distribution shift of foraging whales. We propose that these ecosystem changes, combined with recovering whale populations, contributed to the exacerbation of entanglements throughout the marine heatwave. In 2016, domoic acid contamination prompted an unprecedented delay in the opening of California's Dungeness crab fishery that inadvertently intensified the spatial overlap between whales and crab fishery gear. We present a retroactive assessment of entanglements to demonstrate that cooperation of fishers, resource managers, and scientists could mitigate future entanglement risk by developing climate-ready fisheries approaches, while supporting thriving fishing communities.


Characterization of human NLZ1/ZNF703 identifies conserved domains essential for proper subcellular localization and transcriptional repression.
NET family members have recently emerged as important players in the development of multiple structures, from the trachea of fly larvae to the vertebrate eye and human breast cancers. However, their mechanisms of action are still poorly understood, and we lack a detailed characterization of their functional domains, as well as gene expression patterns-particularly in adult mammals. Here, we present a characterization of human NLZ1/ZNF703 (NocA-like zinc finger 1/Zinc finger 703), one of the two human NET family member genes. We show that the gene is ubiquitously expressed in adult human and mouse tissues, that three mRNA species with the same coding sequence are generated by alternative polyadenylation, and that the encoded protein contains six evolutionarily conserved domains, three of which are specific to NET proteins. Finally, we present functional evidence that these domains are necessary for proper subcellular distribution of and transcription repression by the NLZ1 protein, but not for its interaction with Groucho family co-repressors.


The effects of a high-energy diet on hippocampal function and blood-brain barrier integrity in the rat.
Cognitive impairment and Alzheimer's disease are linked with intake of a Western diet, characterized by high levels of saturated fats and simple carbohydrates. In rats, these dietary components have been shown to disrupt hippocampal-dependent learning and memory processes, particularly those involving spatial memory. Using a rat model, the present research assessed the degree to which consumption of a high-energy (HE) diet, similar to those found in modern Western cultures, produces a selective impairment in hippocampal function as opposed to a more global cognitive disruption. Learning and memory performance was examined following 90-day consumption of an HE-diet in three nonspatial discrimination learning problems that differed with respect to their dependence on the integrity of the hippocampus. The results showed that consumption of the HE-diet impaired performance in a hippocampal-dependent feature negative discrimination problem relative to chow-fed controls, whereas performance was spared on two discrimination problems that do not rely on the hippocampus. To explore the mechanism whereby consuming HE-diets impairs cognitive function, we investigated the effect of HE-diets on the integrity of the blood-brain barrier (BBB). We found that HE-diet consumption produced a decrease in mRNA expression of tight junction proteins, particularly Claudin-5 and -12, in the choroid plexus and the BBB. Consequently, an increased blood-to-brain permeability of sodium fluorescein was observed in the hippocampus, but not in the striatum and prefrontal cortex following HE-diet access. These results indicate that hippocampal function may be particularly vulnerable to disruption by HE-diets, and this disruption may be related to impaired BBB integrity.


Chondrotympanoplasty: a modified technique of cartilage graft tympanoplasty.
Although the temporalis fascia has been widely used since 1957as a graft in tympanoplasty, it can eventually become thin and atrophic. In addition, due to the lack of elasticity and resistance to pressure changes in the external ear canal, secondary perforations may also develop. Therefore, several authors have suggested that temporalis fascia should be strengthened by periosteum or replaced by cartilage. In the present paper the term chondrotympanoplasty is introduced to describe modified techniques of cartilage graft tympanoplasty and intratympanal chondroplasties. These techniques were used in 76 patients who were prospectively followed up for two years post-operatively. In type I chondrotympanoplasty, the mean pre-operative air-bone gap was reduced from 20 dB to 8 dB at 2000 Hz. In 93% of these cases the cartilage graft was taken without problems and there was no perforation in the new eardrum. In two cases of type I chondrotympanoplasty who experienced acute post-operative otitis media, the cartilage graft was found to be relatively resistant to infection, as only a pinhole perforation was noted. Type III chondrotympanoplasty with endotympanic chondroplasty (intratympanal tubal chondroplasty and intratympanal stapes chondroplasty) was found to give good aeration in the cavity, an intact new eardrum, and remarkable hearing results, reducing the mean pre-operative air-bone gap from 25 dB to 10 dB at 2000 Hz. Cartilage was found to be an excellent graft material and the described chondrotympanoplasties gave very promising results. Cartilage should be used more widely in tympanoplasty (chondrotympanoplasty).


Self-association of the amino-terminal domain of the yeast TATA-binding protein.
The amino-terminal domain of yeast TATA-binding protein has been proposed to play a crucial role in the self-association mechanism(s) of the full-length protein. Here we tested the ability of this domain to self-associate under a variety of solution conditions. Escherichia coli two-hybrid assays, in vitro pull-down assays, and in vitro cross-linking provided qualitative evidence for a limited and specific self-association. Sedimentation equilibrium analysis using purified protein was consistent with a monomer-dimer equilibrium with an apparent dissociation constant of approximately 8.4 microM. Higher stoichiometry associations remain possible but could not be detected by any of these methods. These results demonstrate that the minimal structure necessary for amino-terminal domain self-association must be present even in the absence of carboxyl-terminal domain structures. On the basis of these results we propose that amino-terminal domain structures contribute to the oligomerization interface of the full-length yeast TATA-binding protein.


Extravascular fibrin formation and dissolution in synovial tissue of patients with osteoarthritis and rheumatoid arthritis.
Fibrin deposition is a prominent finding in the synovium of patients with rheumatoid arthritis (RA). Macrophages are found in increased numbers in RA synovium, and these cells are known to produce a variety of procoagulant and anticoagulant molecules. Using immunohistologic techniques, the content and distribution of several important components of the coagulation system in the synovium of patients with RA, osteoarthritis (OA), or traumatic joint abnormalities requiring surgery were investigated. Samples from 3 patients from each category were examined in detail. RA synovium (compared with that of patients with OA or joint trauma) had increased numbers of macrophages and increased expression/content of fibrinogen, tissue factor, factor XIII, tissue transglutaminase, cross-linked fibrin (fibrin D dimer), urokinase-type plasminogen activator, and alpha 2-plasmin inhibitor. Macrophage content in RA synovium was increased in both the lining cell areas and the interstitial cell areas. Fibrinogen was distributed throughout the tissue in all samples and was greater in RA synovium. In trauma and OA synovia, tissue factor was seen only in association with vessels (endothelial cells), but in RA synovium, it was markedly increased throughout the tissues. While fibrin D dimer was seen in small amounts in synovial lining cell areas of trauma and OA synovia, it was present in increased amounts in the lining cell and interstitial cell areas of RA synovium. Factor XIII and tissue transglutaminase were present in scant amounts in trauma and OA synovia, but there were increased amounts of both (especially tissue transglutaminase) in RA synovium in the vessel, lining cell, and interstitial cell areas. Urokinase and alpha 2-plasmin inhibitor were also markedly increased in RA synovium. These results suggest that in inflamed synovium, there is ongoing extravascular tissue fibrin formation and dissolution that correlates with the degree of inflammation and macrophage content. Extravascular coagulation/fibrinolysis in RA represents a potential target for therapeutic intervention in this disease.


Community based interventions for the prevention and control of tuberculosis.
In 2012, an estimated 8.6 million people developed tuberculosis (TB) and 1.3 million died from the disease. With its recent resurgence with the human immunodeficiency virus (HIV); TB prevention and management has become further challenging. We systematically evaluated the effectiveness of community based interventions (CBI) for the prevention and treatment of TB and a total of 41 studies were identified for inclusion. Findings suggest that CBI for TB prevention and case detection showed significant increase in TB detection rates (RR: 3.1, 95% CI: 2.92, 3.28) with non-significant impact on TB incidence. CBI for treating patients with active TB showed an overall improvement in treatment success rates (RR: 1.09, 95% CI: 1.07, 1.11) and evidence from a single study suggests significant reduction in relapse rate (RR: 0.26, 95% CI: 0.18, 0.39). The results were consistent for various study design and delivery mechanism. Qualitative synthesis suggests that community based TB treatment delivery through community health workers (CHW) not only improved access and service utilization but also contributed to capacity building and improving the routine TB recording and reporting systems. CBI coupled with the DOTS strategy seem to be an effective approach, however there is a need to evaluate various community-based integrated delivery models for relative effectiveness.


