Impact of primer dimers and self-amplifying hairpins on reverse transcription loop-mediated isothermal amplification detection of viral RNA.
Loop-mediated isothermal amplification (LAMP), coupled with reverse transcription (RT), has become a popular technique for detection of viral RNA due to several desirable characteristics for use in point-of-care or low-resource settings. The large number of primers in LAMP (six per target) leads to an increased likelihood of primer dimer interactions, and the inner primers in particular are prone to formation of stable hairpin structures due to their length (typically 40-45 bases). Although primer dimers and hairpin structures are known features to avoid in nucleic acid amplification techniques, there is little quantitative information in literature regarding the impact of these structures on LAMP or RT-LAMP assays. In this study, we examine the impact of primer dimers and hairpins on previously published primer sets for dengue virus and yellow fever virus. We demonstrate that minor changes to the primers to eliminate amplifiable primer dimers and hairpins improves the performance of the assays when monitored in real time with intercalating dyes, and when monitoring a fluorescent endpoint using the QUASR technique. We also discuss the thermodynamic implications of these minor changes on the overall stability of amplifiable secondary structures, and we present a single thermodynamic parameter that can be correlated to the probability of non-specific amplification associated with LAMP primers.


Touch Perception Altered by Chronic Pain and by Opioid Blockade.
Touch plays a significant role in human social behavior and social communication, and its rewarding nature has been suggested to involve opioids. Opioid blockade in monkeys leads to increased solicitation and receipt of grooming, suggesting heightened enjoyment of touch. We sought to study the role of endogenous opioids in perception of affective touch in healthy adults and in patients with fibromyalgia, a chronic pain condition shown to involve reduced opioid receptor availability. The pleasantness of touch has been linked to the activation of C-tactile fibers, which respond maximally to slow gentle touch and correlate with ratings of pleasantness. We administered naloxone to patients and healthy controls to directly observe the consequences of µ-opioid blockade on the perceived pleasantness and intensity of touch. We found that at baseline chronic pain patients showed a blunted distinction between slow and fast brushing for both intensity and pleasantness, suggesting reduced C-tactile touch processing. In addition, we found a differential effect of opioid blockade on touch perception in healthy subjects and pain patients. In healthy individuals, opioid blockade showed a trend toward increased ratings of touch pleasantness, while in chronic pain patients it significantly decreased ratings of touch intensity. Further, in healthy individuals, naloxone-induced increase in touch pleasantness was associated with naloxone-induced decreased preference for slow touch, suggesting a possible effect of opioid levels on processing of C-tactile fiber input. These findings suggest a role for endogenous opioids in touch processing, and provide further evidence for altered opioid functioning in chronic pain patients.


Effects of the selective MPS1 inhibitor MPS1-IN-3 on glioblastoma sensitivity to antimitotic drugs.
Glioblastomas exhibit a high level of chemotherapeutic resistance, including to the antimitotic agents vincristine and taxol. During the mitotic agent-induced arrest, glioblastoma cells are able to perform damage-control and self-repair to continue proliferation. Monopolar spindle 1 (MPS1/TTK) is a checkpoint kinase and a gatekeeper of the mitotic arrest. We used glioblastoma cells to determine the expression of MPS1 and to determine the effects of MPS1 inhibition on mitotic errors and cell viability in combination with vincristine and taxol. The effect of MPS1 inhibition was assessed in different orthotopic glioblastoma mouse models (n = 3-7 mice/group). MPS1 expression levels were examined in relation to patient survival. Using publicly available gene expression data, we determined that MPS1 overexpression corresponds positively with tumor grade and negatively with patient survival (two-sided t test, P < .001). Patients with high MPS1 expression (n = 203) had a median and mean survival of 487 and 913 days (95% confidence intervals [CI] = 751 to 1075), respectively, and a 2-year survival rate of 35%, whereas patients with intermediate MPS1 expression (n = 140) had a median and mean survival of 858 and 1183 days (95% CI = 1177 to 1189), respectively, and a 2-year survival rate of 56%. We demonstrate that MPS1 inhibition by RNAi results in sensitization to antimitotic agents. We developed a selective small-molecule inhibitor of MPS1, MPS1-IN-3, which caused mitotic aberrancies in glioblastoma cells and, in combination with vincristine, induced mitotic checkpoint override, increased aneuploidy, and augmented cell death. MPS1-IN-3 sensitizes glioblastoma cells to vincristine in orthotopic mouse models (two-sided log-rank test, P < .01), resulting in prolonged survival without toxicity. Our results collectively demonstrate that MPS1, a putative therapeutic target in glioblastoma, can be selectively inhibited by MPS1-IN-3 sensitizing glioblastoma cells to antimitotic drugs.