Coordination of the transcriptome and metabolome by the circadian clock.
The circadian clock governs a large array of physiological functions through the transcriptional control of a significant fraction of the genome. Disruption of the clock leads to metabolic disorders, including obesity and diabetes. As food is a potent zeitgeber (ZT) for peripheral clocks, metabolites are implicated as cellular transducers of circadian time for tissues such as the liver. From a comprehensive dataset of over 500 metabolites identified by mass spectrometry, we reveal the coordinate clock-controlled oscillation of many metabolites, including those within the amino acid and carbohydrate metabolic pathways as well as the lipid, nucleotide, and xenobiotic metabolic pathways. Using computational modeling, we present evidence of synergistic nodes between the circadian transcriptome and specific metabolic pathways. Validation of these nodes reveals that diverse metabolic pathways, including the uracil salvage pathway, oscillate in a circadian fashion and in a CLOCK-dependent manner. This integrated map illustrates the coherence within the circadian metabolome, transcriptome, and proteome and how these are connected through specific nodes that operate in concert to achieve metabolic homeostasis.


Cardiac and plasma lipid profiles in response to acute hypoxia in neonatal and young adult rats.
The physiological and biochemical responses to acute hypoxia have not been fully characterized in neonates. Fatty acids and lipids play an important role in most aspects of cardiac function. We performed comprehensive lipid profiling analysis to survey the changes that occur in heart tissue and plasma of neonatal and young adult rats exposed to hypoxia for 2 h, and following 2 h of recovery from hypoxia. Cardiac and plasma concentrations of short-chain acylcarnitines, and most plasma long-chain fatty acids, were decreased in hypoxic neonates. Following recovery from hypoxia, concentrations of propionylcarnitine, palmitoylcarnitine, stearoylcarnitine were increased in neonatal hearts, while oleylcarnitine and linoleylcarnitine concentrations were increased in neonatal plasma. The concentrations of long-chain fatty acids and long-chain acylcarnitines were increased in the hearts and plasma of hypoxic young adult rats; these metabolites returned to baseline values following recovery from hypoxia. There are differential effects of acute hypoxia on cardiac and plasma lipid profiles with maturation from the neonate to the young adult rat. Changes to neonatal cardiac and plasma lipid profiles during hypoxia likely allowed for greater metabolic and physiologic flexibility and increased chances for survival. Persistent alterations in the neonatal cardiac lipid profile following recovery from hypoxia may play a role in the development of rhythm disturbances.


Glycosaminoglycan silencing by engineered CXCL12 variants.
We have engineered GPCR (G protein-coupled receptor) knock-out and high GAG-binding affinity into CXCL12α to inhibit CXCL12α-induced cell migration. Compared to wtCXCL12, the mutant CXCL12α (Δ8 L29K V39K) exhibited a 5.6-fold and a 2.2-fold affinity increase for heparin and heparan sulfate, respectively. From NaCl-based heparin displacement chromatography we concluded that more amino acid replacements would lead to altered GAG (glycosaminoglycan) ligand specificity. GAG silencing by this mutant was shown in a murine seeding model of human cancer cells, whereby a greatly reduced number of liver metastases was detected when the animals were treated intravenously with 1mg/kg CXCL12α (Δ8 L29K V39K) before cancer cell application.


Post-traumatic angle recession glaucoma: a risk factor for bleb failure after trabeculectomy.
In order to determine if post-traumatic angle recession is a risk factor for failure of glaucoma filtering surgery independent of age or race, the surgical results of trabeculectomy performed in 35 consecutive patients with angle recession glaucoma were compared with those of 35 matched patients with primary open angle glaucoma. A postoperative intraocular pressure of < or = 21 mm Hg (with or without glaucoma medication) was found in 15 of the 35 (43%) patients with angle recession glaucoma compared with 26 of the 35 (74%) patients with primary open angle glaucoma. The long term success of trabeculectomy was significantly worse in angle recession glaucoma when the results were analysed using Kaplan-Meier survival curves. Bleb failure occurred a mean period of 3.1 (SD 1.2) months after trabeculectomy in angle recession glaucoma compared with 9.4 (5) months in primary open angle glaucoma (p < or = 0.001). The finding that posttraumatic angle recession is a risk factor for failure of trabeculectomy, supports the use of antimetabolite therapy to suppress fibrosis after trabeculectomy in these patients.


Protective immune response to experimental infection with Sarcocystis neurona in 57BL/6 mice.
Immunocompetent C57BL/6 mice were infected with Sarcocystis neurona merozoites to assess the protective immune response to active infection. Using a direct agglutination test, all infected mice seroconverted to S. neurona merozoite antigens by day 14 postinfection (PI). Further, mice developed splenomegaly and bilateral symmetrical lymphadenopathy by day 14 PI, which appeared to be resolving by day 28 PI. Histologic analysis revealed a marked increase in germinal center formation in the spleen and lymph nodes by day 14 PI. Corresponding to gross and histopathological changes, the percentage of B-cells decreased significantly by day 14 PI but then increased significantly and persisted at day 28 PI in the blood, spleen, and multiple lymph nodes. There was a sharp nonspecific significant decrease in CD4 percentages by day 14 PI in the blood, spleen, and lymph nodes. Early-activation CD8 lymphocytes (CD62/CD8) were significantly down-regulated coinciding with a significant compensatory up-regulation of memory (CD44/CD8) lymphocytes in multiple organs. We propose that the protective cell-mediated immune response to S. neurona involves both CD4 and CD8 cells, with CD8 lymphocytes appearing to play a more critical role.


Calcium-independent inhibitory G-protein signaling induces persistent presynaptic muting of hippocampal synapses.
Adaptive forms of synaptic plasticity that reduce excitatory synaptic transmission in response to prolonged increases in neuronal activity may prevent runaway positive feedback in neuronal circuits. In hippocampal neurons, for example, glutamatergic presynaptic terminals are selectively silenced, creating "mute" synapses, after periods of increased neuronal activity or sustained depolarization. Previous work suggests that cAMP-dependent and proteasome-dependent mechanisms participate in silencing induction by depolarization, but upstream activators are unknown. We, therefore, tested the role of calcium and G-protein signaling in silencing induction in cultured hippocampal neurons. We found that silencing induction by depolarization was not dependent on rises in intracellular calcium, from either extracellular or intracellular sources. Silencing was, however, pertussis toxin sensitive, which suggests that inhibitory G-proteins are recruited. Surprisingly, blocking four common inhibitory G-protein-coupled receptors (GPCRs) (adenosine A(1) receptors, GABA(B) receptors, metabotropic glutamate receptors, and CB(1) cannabinoid receptors) and one ionotropic receptor with metabotropic properties (kainate receptors) failed to prevent depolarization-induced silencing. Activating a subset of these GPCRs (A(1) and GABA(B)) with agonist application induced silencing, however, which supports the hypothesis that G-protein activation is a critical step in silencing. Overall, our results suggest that depolarization activates silencing through an atypical GPCR or through receptor-independent G-protein activation. GPCR agonist-induced silencing exhibited dependence on the ubiquitin-proteasome system, as was shown previously for depolarization-induced silencing, implicating the degradation of vital synaptic proteins in silencing by GPCR activation. These data suggest that presynaptic muting in hippocampal neurons uses a G-protein-dependent but calcium-independent mechanism to depress presynaptic vesicle release.


Associations Between Meeting the 24-Hour Movement Guidelines and Cardiometabolic Risk in Young Children.
The authors aimed to examine the association between meeting the integrative movement behavior guidelines (physical activity, screen viewing, and sleep) and cardiometabolic risk (CMR) factors in young children. In this cross-sectional study, physical activity, screen viewing, and sleep were assessed using parent-reported data. The 24-Hour Movement Guidelines for the Early Years (0-4 y) were defined as 180 minutes of physical activity/day (of which ≥60 min should be moderate-to-vigorous intensity), ≤1 hour of screen viewing/day, and 10 to 13 hours of sleep/night. Waist circumference, glucose, high-density lipoprotein cholesterol, triglycerides, and systolic blood pressure were measured in a clinical setting by trained staff. A total CMR score and individual CMR factors served as primary and secondary outcomes, respectively. Of the 767 participants (3-4 y), 26.4% met none of the guideline's recommendations, whereas 41.3%, 33.1%, and 10.6% of the sample met 1, 2, or all 3 recommendations, respectively. The number of recommendations met was not associated with the total CMR score or individual CMR factors (P > .05), with the exceptions of high-density lipoprotein (odds ratio = 1.61; 95% confidence interval, 1.11 to 2.33; P = .01). Meeting the 24-Hour Movement Guidelines in early childhood was not associated with overall CMR, but was associated with favorable cholesterol outcomes.


My burning issues in the neoadjuvant treatment for breast cancer.
A combination of anthracyclines and taxanes remains the standard of care for neoadjuvant chemotherapy (NACT) resulting in increased breast conservation rate (BCR) and decreased recurrence rate [1]. Whether pathological complete response (pCR) could be an appropriate surrogate parameter for long-term survival is still a matter of debate. In patients with triple-negative breast cancer (TNBC) and HER2-positive breast cancer (BC), a six to nine times higher risk for relapse has been reported if no pCR was achieved [2, 3]. Within these aggressive subtypes the strongest association between pCR and long-term outcome could be observed [4]. However, a pooled analysis of recently conducted trials could only identify pCR as a surrogate endpoint for improved event-free survival (EFS) and overall survival (OS) on an individual patient level as opposed to the trial level [5]. Even in TNBC, demonstrating that an increased pCR converts into a significant survival benefit would require a study population markedly larger than calculated for previously conducted trials [6, 7].