Protein signatures of centenarians and their offspring suggest centenarians age slower than other humans.
Using samples from the New England Centenarian Study (NECS), we sought to characterize the serum proteome of 77 centenarians, 82 centenarians' offspring, and 65 age-matched controls of the offspring (mean ages: 105, 80, and 79 years). We identified 1312 proteins that significantly differ between centenarians and their offspring and controls (FDR < 1%), and two different protein signatures that predict longer survival in centenarians and in younger people. By comparing the centenarian signature with 2 independent proteomic studies of aging, we replicated the association of 484 proteins of aging and we identified two serum protein signatures that are specific of extreme old age. The data suggest that centenarians acquire similar aging signatures as seen in younger cohorts that have short survival periods, suggesting that they do not escape normal aging markers, but rather acquire them much later than usual. For example, centenarian signatures are significantly enriched for senescence-associated secretory phenotypes, consistent with those seen with younger aged individuals, and from this finding, we provide a new list of serum proteins that can be used to measure cellular senescence. Protein co-expression network analysis suggests that a small number of biological drivers may regulate aging and extreme longevity, and that changes in gene regulation may be important to reach extreme old age. This centenarian study thus provides additional signatures that can be used to measure aging and provides specific circulating biomarkers of healthy aging and longevity, suggesting potential mechanisms that could help prolong health and support longevity.


StratomeX: Visual Analysis of Large-Scale Heterogeneous Genomics Data for Cancer Subtype Characterization.
Identification and characterization of cancer subtypes are important areas of research that are based on the integrated analysis of multiple heterogeneous genomics datasets. Since there are no tools supporting this process, much of this work is done using ad-hoc scripts and static plots, which is inefficient and limits visual exploration of the data. To address this, we have developed StratomeX, an integrative visualization tool that allows investigators to explore the relationships of candidate subtypes across multiple genomic data types such as gene expression, DNA methylation, or copy number data. StratomeX represents datasets as columns and subtypes as bricks in these columns. Ribbons between the columns connect bricks to show subtype relationships across datasets. Drill-down features enable detailed exploration. StratomeX provides insights into the functional and clinical implications of candidate subtypes by employing small multiples, which allow investigators to assess the effect of subtypes on molecular pathways or outcomes such as patient survival. As the configuration of viewing parameters in such a multi-dataset, multi-view scenario is complex, we propose a meta visualization and configuration interface for dataset dependencies and data-view relationships. StratomeX is developed in close collaboration with domain experts. We describe case studies that illustrate how investigators used the tool to explore subtypes in large datasets and demonstrate how they efficiently replicated findings from the literature and gained new insights into the data.


Dynamic aspects of guinea pig inner hair cell receptor potentials with transient asphyxia.
DC and AC receptor potentials of cochlear inner hair cells in response to tone bursts of various frequencies and intensities were continuously measured during and following periods of transient asphyxia. The effects of asphyxia were most pronounced for low sound pressure level (SPL) acoustic stimuli near the characteristic frequency (CF) of the inner hair cell, leading to vulnerability of the 'tip' of the cell's frequency tuning curve (FTC). The resulting changes in the shape of the FTC are, first, a reduction in tip criterion sensitivity of 10-20 dB without significant loss in sharpness of tuning. Later, when the full effect of 30-45 s asphyxia occurs, tip sensitivity loss between 30 and 65 dB is accompanied by greatly broadened tuning and a shift downward in frequency of the CF by greater than 1/4 octave. The CF shift is due to a progressive loss of high frequency sensitivity. The linear segment of the input-output (intensity) function, plotted as log DC receptor potential versus SPL (at the original CF), becomes longer during the early phase asphyxia, and the slope of the segment declines by 50%. At high SPLs, for all frequencies, the time course of the receptor potential change was similar in shape to that exhibited by the endocochlear potential (EP). In particular, for high sound levels, the recovery of response matches the EP while for low level tip frequency sounds recovery is protracted. No difference between the decline of the AC and DC receptor potentials at CF was observed. Inner hair cell resting membrane potential (Em) hyperpolarized during asphyxia by 2-6 mV, correlating with the change in EP according to a ratio of 1/10 (Em/EP).


The putative role of the intestinal microbiota in the irritable bowel syndrome.
The irritable bowel syndrome (IBS) is a chronic abdominal symptom complex that is heterogeneous in terms of its clinical presentation and underlying pathophysiology and pathogenesis. It is now established that enteric infection can trigger the syndrome in at least a subset of patients. In addition, there is growing evidence of low grade inflammation and immune activation in the distal bowel of some IBS patients. These observations now prompt the question as to what maintains gut dysfunction in these patients. The intestinal microbiota influences a broad array of host organs that include the gut and the brain, and is an important determinant of normal function in these systems. Disruption of the delicate balance between the host and its intestinal microbiota (termed dysbiosis) results in changes in the mucosal immune system that range from overt inflammation as seen in Crohn's Disease, to low grade inflammation without tissue injury, as seen in a subset of IBS patients. Under experimental conditions, disruption of the microbiota also produces changes in gut sensory-motor function and immune activity. Thus, dysbiosis induced by infection, dietary change or drugs such as antibiotics could produce low grade inflammation and chronic gut dysfunction, reminiscent of that seen in IBS. Fluctuations in gut physiology destabilize the habitat of commensal bacteria and provide a basis for chronic dysbiosis. Recent observations in animal models that changes in gut flora influence behavior provide a basis for a novel unifying hypothesis that accommodates both gut dysfunction and behavioral changes that characterize many IBS patients. This hypothesis states that dysbiosis exists in at least a subset of IBS patients, as a result of infection, dietary change or drugs and contributes to gut inflammatory and functional change in addition to psychiatric co-morbidity.


Drosophila nuclear proteins bind to regions of alternating C and T residues in gene promoters.
Proteins from Drosophila nuclei that bind to regions of alternating C and T residues present in the promoters of the heat shock genes hsp70 and hsp26 and the histone genes his3 and his4 have been purified. These proteins bind to isolated linear DNA, and genomic footprinting analyses indicate that they are bound to DNA in nuclei. In supercoiled plasmids at low pH, some of these DNA sequences adopt triple-helical structures which, if they form in vivo, could significantly affect chromatin structure. The nuclear proteins described here, and not necessarily the deformed conformation of the DNA, may be responsible for maintaining a potentially inducible promoter structure before transcriptional activation.


Outcomes of extubation failure in medical intensive care unit patients.
Extubation failure is associated with a poor prognosis, but the respective roles for reintubation per se and underlying disease severity remain unclear. Our objectives were to evaluate the impact of failed extubation, whether planned or unplanned, on patient outcomes and to identify a patient subset at risk for extubation failure. Prospective 1-yr observational study with daily data collection. : Thirteen-bed medical intensive care unit in a teaching hospital. Consecutive patients requiring invasive mechanical ventilation were screened and followed until discharge or death. None. Of 168 planned extubations in 340 patients, 26 (15%) failed. Of these 26 patients, seven (27%) had pneumonia and 13 (50%) died after reintubation. Compared with successfully extubated patients, the patients with failed extubation were not significantly different regarding disease severity, mechanical ventilation duration, or blood gas values. Age and underlying diseases were the only factors associated with extubation failure, and extubation failure occurred in 34% of patients >65 yrs with chronic cardiac or respiratory disease compared with only 9% of other patients (p < .01). Unplanned extubation occurred in 9% of patients, and inadequate endotracheal tube position was a risk factor. Failure of both planned and unplanned extubation was specifically associated with significant rapid worsening of daily organ dysfunction scores. Patients >65 yrs with underlying chronic cardiac or respiratory disease are at high risk for extubation failure and subsequent pneumonia and death. Contrasting with successful extubation, failed planned or unplanned extubation was followed by marked clinical deterioration, suggesting a direct and specific effect of extubation failure and reintubation on patient outcomes.


Transcriptional Modulator Ifrd1 Regulates Osteoclast Differentiation through Enhancing the NF-κB/NFATc1 Pathway.
Bone homeostasis is maintained by the synergistic actions of bone-resorbing osteoclasts and bone-forming osteoblasts. Here, we show that the transcriptional coactivator/repressor interferon-related developmental regulator 1 (Ifrd1) is expressed in osteoclast lineages and represents a component of the machinery that regulates bone homeostasis. Ifrd1 expression was transcriptionally regulated in preosteoclasts by receptor activator of nuclear factor κB (NF-κB) ligand (RANKL) through activator protein 1. Global deletion of murine Ifrd1 increased bone formation and decreased bone resorption, leading to a higher bone mass. Deletion of Ifrd1 in osteoclast precursors prevented RANKL-induced bone loss, although no bone loss was observed under normal physiological conditions. RANKL-dependent osteoclastogenesis was impaired in vitro in Ifrd1-deleted bone marrow macrophages (BMMs). Ifrd1 deficiency increased the acetylation of p65 at residues K122 and K123 via the inhibition of histone deacetylase-dependent deacetylation in BMMs. This repressed the NF-κB-dependent transcription of nuclear factor of activated T cells, cytoplasmic 1 (NFATc1), an essential regulator of osteoclastogenesis. These findings suggest that an Ifrd1/NF-κB/NFATc1 axis plays a pivotal role in bone remodeling in vivo and represents a therapeutic target for bone diseases.