Release of human cytomegalovirus from latency by a KAP1/TRIM28 phosphorylation switch.
Human cytomegalovirus (HCMV) is a highly prevalent pathogen that induces life-long infections notably through the establishment of latency in hematopoietic stem cells (HSC). Bouts of reactivation are normally controlled by the immune system, but can be fatal in immuno-compromised individuals such as organ transplant recipients. Here, we reveal that HCMV latency in human CD34(+) HSC reflects the recruitment on the viral genome of KAP1, a master co-repressor, together with HP1 and the SETDB1 histone methyltransferase, which results in transcriptional silencing. During lytic infection, KAP1 is still associated with the viral genome, but its heterochromatin-inducing activity is suppressed by mTOR-mediated phosphorylation. Correspondingly, HCMV can be forced out of latency by KAP1 knockdown or pharmacological induction of KAP1 phosphorylation, and this process can be potentiated by activating NFkB with TNF-α. These results suggest new approaches both to curtail CMV infection and to purge the virus from organ transplants.


A family with erythrocytosis establishes a role for prolyl hydroxylase domain protein 2 in oxygen homeostasis.
The number of red blood cells is normally tightly regulated by a classic homeostatic mechanism based on oxygen sensing in the kidney. Decreased oxygen delivery resulting from anemia induces the production of erythropoietin, which increases red cell production and hence oxygen delivery. Investigations of erythropoietin regulation identified the transcription factor hypoxia-inducible factor (HIF). HIF is now recognized as being a key regulator of genes that function in a comprehensive range of processes besides erythropoiesis, including energy metabolism and angiogenesis. HIF itself is regulated through the alpha-subunit, which is hydroxylated in the presence of oxygen by a family of three prolyl hydroxylase domain proteins (PHDs)/HIF prolyl hydroxylases/egg-laying-defective nine enzymes. Hydroxylation allows capture by the von Hippel-Lindau tumor suppressor gene product, ubiquitination, and destruction by the proteasome. Here we describe an inherited mutation in a mammalian PHD enzyme. We show that this mutation in PHD2 results in a marked decrease in enzyme activity and is associated with familial erythrocytosis, identifying a previously unrecognized cause of this condition. Our findings indicate that PHD2 is critical for normal regulation of HIF in humans.


Copper-related blood indexes in kidney dialysis patients.
Previous work has suggested that kidney hemodialysis patients could be at risk for either moderate copper deficiency or copper toxicity. The present study examined copper-related blood indexes in subjects undergoing hemodialysis treatments with membranes that are not copper-based, in subjects undergoing chronic ambulatory peritoneal dialysis (CAPD), and in control subjects. Both dialysis groups had low plasma copper and ceruloplasmin activities. This occurred despite high plasma interleukin 6 concentrations, a situation that usually elevates plasma ceruloplasmin and copper values. CAPD and hemodialysis subjects had low ratios of ceruloplasmin activity to immunoreactive protein, and low ratios of plasma copper to ceruloplasmin protein. Both are signs of copper deficiency. In contrast, copper-containing erythrocyte superoxide dismutase (SOD) activities were high in hemodialysis subjects and showed a nonsignificant trend toward high values in CAPD subjects. Blood mononuclear cell copper contents were highly variable within each group, and there were no significant differences between groups. In conclusion, ceruloplasmin-related indexes in kidney dialysis patients not dialyzed with copper-based membranes suggested a tendency toward moderate copper deficiency. However, this contention could not be confirmed by erythrocyte SOD activity or mononuclear cell copper measurements.


A rare but severe pulmonary side effect of cetuximab in two patients.
Cetuximab is a monoclonal antibody that treats malignant disease by inhibiting epidermal growth factor receptor. Many common adverse events have been reported and include skin rashes, infusion reactions, gastrointestinal complaints and headache. Up to now, cetuximab-associated pulmonary toxicity has been rarely reported in the literature. The present report describes two cases with probable interstitial pneumonitis 5-6 weeks after commencing treatment with chemotherapy combined with cetuximab. One patient recovered and the second patient died due to gastrointestinal bleeding 3 weeks after an initial response to steroids.


SORTA: a system for ontology-based re-coding and technical annotation of biomedical phenotype data.
There is an urgent need to standardize the semantics of biomedical data values, such as phenotypes, to enable comparative and integrative analyses. However, it is unlikely that all studies will use the same data collection protocols. As a result, retrospective standardization is often required, which involves matching of original (unstructured or locally coded) data to widely used coding or ontology systems such as SNOMED CT (clinical terms), ICD-10 (International Classification of Disease) and HPO (Human Phenotype Ontology). This data curation process is usually a time-consuming process performed by a human expert. To help mechanize this process, we have developed SORTA, a computer-aided system for rapidly encoding free text or locally coded values to a formal coding system or ontology. SORTA matches original data values (uploaded in semicolon delimited format) to a target coding system (uploaded in Excel spreadsheet, OWL ontology web language or OBO open biomedical ontologies format). It then semi- automatically shortlists candidate codes for each data value using Lucene and n-gram based matching algorithms, and can also learn from matches chosen by human experts. We evaluated SORTA's applicability in two use cases. For the LifeLines biobank, we used SORTA to recode 90 000 free text values (including 5211 unique values) about physical exercise to MET (Metabolic Equivalent of Task) codes. For the CINEAS clinical symptom coding system, we used SORTA to map to HPO, enriching HPO when necessary (315 terms matched so far). Out of the shortlists at rank 1, we found a precision/recall of 0.97/0.98 in LifeLines and of 0.58/0.45 in CINEAS. More importantly, users found the tool both a major time saver and a quality improvement because SORTA reduced the chances of human mistakes. Thus, SORTA can dramatically ease data (re)coding tasks and we believe it will prove useful for many more projects. Database URL: http://molgenis.org/sorta or as an open source download from http://www.molgenis.org/wiki/SORTA.


Er,Cr:YSGG Laser Performance Improves Biological Response on Titanium Surfaces.
Porphyromonas gingivalis infection is one of the causes of implant failures, which can lead to peri-implantitis. Implant surface roughness is reportedly related strongly to P. gingivalis adhesion, which can lead to peri-implantitis and, later, cell adhesion. Our aim was to evaluate the effects of Er,Cr:YSGG laser on titanium (Ti) disc surfaces and its interaction with bacterial adhesion and fibroblast viability. Ti discs underwent two treatments: autoclaving (control) and erbium, chromium-doped yttrium scandium gallium garnet (Er,Cr:YSGG) laser treatment (test). Ti disc surfaces were examined with scanning electronic microscope (SEM), Energy-dispersive spectrometry (EDX), X-ray photoelectron spectroscopy (XPS). The surface roughness same as wettability were also investigated. Fibroblast viability was assessed with the water-soluble tetrazolium 1 (WST-1) test, and osteoblast differentiation was assessed with the alkaline phosphatase (ALP) assay. Bacterial structure and colony formation were detected with scanning electron microscopy and Gram stain. In comparison to control discs, the test discs showed smoother surfaces, with 0.25-µm decrease in surface roughness (p < 0.05); lower P. gingivalis adhesion (p < 0.01); less P. gingivalis colonization (p < 0.05); and increased fibroblast viability and osteoblast differentiation (p < 0.05). Er,Cr:YSGG laser treatment improved disc surfaces by making them slightly smoother, which reduced P. gingivalis adhesion and increased fibroblast viability and osteoblast differentiation. Er,Cr:YSGG laser treatment can be considered a good option for managing peri-implantitis. Further investigations of laser-assisted therapy are necessary for better guidelines in the treatment of peri-implantitis.


Tyrosine kinome sequencing of pediatric acute lymphoblastic leukemia: a report from the Children's Oncology Group TARGET Project.
One recently identified subtype of pediatric B-precursor acute lymphoblastic leukemia (ALL) has been termed BCR-ABL1-like or Ph-like because of similarity of the gene expression profile to BCR-ABL1 positive ALL suggesting the presence of lesions activating tyrosine kinases, frequent alteration of IKZF1, and poor outcome. Prior studies demonstrated that approximately half of these patients had genomic lesions leading to CRLF2 overexpression, with half of such cases harboring somatic mutations in the Janus kinases JAK1 and JAK2. To determine whether mutations in other tyrosine kinases might also occur in ALL, we sequenced the tyrosine kinome and downstream signaling genes in 45 high-risk pediatric ALL cases with either a Ph-like gene expression profile or other alterations suggestive of activated kinase signaling. Aside from JAK mutations and 1 FLT3 mutation, no somatic mutations were found in any other tyrosine kinases, suggesting that alternative mechanisms are responsible for activated kinase signaling in high-risk ALL.