Respective clustering of unfavorable and favorable cytogenetic clones in myelofibrosis with myeloid metaplasia with homozygosity for JAK2(V617F) and response to erythropoietin therapy.
Patients who have myelofibrosis with myeloid metaplasia (MMM) display recurrent, albeit nonspecific cytogenetic abnormalities that are diverse prognostically. For the current study, the authors explored the relation between specific cytogenetic clones and JAK2(V617F) mutational status in patients with MMM and the effects on treatment response to erythropoietin (Epo). Concomitantly collected blood granulocytes and bone marrow were processed for JAK2(V617F) mutation analysis and cytogenetic studies, respectively. Genomic DNA was amplified by polymerase chain reaction, and fluorescent dye chemistry sequencing was performed by using the same primers that were used for amplification. Among 105 study patients, cytogenetic abnormalities were detected in 47 patients (45%), and the JAK2(V617F) mutation was detected in 52 patients (50%). Comparison of mutational frequencies between favorable (normal, sole 13q-, or 20q- clones; n = 70 patients) and unfavorable (all other abnormalities; n = 35 patients) cytogenetic categories revealed a significantly different incidence of homozygous JAK2(V617F) between them (9% vs. 23%, respectively; P = .04). Furthermore, the mutant allele coexisted with several recurrent cytogenetic lesions. Among 25 patients who received Epo either alone (n = 17 patients) or in combination with hydroxyurea (n = 8 patients), 4 patients (16%) achieved a response, and none of them were homozygous for JAK2(V617F). Conversely, a response was more likely (P = .0001) in the presence of favorable cytogenetic abnormalities (i.e., 3 of 4 responders carried sole 13q- or 20q- clones). Unfavorable and favorable cytogenetic clones in MMM clustered with homozygosity for JAK2(V617F) and treatment response to Epo-based therapy, respectively.


Selective inhibition of the accumulation of extracellular proteases of Pseudomonas aeruginosa by gentamicin and tobramycin.
Gentamicin and tobramycin inhibited the accumulation of extracellular proteases secreted by Pseudomonas aeruginosa. The secretion of protease was inhibited at concentrations of these drugs that were below the level required to inhibit general protein synthesis. Neither magnesium ions nor high ionic strength antagonized the ability of the aminoglycosides to block secretion of the proteases. Under these culture conditions magnesium ions were shown to antagonize the effects of the aminoglycosides on protein synthesis and aminoglycoside-mediated lysozyme lysis of P. aeruginosa. These results suggested that the drugs blocked secretion of the proteases by acting at the level of the outer membrane.


Solid-state NMR and SAXS studies provide a structural basis for the activation of alphaB-crystallin oligomers.
The small heat shock protein alphaB-crystallin (alphaB) contributes to cellular protection against stress. For decades, high-resolution structural studies on oligomeric alphaB have been confounded by its polydisperse nature. Here, we present a structural basis of oligomer assembly and activation of the chaperone using solid-state NMR and small-angle X-ray scattering (SAXS). The basic building block is a curved dimer, with an angle of approximately 121 degrees between the planes of the beta-sandwich formed by alpha-crystallin domains. The highly conserved IXI motif covers a substrate binding site at pH 7.5. We observe a pH-dependent modulation of the interaction of the IXI motif with beta4 and beta8, consistent with a pH-dependent regulation of the chaperone function. N-terminal region residues Ser59-Trp60-Phe61 are involved in intermolecular interaction with beta3. Intermolecular restraints from NMR and volumetric restraints from SAXS were combined to calculate a model of a 24-subunit alphaB oligomer with tetrahedral symmetry.


The relationship between disordered eating and cigarette smoking among adult female twins.
While previous studies have shown a link between cigarette smoking and disordered eating (DE), it is less clear whether this association can be explained by attempts to control weight and/or temperament. This study had two aims: to examine weight/shape and DE symptomatology associated with weight control-related smoking and disordered eating; second, to investigate whether temperament-based factors explain the association between compensatory smoking and weight and shape-related characteristics, and disordered eating symptoms. Female twins (N = 1,002) from the Australian Twin Registry, aged 28-39 years, were assessed by interview yielding lifetime diagnostic information of disordered eating and use of cigarette smoking for weight control. Self-report measures of temperament were available. Women who had never smoked (50.6%, n = 495) or had smoked for reasons other than weight and shape control (42.5%, n = 415) were generally not differentiated with respect to indicators of disordered eating. Women who smoked for reasons primarily associated with weight and shape control (6.9%, n = 67) had significantly higher levels of disordered eating than non-smokers identified as being higher levels of purging (OR = 4.55, 95% CI = 2.41-8.59) and fasting (OR = 9.32, 94% CI = 4.43-19.90) but not objective binge episodes (OR = 1.69, 95% CI = 0.71-3.99). Of those women diagnosed with lifetime eating disorders, weight-control smokers had higher levels of purging (OR = 4.22, 95% CI = 1.13-15.80) than those who did not. There were no differences in temperament. Results support co-occurrence of smoking for weight control and purging in both community and clinical groups, and indicate this association is related to weight control and not temperament.