Verification of Implant Surface Modification by a Novel Processing Method.
Metals have been used clinically as biomaterials, especially in the orthopaedic and dental fields. Metals used as implants wear at contact surfaces, producing metal particles and metal ions that may be harmful. Newly developed metal implants and methods of implant surface modification are currently under scrutiny. We evaluated the use of electrolytic in-process dressing (ELID) as a surface finishing method for metal implants. Metal implants processed using the ELID method (ELID group) or not processed (Non-ELID group) were inserted surgically into rabbit femurs. The rabbits were sacrificed postoperatively over a 24-week period. We assessed the concentrations of the cytokines, interleukin (IL)-1β, IL-6, and tumor necrosis factor-α, the resistance to implant pull-out, and histopathology at the implant site. There was no significant difference between the groups regarding the cytokine concentrations or implant pull-out resistance. Many particles indicating wear around the implant were noted in the Non-ELID group (n=10) but not the ELID group (n=13), while a fibrous membrane adhering to the every implant was noted in the ELID group. The formation of a fibrous membrane rather than metal particles in the ELID group may indicate improved biocompatibility, and it suggests that ELID may prevent corrosion in the areas of contact.


Isolating plasma from blood using a dielectrophoresis-active hydrophoretic device.
Plasma is a complex substance that contains proteins and circulating nucleic acids and viruses that can be utilised for clinical diagnostics, albeit a precise analysis depends on the plasma being totally free of cells. We proposed the use of a dielectrophoresis (DEP)-active hydrophoretic method to isolate plasma from blood in a high-throughput manner. This microfluidic device consists of anisotropic microstructures embedded on the top of the channel which generate lateral pressure gradients while interdigitised electrodes lay on the bottom of the channel which can push particles or cells into a higher level using a negative DEP force. Large and small particles or cells (3 μm and 10 μm particles, and red blood cells, white blood cells, and platelets) can be focused at the same time in our DEP-active hydrophoretic device at an appropriate flow rate and applied voltage. Based on this principle, all the blood cells were filtrated from whole blood and then the plasma was extracted with a purity of 94.2% and a yield of 16.5% at a flow rate of 10 μL min(-1). This solved the challenging problem caused by the relatively low throughput of the DEP based device. Our DEP-active hydrophoretic device is a flexible and tunable system that can control the lateral positions of particles by modulating the external voltages without redesigning and fabricating a new channel, and because it is easy to operate, it is easily compatible with other microfluidic platforms that are used for further detection.


Characterization of immunoglobulin G antibodies to Plasmodium falciparum sporozoite surface antigen MB2 in malaria exposed individuals.
MB2 protein is a sporozoite surface antigen on the human malaria parasite Plasmodium falciparum. MB2 was identified by screening a P. falciparum sporozoite cDNA expression library using immune sera from a protected donor immunized via the bites of P. falciparum-infected irradiated mosquitoes. It is not known whether natural exposure to P. falciparum also induces the anti-MB2 response and if this response differs from that in protected individuals immunized via the bites of P. falciparum infected irradiated mosquitoes. The anti-MB2 antibody response may be part of a robust protective response against the sporozoite. Fragments of polypeptide regions of MB2 were constructed as recombinant fusions sandwiched between glutathione S-transferase and a hexa histidine tag for bacterial expression. The hexa histidine tag affinity purified proteins were used to immunize rabbits and the polyclonal sera evaluated in an in vitro inhibition of sporozoite invasion assay. The proteins were also used in immunoblots with sera from a limited number of donors immunized via the bites of P. falciparum infected irradiated mosquitoes and plasma and serum obtained from naturally exposed individuals in Kenya. Rabbit polyclonal antibodies targeting the non-repeat region of the basic domain of MB2 inhibited sporozoites entry into HepG2-A16 cells in vitro. Analysis of serum from five human volunteers that were immunized via the bites of P. falciparum infected irradiated mosquitoes that developed immunity and were completely protected against subsequent challenge with non-irradiated parasite also had detectable levels of antibody against MB2 basic domain. In contrast, in three volunteers not protected, anti-MB2 antibodies were below the level of detection. Sera from protected volunteers preferentially recognized a non-repeat region of the basic domain of MB2, whereas plasma from naturally-infected individuals also had antibodies that recognize regions of MB2 that contain a repeat motif in immunoblots. Sequence analysis of eleven field isolates and four laboratory strains showed that these antigenic regions of the basic domain of the MB2 gene are highly conserved in parasites obtained from different parts of the world. Moreover, anti-MB2 antibodies also were detected in the plasma of 83% of the individuals living in a malaria endemic area of Kenya (n = 41). A preliminary analysis of the human humoral response against MB2 indicates that it may be an additional highly conserved target for immune intervention at the pre-erythrocytic stage of P. falciparum life cycle.


Revisiting Warfarin Dosing Using Machine Learning Techniques.
Determining the appropriate dosage of warfarin is an important yet challenging task. Several prediction models have been proposed to estimate a therapeutic dose for patients. The models are either clinical models which contain clinical and demographic variables or pharmacogenetic models which additionally contain the genetic variables. In this paper, a new methodology for warfarin dosing is proposed. The patients are initially classified into two classes. The first class contains patients who require doses of >30 mg/wk and the second class contains patients who require doses of ≤30 mg/wk. This phase is performed using relevance vector machines. In the second phase, the optimal dose for each patient is predicted by two clinical regression models that are customized for each class of patients. The prediction accuracy of the model was 11.6 in terms of root mean squared error (RMSE) and 8.4 in terms of mean absolute error (MAE). This was 15% and 5% lower than IWPC and Gage models (which are the most widely used models in practice), respectively, in terms of RMSE. In addition, the proposed model was compared with fixed-dose approach of 35 mg/wk, and the model proposed by Sharabiani et al. and its outperformance were proved in terms of both MAE and RMSE.


Predicting the potential distribution and forest impact of the invasive species Cydalima perspectalis in Europe.
Invasive species have considerably increased in recent decades due to direct and indirect effects of ever-increasing international trade rates and new climate conditions derived from global change. We need to better understand how the dynamics of early species invasions develop and how these result in impacts on the invaded ecosystems. Here we studied the distribution and severe defoliation processes of the box tree moth (Cydalima perspectalis W.), a tree defoliator insect native to Asia and invasive in Europe since 2007, through the combination of species distribution models based on climate and landscape composition information. The results showed that the combination of data from the native and the invaded areas was the most effective methodology for the appropriate invasive species modeling. The species was not influenced by overall landscape factors, but only by the presence of its host plant, dispersal capacity, and climate suitability. Such climate suitability was described by low precipitation seasonality and minimum annual temperatures around 0°C, defining a continentality effect throughout the territory. We emphasize the need of studying distribution and severe defoliation processes separately because we identified that climate suitability was slightly involved in limiting species spread processes but strongly constrained ecosystem impact in terms of defoliation before the species reaches equilibrium with the new environment. New studies on habitat recovery after disturbance, ecological consequences of such impact, and community dynamics in a context of climate change are required for a better understanding of this invasive species.


Expression of the floral repressor miRNA156 is positively regulated by the AGAMOUS-like proteins AGL15 and AGL18.
The regulation of flowering time has crucial implications for plant fitness. MicroRNA156 (miR156) represses the floral transition in Arabidopsis thaliana, but the mechanisms regulating its transcription remain unclear. Here, we show that two AGAMOUS-like proteins, AGL15 and AGL18, act as positive regulators of the expression of MIR156. Small RNA northern blot analysis revealed a significant decrease in the levels of mature miR156 in agl15 agl18 double mutants, but not in the single mutants, suggesting that AGL15 and AGL18 co-regulate miR156 expression. Histochemical analysis further indicated that the double mutants showed a reduction in MIR156 promoter strength. The double mutants also showed reduced abundance of pri-miR156a and pri-miR156c, two of the primary transcripts from MIR156 genes. Electrophoretic mobility shift assays demonstrated that AGL15 directly associated with the CArG motifs in the MIR156a/c promoters. AGL18 did not show binding affinity to the CArG motifs, but pull-down and yeast two-hybrid assays showed that AGL18 forms a heterodimer with AGL15. GFP reporter assays and bimolecular fluorescence complementation (BiFC) showed that AGL15 and AGL18 co-localize in the nucleus and confirmed their in vivo interaction. Overexpression of miR156 did not affect the levels of AGL15 and AGL18 transcripts. Taking these data together, we present a model for the transcriptional regulation of MIR156. In this model, AGL15 and AGL18 may form a complex along with other proteins, and bind to the CArG motifs of the promoters of MIR156 to activate the MIR156 expression.


Characterization of Plasmodium falciparum calcium-dependent protein kinase 1 (PfCDPK1) and its role in microneme secretion during erythrocyte invasion.
Calcium-dependent protein kinases (CDPKs) play important roles in the life cycle of Plasmodium falciparum and other apicomplexan parasites. CDPKs commonly have an N-terminal kinase domain (KD) and a C-terminal calmodulin-like domain (CamLD) with calcium-binding EF hands. The KD and CamLD are separated by a junction domain (JD). Previous studies on Plasmodium and Toxoplasma CDPKs suggest a role for the JD and CamLD in the regulation of kinase activity. Here, we provide direct evidence for the binding of the CamLD with the P3 region (Leu(356) to Thr(370)) of the JD in the presence of calcium (Ca(2+)). Moreover, site-directed mutagenesis of conserved hydrophobic residues in the JD (F363A/I364A, L356A, and F350A) abrogates functional activity of PfCDPK1, demonstrating the importance of these residues in PfCDPK1 function. Modeling studies suggest that these residues play a role in interaction of the CamLD with the JD. The P3 peptide, which specifically inhibits the functional activity of PfCDPK1, blocks microneme discharge and erythrocyte invasion by P. falciparum merozoites. Purfalcamine, a previously identified specific inhibitor of PfCDPK1, also inhibits microneme discharge and erythrocyte invasion, confirming a role for PfCDPK1 in this process. These studies validate PfCDPK1 as a target for drug development and demonstrate that interfering with its mechanistic regulation may provide a novel approach to design-specific PfCDPK1 inhibitors that limit blood stage parasite growth and clear malaria parasite infections.


Adaptation and heterogeneity of Escherichia coli MC1000 growing in complex environments.
In a study aiming to assess bacterial evolution in complex growth media, we evaluated the long-term adaptive response of Escherichia coli MC1000 in Luria-Bertani (LB) medium. Seven parallel populations were founded and followed over 150 days in sequential batch cultures under three different oxygen conditions (defined environments), and 19 evolved forms were isolated. The emergence of forms with enhanced fitness was evident in competition experiments of all evolved forms versus the ancestral strain. The evolved forms were then subjected to phenotypic and genomic analyses relative to the ancestor. Profound changes were found in their phenotypes as well as whole-genome sequences. Interestingly, considerable heterogeneity was found at the intrapopulational level. However, consistently occurring parallel adaptive responses were found across all populations. The evolved forms all contained a mutation in galR, a repressor of the galactose operon. Concomitantly, the new forms revealed enhanced growth on galactose as well as galactose-containing disaccharides. This response was likely driven by the LB medium.


Towards the design of an antibody that recognises a given protein epitope.
We have explored the possibility of designing repertoires of antibodies complementary to a given protein epitope, specifically the face of the ribonuclease inhibitor barstar that binds to the enzyme barnase. An antibody repertoire was created by mutation of ten residues in the hypervariable loops of a synthetic antibody fragment and displayed on filamentous bacteriophage. The positions of three of the ten residues of the antibody (VL 32, 50 and 94) were chosen to match a triangle of three negative charges on the face of barstar and mutated to favour residues of opposite charge or those with hydrogen-bonding potential. The other seven residues, chosen to allow for variation in the surface of interaction, were mutated at random. One of the antibody fragments isolated after selection of the repertoire (10(8) clones per library) was shown to bind to barstar with an affinity of 1.0x10(-7) M and the binding was competed by barnase. Furthermore, the binding of the antibody to barstar was highly sensitive to mutation of any of five residues of barstar known to contact barnase. This indicates that it may be possible, by a combination of design and selection, to build antibodies to a given epitope.


A study of phonomicrosurgical arm support postures using a magnetic motion tracking system.
To study the different arm support postures used in phonomicrosurgery by using a magnetic-based phonomicrosurgery instrument tracking system (MPTS). Through quantitative motion parameter data collected from four arm support postures (elbow support [ES], forearm support [FS], forearm and hand support, and no support), phonomicrosurgical operation postures were analyzed and compared. Prospective cohort study. Seven subjects operated on phonomicrosurgical simulation cutting tasks with four arm support postures while being monitored by MPTS. The motion parameters, including operation time, path length, depth perception, and motion smoothness were analyzed. The subjects' cutting quality was also calculated. With the FS, the nondominant hand showed improved S, better D, and shorter P (P < 0.05). Better motion control in the dominant hand resulted from ES posture (P < 0.05). Better operation quality was associated with increased motion control in the nondominant hand. Forearm support resulted in higher steadiness and shorter surgical path in the nondominant hand. In the dominant hand, ES resulted in increased steadiness, shorter surgical path, and better D. The effect of both gravity and wrist dexterity on movement control should be considered when selecting proper arm supports. 4.


Nimesulide, a selective COX-2 inhibitor, acts synergistically with ionizing radiation against A549 human lung cancer cells through the activation of caspase-8 and caspase-3.
Several lines of evidence suggest that non-steroidal anti-inflammatory drugs (NSAIDs) have a radiosensitizing effect on cancer cells in vitro and in vivo, but little is known about the underlying cellular mechanism. In this study, we found that the treatment with the NSAID nimesulide significantly increased the sensitivity of A549 human non-small cell lung cancer cells to radiotherapy. The combined nimesulide-radiation treatment increased apoptosis, induced the cleavage of caspase-3, caspase-9, and poly(ADP-ribose) polymerase (PARP), activated caspase-8, and induced cleavage of Bid. A pan-caspase inhibitor, z-VAD-fmk, suppressed this increase in apoptosis and also suppressed the cleavage of caspase-8, caspase-3, and PARP, suggesting a caspase-dependent mechanism. In addition, z-IETD-fmk, a selective caspase-8 inhibitor, suppressed the nimesulide- and radiation-induced cleavage activation of caspase-9, caspase-3, caspase-8, and Bid, and suppressed the concomitant apoptosis, indicating that the nimesulide-induced increase in radiosensitivity was initiated by caspase-8. However, the caspase-3 inhibitor z-DEVD-fmk failed to suppress activation of the caspase-8/Bid pathway, indicating that caspase-3 activation occurred downstream of caspase-8 activation in our experiments. Marked antitumor effects, which were evaluated by measuring protracted tumor regression, were observed when nude mice were treated with a combination of nimesulide at a clinically achievable dose (0.5 mg/kg) and radiation therapy. Our results, demonstrating the radiosensitivity-increasing and tumor growth-inhibiting effects of nimesulide, suggest that nimesulide may be suitable as an adjuvant to enhance the efficacy and selectivity of radiotherapy.


Use of sequential extraction to assess the influence of sewage sludge amendment on metal mobility in Chilean soils.
In Chile, the increasing number of plants for the treatment of wastewater has brought about an increase in the generation of sludge. One way of sludge disposal is its application on land; this, however involves some problems, some of them being heavy metal accumulation and the increase in organic matter and other components from sewage sludge which may change the distribution and mobility of heavy metals. The purpose of the present study was to determine the effect of sewage sludge application on the distribution of Cr, Ni, Cu, Zn and Pb in agricultural soils in Chile. Three different soils, two Mollisols and one Alfisol, were sampled from an agricultural area in Central Chile. The soils were treated with sewage sludge at the rates of 0 and 30 ton ha(-1), and were incubated at 25 degrees C for 45 days. Before and after incubation, the soils were sequentially extracted to obtain labile (exchangeable and sodium acetate-soluble), potentially labile (soluble in moderately reducing conditions, K4P2O7-soluble and soluble in reducing conditions) and inert (soluble in strong acid oxidizing conditions) fractions. A two-level factored design was used to assess the effect of sludge application rate, incubation time and their interaction on the mobility of the elements under study. Among the metals determined in the sludge, zinc has the highest concentration. However, with the exception of Ni, the total content of metals was lower than the recommended limit values in sewage sludge as stated by Chilean regulations. Although 23% of zinc in sludge was in more mobile forms, the residual fraction of all metals was the predominant form in soils and sludge. The content of zinc only was significantly increased in two of the soils by sewage sludge application. On the other hand, with the exception of copper, the metals were redistributed in the first four fractions of amended soils. The effect of sludge application rate, incubation time and their interaction depended on the metal or soil type. In most cases an increase in more mobile forms of metals in soils was observed as the final effect.


Innate and Adaptive Immune Responses in Chronic HCV Infection.
Hepatitis C virus (HCV) remains a public health problem of global importance, even in the era of potent directly-acting antiviral drugs. In this chapter, I discuss immune responses to acute and chronic HCV infection. The outcome of HCV infection is influenced by viral strategies that limit or delay the initiation of innate antiviral responses. This delay may enable HCV to establish widespread infection long before the host mounts effective T and B cell responses. HCV's genetic agility, resulting from its high rate of replication and its error prone replication mechanism, enables it to evade immune recognition. Adaptive immune responses fail to keep up with changing viral epitopes. Neutralizing antibody epitopes may be hidden by decoy structures, glycans, and lipoproteins. T cell responses fail due to changing epitope sequences and due to exhaustion, a phenomenon that may have evolved to limit immune-mediated pathology. Despite these difficulties, innate and adaptive immune mechanisms do impact HCV replication. Immune-mediated clearance of infection is possible, occurring in 20-50% of people who contract the disease. New developments raise hopes for effective immunological interventions to prevent or treat HCV infection.