The effects of two nights of sleep deprivation with or without energy restriction on immune indices at rest and in response to cold exposure.
The purpose of the study was to determine the effects of two nights of sleep deprivation with or without energy restriction on immune indices at rest and in response to cold exposure. On three randomised occasions ten males slept normally [mean (SD): 436 (21) min night(-1); CON], were totally sleep-deprived (SDEP), or were totally sleep-deprived and 90% energy-restricted (SDEP + ER) for 53 h. After 53 h (1200 h) participants performed a seated cold air test (CAT) at 0.0 degrees C until T (re) decreased to 36.0 degrees C. Circulating leucocyte counts, neutrophil degranulation, stress hormones and saliva secretory IgA (S-IgA) were determined at 0 h, 24 h, 48 h, pre-CAT, post-CAT, 1-h and 2-h post-CAT. One night on SDEP increased bacterially stimulated neutrophil degranulation (21%, P < 0.05), and two nights on SDEP and SDEP + ER increased S-IgA concentration (40 and 44%; P < 0.01). No other significant effects were observed for immuno-endocrine measures prior to CAT. CAT duration was not different between trials [mean (SD): 133 (53) min] and T (re) decreased to 35.9 (0.3) degrees C. Modest whole-body cooling decreased circulating lymphocyte counts (25%; P < 0.01), S-IgA concentration (36%; P < 0.01) and secretion rate (24%; P < 0.05). A neutrophilia occurred post-CAT on CON and SDEP and 2-h post-CAT on SDEP + ER (P < 0.01). Modest whole-body cooling also decreased neutrophil degranulation on CON (22%) and SDEP (18%; P < 0.05). Plasma cortisol and norepinephrine increased post-CAT (31 and 346%, P < 0.05), but modest whole-body cooling did not alter plasma epinephrine. In conclusion, two nights of SDEP or SDEP + ER did not compromise resting immune indices. However, modest whole-body cooling (T(re) 35.9 degrees C) decreased circulating lymphocytes, neutrophil degranulation and S-IgA, but responses were not amplified by prior SDEP or SDEP + ER.


Copurification of small heat shock protein with alpha B crystallin from human skeletal muscle.
Immunoreactive alpha B crystallin and a 28-kDa protein in an extract of human pectoral muscle were precipitated by (NH4)2SO4 at 40% saturation, and coeluted during column chromatography on DEAE-Sepharose and on Bio-Gel A-5m. The two proteins were separated on a column of S-Sepharose HP in the presence of 7 M urea. Further chromatography of each of the two resultant fractions on a column of Superdex 75 pg and on a TSK-SP 5PW column in the presence of urea yielded preparations of alpha B crystallin and the 28-kDa protein each of which gave a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The final preparation of 28-kDa protein contained at least two subtypes, which were separable on the TSK-SP column. However, fragmentation patterns of the two major 28-kDa proteins after digestion with endoproteinase Asp-N were identical. Amino acid sequences of peptides formed by cleavage of the purified 28-kDa protein and alpha B crystallin were identical to those of particular regions of the deduced amino acid sequences of human small heat shock protein (HSP28) and lens alpha B crystallin, respectively. Using an immunoassay method, with antibodies raised in rabbits, we found that HSP28 was present in all human tissues tested and at high levels (greater than 1 micrograms/mg protein) in the heart and other tissues composed of striated and smooth muscles. HSP28, found with alpha B crystallin, in extracts of several human and bovine tissues was trapped on and coeluted with alpha B crystallin from an affinity column prepared with antibodies against alpha B crystallin. This result suggests that the two proteins are associated in cells.


Comparative effects of H2-receptor antagonists on drug interaction in rats.
The most widely used H2-receptor antagonist, cimetidine, is known to interact with cytochrome P-450 drug-metabolizing enzymes and, therefore, interacts with other drugs which may be administered concurrently. In this study, effects of three H2-receptor antagonists, famotidine, ranitidine, and L-643,441, on drug interaction were studied using cimetidine as a positive control. Cimetidine and L-643,441, but not famotidine or ranitidine, prolonged antipyrine elimination and hexobarbital-induced sleeping time. The effect of cimetidine and famotidine on the anticoagulant effect on warfarin in rats was also investigated. Pretreatment of rats with cimetidine produced a significant depression of plasma prothrombin complex activity, whereas concomitant administration of famotidine did not alter the plasma prothrombin complex activity. Whereas cimetidine is known to impair the elimination of a number of drugs metabolized by microsomal mixed function oxidase enzyme systems, the results of the present study suggest that famotidine and ranitidine have little effect on these enzyme systems.


Immediate effects of kinesio taping on fixed postural alignment and foot balance in stroke patients.
[Purpose] The main aim of this study was to identify the short-term effects of Kinesio taping (KT) on the static body alignment and overall balance function presented by the coordinate and foot balance in stroke patients. [Subjects and Methods] Thirty-eight stroke subjects were randomly allocated into the study groups. The kinematic analysis measured deviation or changes from standard body alignment and foot pressure by the human anatomy-based coordinates were examined using the Shisei Innovation System PA200 ver.9.0. [Results] The glabellas on the front view, larynx on the front view, rt. greater tubercle of the humerus (vertical changes), lt. greater tubercle of the humerus (vertical changes), posterior superior iliac spine, and greater trochanter (horizontal changes) showed statistically significant decreases, indicating dislocation from the axis center, after taping. [Conclusion] The clinical use of KT for stroke patients who have asymmetrical and imbalanced body posture could be an optimal therapeutic approach. Since more evidence based practices are needed, future studies should include large numbers of subjects and examine diverse KT application patterns.


Intranasal dexmedetomidine premedication is comparable with midazolam in burn children undergoing reconstructive surgery.
Preoperative anxiety and emergence delirium in children continue to be common even with midazolam premedication. Midazolam is unpleasant tasting even with a flavored vehicle and as a result, patient acceptance is sometimes poor. As an alternative, we evaluated dexmedetomidine administered intranasally. Dexmedetomidine an alpha-2 adrenergic agonist is tasteless, odorless, and painless when administered by this route. Alpha-2 adrenergic agonists produce sedation, facilitate parental separation, and improve conditions for induction of general anesthesia, while preserving airway reflexes. Institutional review board approval was obtained to study 100 pediatric patients randomized to intranasal dexmedetomidine (2 microg/kg) or oral midazolam (0.5 mg/kg) administered 30 to 45 minutes before the surgery. Subjects received general anesthesia with oxygen, nitrous oxide, isoflurane, and analgesics (0.05-0.1 mg/kg morphine or 0.1 mg/kg methadone). Nurses and anesthetists were blinded to the drug administered and evaluated patients for preoperative sedation, conditions for induction of general anesthesia, emergence from anesthesia, and postoperative pain. Responses of 100 patients (50 dexmedetomidine and 50 midazolam) were analyzed. Dexmedetomidine (P=.003) was more effective than midazolam at inducing sleep preoperatively. Dexmedetomidine and midazolam were comparable for conditions at induction (P>0.05), emergence from anesthesia (P>0.05), or postoperative pain (P>0.05). Both drugs were equieffective in these regards. In pediatric patients, dexmedetomidine 2 microg/kg administered intranasally and midazolam 0.5 mg/kg administered orally produced similar conditions during induction and emergence of anesthesia. Intranasal administration of dexmedetomidine is more effective at inducing sleep and in some circumstances offers a useful alternative to oral midazolam in children.


Engaging Small Residential Construction Contractors in Community-Based Participatory Research to Promote Safety.
Construction is a large employment sector with a high prevalence of small businesses. Despite the high injury rates reported for employees of small construction firms, these firms are under-represented in occupational safety research studies. Such studies are needed to understand barriers experienced by these firms and to examine ways to overcome them. However, challenges accessing and recruiting this hard-to-reach population are frequently reported. Traditional approaches of recruiting through unions or workers' compensation insurers may not be appropriate or effective for small construction businesses. Previous studies have demonstrated the value of academic collaborations with community-based organizations for recruiting participants from hard-to-reach populations for research studies. In accordance with the principles of Community-Based Participatory Research (CBPR), we formed a recruitment team comprised of staff from a local union, a community organization, and a community outreach team to recruit small construction contractors in Lawrence, MA. Media marketing strategies, participation in community events, exploring neighborhoods in search of ongoing residential projects, and partnership with vocational training institutions and building trade associations were some of the strategies implemented during this project. We recruited 118 contractors, supervisors, and foremen from more than 50 construction firms across the Greater Lawrence area to participate in an intervention project to reduce falls and silica exposure. The CBPR approach facilitated the development and implementation of recruitment strategies that resulted in the participation of a significant number of hard-to-reach small construction contractors.