A Pilot Study of Four Intraosseous Blood Transfusion Strategies.
Intraosseous (IO) access is used by military first responders administering fluids, blood, and medications. Current IO transfusion strategies include gravity, pressure bags, rapid transfusion devices, and manual push-pull through a three-way stopcock. In a swine model of hemorrhagic shock, we compared flow rates among four different IO blood transfusion strategies. Nine Yorkshire swine were placed under general anesthesia. We removed 20 to 25mL/kg of each animal's estimated blood volume using flow of gravity. IO access was obtained in the proximal humerus. We then autologously infused 10 to 15mL/kg of the animal's estimated blood volume through one of four randomly assigned treatment arms. The average weight of the swine was 77.3kg (interquartile range, 72.7kg-88.8kg). Infusion rates were as follows: gravity, 5mL/min; Belmont rapid infuser, 31mL/min; single-site pressure bag, 78mL/min; double-site pressure bag, 103mL/min; and push-pull technique, 109mL/min. No pulmonary arterial fat emboli were noted. The optimal IO transfusion strategy for injured Servicemembers appears to be single-site transfusion with a 10mL to 20mL flush of normal saline, followed immediately by transfusion under a pressure bag. Further study, powered to detect differences in flow rate and clinical complications. is required.


Exploring the Benefits of Proactive Participation among Adults and Older People by Writing Blogs.
The use of Information and Communication Technologies (ICTs) for information-seeking, social contact and leisure activities is increasing in adults and older people. However, little is known about adults and older people who are already actively using ICTs to write a blog. The aim of this paper is to describe the benefits adults and older people gain from having a blog. Twenty-three older adult bloggers (aged 60-83 years; most of them with college degrees and retired) from Spain who, at the time of the study had an active blog were interviewed. A thematic analysis identified four different benefits related to blogging: (1) a general sense of satisfaction from producing the blog; (2) relational benefits; (3) cognitive benefits; and (4) identity benefits. Results showed that adults and older people experienced a variety of benefits that broadens the distinction between personal and social benefits found in previous research. Blogging in later life challenges the traditional passive/consumer and online user experience view of adults and older people and seems to be a good example of proactive participation through websites.


Effect of sulfur content in wet or dry distillers grains fed at several inclusions on cattle growth performance, ruminal parameters, and hydrogen sulfide.
Effects of S from wet or dry distillers grains with solubles (DGS) containing 0.82 or 1.16% S on animal growth performance, carcass characteristics, apparent total tract digestibility, and ruminal parameters were evaluated. In Exp. 1, crossbred beef steers (n = 120; 345 ± 34 kg BW) were individually fed ad libitum using Calan gates. Treatments were applied as a 2 × 2 × 3 + 1 factorial treatment arrangement with factors of DGS type (wet or dry), S content in DGS (0.82 or 1.16% DM basis), and DGS inclusion (20, 30, and 40%, DM basis), as well as a corn control diet (no DGS). In Exp. 2, ruminally cannulated crossbred beef steers (n = 6; 381 ± 31 kg BW) were assigned to 1 of 5 diets in a 5 × 6 unbalanced Latin Square design and fed ad libitum through five 14-d periods. A 2 × 2 + 1 factorial treatment arrangement was used with the factors of DGS type and S content in DGS (similar to Exp. 1). Inclusion of DGS was 40%, except for a MATCH diet containing wet 1.16% S DGS included at 31.4% (DM basis). Intake of DM decreased linearly (P < 0.01) and quadratically (P < 0.01) for steers fed wet and dry DGS that was 1.16% S, respectively. In addition, steers fed dry DGS consumed 9% more DM (P < 0.01) than those fed wet. Gain decreased linearly (P = 0.02) when wet 1.16% S DGS increased in the diet, representing a 12% drop in ADG between the Control and 40% DGS inclusion. A quadratic (P = 0.02) improvement in G:F was observed for steers fed wet DGS compared with dry, regardless of S content (P = 0.52). Feeding diets with wet 1.16% S DGS linearly decreased (P = 0.03) HCW. In Exp. 2, molar proportion of propionate declined (P = 0.01) 9% and A:P ratio tended (P = 0.13) to be greater when 1.16 compared with 0.82% S DGS was fed. Apparent total tract DMD was not affected (P > 0.16) and only subtle changes (P < 0.01) in ruminal pH parameters were observed. Greater (P = 0.02) ruminal H2S concentration for steers fed wet compared with dry DGS was observed, while 1.16% S DGS tended (P = 0.12) to produce greater ruminal H2S than 0.82% S. Sulfur in wet DGS appears to be more prone to be converted to ruminal H2S, because feeding 1.16% S as wet DGS had a greater impact on ADG, DMI, and ruminal H2S compared with dry DGS.


Integrative multi-omic analysis identifies new drivers and pathways in molecularly distinct subtypes of ALS.
Amyotrophic lateral sclerosis (ALS) is an incurable and fatal neurodegenerative disease. Increasing the chances of success for future clinical strategies requires more in-depth knowledge of the molecular basis underlying disease heterogeneity. We recently laid the foundation for a molecular taxonomy of ALS by whole-genome expression profiling of motor cortex from sporadic ALS (SALS) patients. Here, we analyzed copy number variants (CNVs) occurring in the same patients, by using a customized exon-centered comparative genomic hybridization array (aCGH) covering a large panel of ALS-related genes. A large number of novel and known disease-associated CNVs were detected in SALS samples, including several subgroup-specific loci, suggestive of a great divergence of two subgroups at the molecular level. Integrative analysis of copy number profiles with their associated transcriptomic data revealed subtype-specific genomic perturbations and candidate driver genes positively correlated with transcriptional signatures, suggesting a strong interaction between genomic and transcriptomic events in ALS pathogenesis. The functional analysis confirmed our previous pathway-based characterization of SALS subtypes and identified 24 potential candidates for genomic-based patient stratification. To our knowledge, this is the first comprehensive "omics" analysis of molecular events characterizing SALS pathology, providing a road map to facilitate genome-guided personalized diagnosis and treatments for this devastating disease.


Methodological issues in the design of a rheumatoid arthritis activity score and its cut-offs.
Activity of rheumatoid arthritis (RA) can be evaluated using several scoring scales based on clinical features. The most widely used one is the Disease Activity Score involving 28 joint counts (DAS28) for which cut-offs were proposed to help physicians classify patients. However, inaccurate scoring can lead to inappropriate medical decisions. In this article some methodological issues in the design of such a score and its cut-offs are highlighted in order to further propose a strategy to overcome them. As long as the issues reviewed in this article are not addressed, results of studies based on standard disease activity scores such as DAS28 should be considered with caution.


GANDivAWeb: a web server for detecting early folding units ("foldons") from protein 3D structures.
It has long been known that small regions of proteins tend to fold independently and are then stabilized by interactions between these distinct subunits or modules. Such units, also known as autonomous folding units (AFUs) or"foldons" play a key role in protein folding. A knowledge of such early folding units has diverse applications in protein engineering as well as in developing an understanding of the protein folding process. Such AFUs can also be used as model systems in order to study the structural organization of proteins. In an earlier work, we had utilized a global network partitioning algorithm to identify modules in proteins. We had shown that these modules correlate well with AFUs. In this work, we have developed a webserver, GANDivAWeb, to identify early folding units or "foldons" in networks using the algorithm described earlier. The website has three functionalities: (a) It is able to display information on the modularity of a database of 1420 proteins used in the original work, (b) It can take as input an uploaded PDB file, identify the modules using the GANDivA algorithm and email the results back to the user and (c) It can take as input an uploaded PDB file and a results file (obtained from functionality (b)) and display the results using the embedded viewer. The results include the module decomposition of the protein, plots of cartoon representations of the protein colored by module identity and connectivity as well as contour plots of the hydrophobicity and relative accessible surface area (RASA) distributions. We believe that the GANDivAWeb server, will be a useful tool for scientists interested in the phenomena of protein folding as well as in protein engineering. Our tool not only provides a knowledge of the AFUs through a natural graph partitioning approach but is also able to identify residues that are critical during folding. It is our intention to use this tool to study the topological determinants of protein folding by analyzing the topological changes in proteins over the unfolding/folding pathways.