Embryonic temperature influences juvenile temperature choice and growth rate in snapping turtles Chelydra serpentina.
Snapping turtles (Chelydra serpentina) demonstrate temperature-dependent sex determination (TSD): intermediate egg incubation temperatures (23-27 degrees C) produce males, while extreme temperatures produce females. Snapping turtles are also sexually dimorphic: adult males are typically larger than females. Previous researchers hypothesized that male-producing egg temperatures enhanced the growth rate of juvenile turtles, resulting in the adult dimorphism and potentially providing an adaptive benefit for TSD. In reptiles, the choice of ambient temperature can also influence growth. I measured the effect of egg incubation temperature on juvenile growth rate and water temperature choice of C. serpentina. Eggs were incubated in the laboratory at 21.5, 24.5, 27.5 or 30.5 degrees C to produce both sexes, all males, both sexes or all females, respectively. Egg temperature was linearly and negatively correlated with growth rate of both male and female juveniles. Thus, growth was enhanced, but not maximized, by male-producing egg temperatures. Egg temperature was also negatively correlated with juvenile temperature choice such that, on average, turtles from 21.5 degrees C eggs selected 28 degrees C water, while turtles from 30.5 degrees C eggs chose 24.5 degrees C water. Additionally, these temperature choices were highly repeatable, even following a 6 month hibernation period at 7 degrees C. Thus, while male egg temperatures do not directly maximize growth, multiple effects of embryonic temperature may combine to create long-lasting differences in the behavioral physiology of male and female C. serpentina. Such differences could be important to the ecology and evolution of TSD.


Azathioprine in the treatment of autoimmune blistering diseases.
Although there are no standard guidelines for the treatment of autoimmune blistering diseases, azathioprine has shown good efficacy in acquired autoimmune blistering diseases, and is well tolerated. Side effects of azathioprine normally occur in mild variants. Severe reactions are due to reduced thiopurine S-methyltransferase (TPMT) or inosine triphosphate pyrophosphohydrolase (ITPA) activity. Therefore, screening for TPMT activity should be conducted in white patients and Africans, whereas Japanese should be screened for ITPA activity before therapy with azathioprine is started. Azathioprine is clinically meaningful for the treatment of pemphigus.


Low intravascular pressure activates endothelial cell TRPV4 channels, local Ca2+ events, and IKCa channels, reducing arteriolar tone.
Endothelial cell (EC) Ca(2+)-activated K channels (SK(Ca) and IK(Ca) channels) generate hyperpolarization that passes to the adjacent smooth muscle cells causing vasodilation. IK(Ca) channels focused within EC projections toward the smooth muscle cells are activated by spontaneous Ca(2+) events (Ca(2+) puffs/pulsars). We now show that transient receptor potential, vanilloid 4 channels (TRPV4 channels) also cluster within this microdomain and are selectively activated at low intravascular pressure. In arterioles pressurized to 80 mmHg, ECs generated low-frequency (~2 min(-1)) inositol 1,4,5-trisphosphate receptor-based Ca(2+) events. Decreasing intraluminal pressure below 50 mmHg increased the frequency of EC Ca(2+) events twofold to threefold, an effect blocked with the TRPV4 antagonist RN1734. These discrete events represent both TRPV4-sparklet- and nonsparklet-evoked Ca(2+) increases, which on occasion led to intracellular Ca(2+) waves. The concurrent vasodilation associated with increases in Ca(2+) event frequency was inhibited, and basal myogenic tone was increased, by either RN1734 or TRAM-34 (IK(Ca) channel blocker), but not by apamin (SK(Ca) channel blocker). These data show that intraluminal pressure influences an endothelial microdomain inversely to alter Ca(2+) event frequency; at low pressures the consequence is activation of EC IK(Ca) channels and vasodilation, reducing the myogenic tone that underpins tissue blood-flow autoregulation.


Obstructive urolithiasis in a Saanen goat in New Zealand, resulting in a ruptured bladder.
A 3-year-old male, castrated Saanen goat was presented with an acute onset of vocalisation, abdominal discomfort and 'tail-flagging'. Radiographs, including a positive-contrast study, indicated obstructive urolithiasis, and evidence of rupture of the urethra. Analysis of uroliths identified calcium carbonate crystals. Amputation of the urethral process could not re-establish urethral patency, so a tube cystotomy was performed but also was not successful. Eighteen days after admission, a perineal urethrostomy was performed. However, the goat's condition deteriorated 12 days after the urethrostomy, and the owner consented to euthanasia of the animal. Post-mortem examination revealed a hole 2x1 cm on the dorsal aspect of the bladder, and a necrotic focus measuring approximately 2x3 cm ventrally. Obstructive urolithiasis, with ruptured bladder and urethra. Obstructive urolithiasis is probably an under-reported condition of castrated male goats in New Zealand.