Pharmacological plus optical penalization treatment for amblyopia: results of a randomized trial.
To compare weekend atropine sulfate use augmented by a plano lens for the sound eye with weekend atropine use alone for moderate amblyopia in children aged 3 years to younger than 7 years. In a multicenter clinical trial, 180 children with moderate amblyopia (visual acuities of 20/40-20/100) were randomized to weekend atropine use augmented by a plano lens or weekend atropine use alone. Masked assessment of amblyopic eye visual acuity using the Amblyopia Treatment Study HOTV testing protocol at 18 weeks. At 18 weeks, amblyopic eye improvement averaged 2.8 lines in the group that received atropine plus a plano lens and 2.4 lines in the group that received atropine alone (mean difference between groups adjusted for baseline acuity, 0.3 line; 95% confidence interval, -0.2 to 0.8 line). Amblyopic eye visual acuity was 20/25 or better in 24 patients (29%) in the group that received atropine only and 35 patients (40%) in the group that received atropine plus a plano lens (P = .03). More patients in the group that received atropine plus a plano lens had reduced sound eye visual acuity at 18 weeks; however, there were no cases of persistent reverse amblyopia. As an initial treatment for moderate amblyopia, the augmentation of weekend atropine use with a plano lens does not substantially improve amblyopic eye visual acuity when compared with weekend atropine use alone. Application to Clinical Practice Treatment of children with unilateral amblyopia. Trial Registration clinicaltrials.org Identifier: NCT00315302.


Design and synthesis of 2-substituted-5-(4-trifluoromethylphenyl-sulphonamido)benzoxazole derivatives as human GST P1-1 inhibitors.
The glutathione transferases (GSTs) are a family of widely distributed Phase II detoxification enzymes. GST P1-1 is frequently overexpressed in rat and human tumours. It is suggested that overexpression of hGST P1-1 by human tumor cells may play a role in resistance to cancer chemotherapy. Hence, hGST P1-1 can be a promising target for cancer treatment. In this study, new hGST P1-1 inhibitors, 2-(4-substitutedphenyl/benzyl)-5-(4-trifluoromethylphenylsulphonamido) benzoxazole derivatives (Va-Vk) have been designed and synthesized. Surprisingly, in vitro hGST P1-1 enzyme inhibition studies demonstrated that all of the tested compounds except Vj had better activity than the reference drug EA and it is also correlated with the docking results. Additionally we compared the interactions with hGST P1-1 enzyme of newly synthesized compound Vh (bearing CF3 group) and previously synthesized compound 5f (bearing NO2 group). According to the docking results, compound Vh bound to the hGST P1-1 enzyme with a higher affinity compared to 5f. Therefore, we can consider that these data make a sense and can explain its higher activity. The compounds that obtained from this research could be used as scaffolds in design of new potent hGST P1-1 inhibitors useful in the treatment of the resistance of cancer chemotherapy.


Nondetection sampling bias in marked presence-only data.
Species distribution models (SDM) are tools used to determine environmental features that influence the geographic distribution of species' abundance and have been used to analyze presence-only records. Analysis of presence-only records may require correction for nondetection sampling bias to yield reliable conclusions. In addition, individuals of some species of animals may be highly aggregated and standard SDMs ignore environmental features that may influence aggregation behavior.We contend that nondetection sampling bias can be treated as missing data. Statistical theory and corrective methods are well developed for missing data, but have been ignored in the literature on SDMs. We developed a marked inhomogeneous Poisson point process model that accounted for nondetection and aggregation behavior in animals and tested our methods on simulated data.Correcting for nondetection sampling bias requires estimates of the probability of detection which must be obtained from auxiliary data, as presence-only data do not contain information about the detection mechanism. Weighted likelihood methods can be used to correct for nondetection if estimates of the probability of detection are available. We used an inhomogeneous Poisson point process model to model group abundance, a zero-truncated generalized linear model to model group size, and combined these two models to describe the distribution of abundance. Our methods performed well on simulated data when nondetection was accounted for and poorly when detection was ignored.We recommend researchers consider the effects of nondetection sampling bias when modeling species distributions using presence-only data. If information about the detection process is available, we recommend researchers explore the effects of nondetection and, when warranted, correct the bias using our methods. We developed our methods to analyze opportunistic presence-only records of whooping cranes (Grus americana), but expect that our methods will be useful to ecologists analyzing opportunistic presence-only records of other species of animals.


Design and fabrication of computer-generated beam-shaping holograms.
For the design of computer-generated holograms reconstructing certain intensity patterns with phase freedom, we use an object-oriented approach. The given intensity pattern is decomposed into elementary objects for which appropriate phase-only hologram functions can be constructed. The total hologram function is found by the subsequent superposition of its constituents, with a relative amplitude and phase weighting for each of them. Thus, the degrees of freedom are dramatically reduced compared with those of sampling approaches. The design algorithm allows us to compensate on the one hand for the intensity and phase distribution of the impinging laser beam and on the other hand for the shape of the hologram aperture. We report on the computer-aided design of such holograms, as well as their fabrication through the use of laser lithography and reactive ion etching. Optical reconstructions are shown.


Interferon regulatory factor 7 regulates glioma stem cells via interleukin-6 and Notch signalling.
Inflammatory microenvironment signalling plays a crucial role in tumour progression (i.e. cancer cell proliferation, survival, angiogenesis and metastasis) in many types of human malignancies. However, the role of inflammation in brain tumour pathology remains poorly understood. Here, we report that interferon regulatory factor 7 is a crucial regulator of brain tumour progression and heterogeneity. Ectopic expression of interferon regulatory factor 7 in glioma cells promotes tumorigenicity, angiogenesis, microglia recruitment and cancer stemness in vivo and in vitro through induction of interleukin 6, C-X-C motif chemokine 1 and C-C motif chemokine 2. In particular, interferon regulatory factor 7-driven interleukin 6 plays a pivotal role in maintaining glioma stem cell properties via Janus kinase/signal transducer and activator of transcription-mediated activation of Jagged-Notch signalling in glioma cells and glioma stem cells derived from glioma patients. Accordingly, the short hairpin RNA-mediated depletion of interferon regulatory factor 7 in glioma stem cells markedly suppressed interleukin 6-Janus kinase/signal transducer and activator of transcription-mediated Jagged-Notch-signalling pathway, leading to decreases in glioma stem cell marker expression, tumoursphere-forming ability, and tumorigenicity. Furthermore, in a mouse model of wound healing, depletion of interferon regulatory factor 7 suppressed tumour progression and decreased cellular heterogeneity. Finally, interferon regulatory factor 7 was overexpressed in patients with high-grade gliomas, suggesting its potential as an independent prognostic marker for glioma progression. Taken together, our findings indicate that interferon regulatory factor 7-mediated inflammatory signalling acts as a major driver of brain tumour progression and cellular heterogeneity via induction of glioma stem cell genesis and angiogenesis.


An essential role for UTX in resolution and activation of bivalent promoters.
Trimethylated histone H3 lysine 27 (H3K27me3) is linked to gene silencing, whereas H3K4me3 is associated with gene activation. These two marks frequently co-occupy gene promoters, forming bivalent domains. Bivalency signifies repressed but activatable states of gene expression and can be resolved to active, H3K4me3-prevalent states during multiple cellular processes, including differentiation, development and epithelial mesenchymal transition. However, the molecular mechanism underlying bivalency resolution remains largely unknown. Here, we show that the H3K27 demethylase UTX (also called KDM6A) is required for the resolution and activation of numerous retinoic acid (RA)-inducible bivalent genes during the RA-driven differentiation of mouse embryonic stem cells (ESCs). Notably, UTX loss in mouse ESCs inhibited the RA-driven bivalency resolution and activation of most developmentally critical homeobox (Hox) a-d genes. The UTX-mediated resolution and activation of many bivalent Hox genes during mouse ESC differentiation were recapitulated during RA-driven differentiation of human NT2/D1 embryonal carcinoma cells. In support of the importance of UTX in bivalency resolution, Utx-null mouse ESCs and UTX-depleted NT2/D1 cells displayed defects in RA-driven cellular differentiation. Our results define UTX as a bivalency-resolving histone modifier necessary for stem cell differentiation.


Psychometric properties of the Parenting Concerns Questionnaire in cancer survivors with minor and young adult children.
Although cancer patients with minor children have become more of a focus of psycho-oncological research, little is known about specific parenting concerns. Instruments to assess the concerns and worries of parents with cancer are rare. The Parenting Concerns Questionnaire (PCQ) addresses this issue. We analyzed parenting concerns in cancer survivors and evaluated the German version of the PCQ. A total of 1416 cancer survivors with minor and young adult children (≤21 years) were recruited in a register-based study. Descriptive analyses as well as reliability and validity analyses were conducted. We performed a confirmatory factorial analysis of the factor structure proposed by the authors of the original version on the PCQ. Seventy-three percent of the cancer survivors were women, average age was 47.5 years (SD 5.9). Mean time since diagnosis was 44 months (SD 23.4). Between 18 and 31% of survivors reported that they were concerned about their children. The PCQ proved to be a reliable and valid instrument showing medium correlations with standardized measures in expected directions and discriminating between survivors with and without use of psychosocial support services. The factor structure was supported by the confirmatory factorial analysis. Assessing parenting concerns gives an additional insight into the situation of parents with cancer. In our sample of cancer survivors, we identified one out of three survivors being concerned regarding the impact of their illness on their children. The PCQ can be considered as a valid and reliable instrument with regard to identifying concerned parents with cancer. Copyright © 2015 John Wiley & Sons, Ltd.


Modulation of cytosolic calcium signaling by protein kinase A-mediated phosphorylation of inositol 1,4,5-trisphosphate receptors.
Calcium release via intracellular Ca2+ release channels is a central event underpinning the generation of numerous, often divergent physiological processes. In electrically non-excitable cells, this Ca2+ release is brought about primarily through activation of inositol 1,4,5-trisphosphate receptors and typically takes the form of calcium oscillations. It is widely believed that information is carried in the temporal and spatial characteristics of these signals. Furthermore, stimulation of individual cells with different agonists can generate Ca2+ oscillations with dramatically different spatial and temporal characteristics. Thus, mechanisms must exist for the acute regulation of Ca2+ release such that agonist-specific Ca2+ signals can be generated. One such mechanism by which Ca2+ signals can be modulated is through simultaneous activation of multiple second messenger pathways. For example, activation of both the InsP3 and cAMP pathways leads to the modulation of Ca2+ release through protein kinase A mediated phosphoregulation of the InsP3R. Indeed, each InsP3R subtype is a potential substrate for PKA, although the functional consequences of this phosphorylation are not clear. This review will focus on recent advances in our understanding of phosphoregulation of InsP3R, as well as the functional consequences of this modulation in terms of eliciting specific cellular events.


Imaging the magnetic structures of artificial quasicrystal magnets using resonant coherent diffraction of circularly polarized X-rays.
Unraveling nanoscale spin structures has long been an important activity addressing various scientific interests, that are also readily adaptable to technological applications. This has invigorated the development of versatile nanoprobes suitable for imaging specimens under native conditions. Here we have demonstrated the resonant coherent diffraction of an artificial quasicrystal magnet with circularly polarized X-rays. The nanoscale magnetic structure was revealed from X-ray speckle patterns by comparing with micromagnetic simulations, as a step toward understanding the intricate relationship between the chemical and spin structures in an aperiodic quasicrystal lattice. Femtosecond X-ray pulses from free electron lasers are expected to immediately extend the current work to nanoscale structure investigations of ultrafast spin dynamics, surpassing the present spatio-temporal resolution.


The Effect of Physical Therapy Treatment in Patients with Subjective Tinnitus: A Systematic Review.
Background: Tinnitus is a very common symptom that often causes distress and decreases the patient's quality of life. Apart from the well-known causes, tinnitus can in some cases be elicited by dysfunctions of the cervical spine or the temporomandibular joint (TMJ). To date however, it is unclear whether alleviation of these dysfunctions, by physical therapy treatment, also decreases the tinnitus complaints. Such physical therapy could be an interesting treatment option for patients that are now often left without treatment. Objectives: The aim of this review was to investigate the current evidence regarding physical therapy treatment in patients with tinnitus. Data sources: The online databases Pubmed, Web of Science, Cochrane, and Embase were searched up to March 2016. Two independent reviewers conducted the data extraction and methodological quality assessment. Study eligibility criteria: Only randomized controlled trials and quasi-experimental trials were included in the review. Studies had to be written in English, French, Dutch, or German. Participants and interventions: The included studies investigated the effect of physical therapy treatment modalities on tinnitus severity in patients suffering from subjective tinnitus. Results: Six studies were included in this review, four investigating cervical spine treatment and two investigating TMJ treatment. These studies show positive effects of cervical spine treatment (manipulations, exercises, triggerpoint treatment) on tinnitus severity. Additionally, decrease in tinnitus severity and intensity was demonstrated after TMJ treatment, following splints, occlusal adjustments as well as jaw exercises. Limitations: The risk of bias in the included studies was high, mainly due to lack of randomization, lack of blinding of subjects, therapists, and/or investigators. Additionally, risk of bias is present due to incomplete presentation of the data and selective reporting. A major issue of the reviewed papers is the heterogeneity of the included study populations, treatments and outcome measures, which inhibit data pooling and meta-analysis. Conclusions: Despite the methodological issues in the included studies and the consequent low quality evidence, it is noteworthy that all included studies show positive treatment effects. Before recommendations can be made, these results need to be confirmed in larger, high quality studies, using unambiguous inclusion criteria, state-of-the-art treatment, and high quality outcome measures.


Estimating the association between SF-12 responses and EQ-5D utility values by response mapping.
Reliably mapping from generic or disease-specific health status measures into health state utilities would assist health economists. Existing studies mainly use ordinary least squares (OLS) regression equations to predict utility values for particular health states. The authors examine an alternative approach to map between 2 generic health status instruments, the SF-12 and the EQ-5D. Multinomial logit regression is used to estimate the probability that a respondent will select a particular level of response to questions in the EQ-5D, using individual question responses and summary scores from the SF-12 as predictors. Monte Carlo simulation methods are used to generate predicted EQ-5D responses, and utility scores (tariffs) are then attached. Results are compared with an alternative approach based on direct mapping to utility scores using OLS. The authors estimate equations using 12,967 adult survey responses-from the 2000 US Medical Expenditure Panel Survey. They report mean squared error (MSE) and mean absolute error (MAE) of their predicted utilities within this sample, and out-of-sample using 13,304 adults from the 1996 Health Survey for England. The authors obtain an in-sample and out-of-sample MSE of 0.03, compared with 0.02 for the OLS approach. Their MAE of 0.11 is similar to OLS results. The authors' method predicts group mean utility scores and differentiates between groups with or without known existing illness. The authors' approach has higher MSE than the direct OLS approach but gives more descriptive data on domains of health effects. Further out of sample prediction work will help test the validity of these methods.


Colors with functions: elucidating the biochemical and molecular basis of carotenoid metabolism.
Carotenoids affect a rich variety of physiological functions in nature and are beneficial for human health, serving as antioxidants in lipophilic environments and blue light filters in the macula of human retina. These dietary compounds also serve as precursors of a unique set of apo-carotenoid cleavage products, including retinoids. Although knowledge about retinoid biology has tremendously increased, the metabolism of retinoids' parent precursors remains poorly understood. Recently, molecular players in carotenoid metabolism have been identified and biochemically characterized. Moreover, mutations in their corresponding genes impair carotenoid metabolism and induce various pathologies in animal models. Polymorphisms in these genes alter carotenoid and retinoid homeostasis in humans as well. This review summarizes our current knowledge about the molecular/biochemical basis of carotenoid metabolism and particularly the physiological role of carotenoids in retinoid-dependent physiological processes.


Characterization of muscarinic cholinergic receptors in human and dog cerebral arteries.
Various concentrations of acetylcholine (ACh) produced dose dependent relaxations of isolated, helical preparations of human cerebral arteries and these responses were blocked by atropine. The median effective concentration (EC50) of ACh was 6.1 +/- 0.2 X 10(-6)M. ACh produced dual responses in isolated dog cerebral arteries. ACh in low concentrations produced relaxation, and contraction occurred when the concentration was raised to 1 X 10(-5)M. The EC50 of ACh which produced relaxation, in dog cerebral arteries was 7.2 +/- 0.2 X 10(-7)M. Muscarinic cholinergic receptors in these arteries were analyzed directly using 3H-QNB as the ligand. The specific 3H-QNB binding to the arteries was saturable and of KD = 1.5 nM and Bmax = 93 fmol/mg protein in human cerebral arteries and KD = 0.59 nM, Bmax = 340 fmol/mg protein in dog cerebral arteries. Specific binding of 3H-QNB was displaced by muscarinic cholinergic agents. Ki values and Hill coefficients were as follows: QNB, 1.0 X 10(-9)M, 0.89; atropine, 1.1 X 10(-8)M, 0.95; ACh, 0.8 X 10(-8)M and 2.1 X 10(-6)M, 0.54; carbachol, 1.2 X 10(-7)M and 4.3 X 10(-5)M, 0.33 in human cerebral arteries and QNB, 0.55 X 10(-9)M, 0.85; atropine 0.9 X 10(-9)M, 1.00; ACh, 0.9 X 10(-9)M and 1.1 X 10(-6)M, 0.43; carbachol 6.3 X 10(-8)M and 1.1 X 10(-5)M, 0.36 in dog cerebral arteries. Endogenous choline acetyltransferase (ChAT) activity was 1.6 +/- 0.4 nmol/mg protein/hr in human cerebral arteries and 3.7 +/- 0.1 nmol/mg protein/hr in dog cerebral arteries.(ABSTRACT TRUNCATED AT 250 WORDS)