Invasion of mouse erythrocytes by the human malaria parasite, Plasmodium falciparum.
Plasmodium falciparum malaria merozoites require erythrocyte sialic acid for optimal invasion of human erythrocytes. Since mouse erythrocytes have the form of sialic acid found on human erythrocytes (N-acetyl neuraminic acid), mouse erythrocytes were tested for invasion in vitro. The Camp and 7G8 strains of P. falciparum invaded mouse erythrocytes at 17-45% of the invasion rate of human erythrocytes. Newly invaded mouse erythrocytes morphologically resembled parasitized human erythrocytes as shown on Giemsa-stained blood films and by electron microscopy. The rim of parasitized mouse erythrocytes contained the P. falciparum 155-kD protein, which is on the rim of ring-infected human erythrocytes. Camp but not 7G8 invaded rat erythrocytes, indicating receptor heterogeneity. These data suggest that it may be possible to adapt the asexual erythrocytic stage of P. falciparum to rodents. The development of a rodent model of P. falciparum malaria could facilitate vaccine development.


Poor oral hygiene and risk of esophageal squamous cell carcinoma in Kashmir.
Several studies have suggested an association between poor oral health and esophageal squamous cell carcinoma (ESCC). We conducted a case-control study in Kashmir, a region with relatively high incidence of ESCC in north India, to investigate the association between oral hygiene and ESCC risk. We recruited 703 histologically confirmed ESCC cases, and 1664 controls individually matched to the cases for age, sex, and district of residence. Conditional logistic regression models were used to calculate odds ratios (ORs) and 95% confidence intervals (CIs). We found an inverse association between teeth cleaning and ESCC risk. As compared with never cleaning teeth, the OR (95% CI) was 0.41 (0.28-0.62) for cleaning less than daily and 0.44 (0.25-0.77) for cleaning at least once a day (P for trend=0.026) in models adjusted for multiple potential confounders, including several indicators of socioeconomic status. This association persisted after we limited our analyses to never tobacco users. The inverse association between cleaning teeth and ESCC was stronger with using brushes than with using sticks/fingers. We also found an association between the number of decayed, filled, and missing teeth and ESCC risk, but the trend of the associations was not statistically significant. Avoiding solid food and cold beverages because of teeth and oral problems were also associated with ESCC risk. We found an association between poor oral hygiene indicators and ESCC risk, supporting the previous studies that showed the same associations.


Effects of genetic variants of human P450 oxidoreductase on catalysis by CYP2D6 in vitro.
Cytochrome P450 (P450) oxidoreductase (POR) donates electrons to all microsomal cytochrome P450s, including drug-metabolizing and steroidogenic enzymes. Severe POR mutations cause skeletal malformations and disordered steroidogenesis. The POR polymorphism A503V is found on approximately 28% of human alleles and decreases activities of CYP3A4 and steroidogenic CYP17, but not the activities of steroidogenic CYP21 or drug-metabolizing CYP1A2 and CYP2C19. CYP2D6 metabolizes about 25% of clinically used drugs; we assessed the capacity of POR variants to support the activities of human CYP2D6. N-27 forms of wildtype (WT), Q153R, A287P, R457H and A503V POR, and WT CYP2D6 were expressed in Escherichia coli. POR proteins in bacterial membranes were reconstituted with purified CYP2D6. Support of CYP2D6 was measured by metabolism of EOMCC (2H-1-benzopyran-3-carbonitrile,7-(ethoxy-methoxy)-2-oxo-(9Cl)), dextromethorphan and bufuralol. Michaelis constant (K(m)) and maximum velocity (V(max)) were determined in three triplicate experiments for each reaction; catalytic efficiency is expressed as V(max)/K(m). Compared with WT POR, disease-causing POR mutants A287P and R457H supported no detectable CYP2D6 activity with EOMCC, but A287P supported approximately 25% activity with dextromethorphan and bufuralol. Q153R had increased function with CYP2D6 (128% with EOMCC, 198% with dextromethorphan, 153% with bufuralol). A503V supported decreased CYP2D6 activity: 85% with EOMCC, 62% with dextromethorphan and 53% with bufuralol. POR variants have different effects depending on the substrate metabolized. Disease-causing POR mutations R457H and A287P had poor activities, suggesting that diminished drug metabolism should be considered in affected patients. The common A503V polymorphism impaired CYP2D6 activities with two commonly used drugs by 40-50%, potentially explaining some genetic variation in drug metabolism.


Central Compensation in Auditory Brainstem after Damaging Noise Exposure.
Noise exposure is one of the most common causes of hearing loss and peripheral damage to the auditory system. A growing literature suggests that the auditory system can compensate for peripheral loss through increased central neural activity. The current study sought to investigate the link between noise exposure, increases in central gain, synaptic reorganization, and auditory function. All axons of the auditory nerve project to the cochlear nucleus, making it a requisite nucleus for sound detection. As the first synapse in the central auditory system, the cochlear nucleus is well positioned to respond plastically to loss of peripheral input. To investigate noise-induced compensation in the central auditory system, we measured auditory brainstem responses (ABRs) and auditory perception and collected tissue from mice exposed to broadband noise. Noise-exposed mice showed elevated ABR thresholds, reduced ABR wave 1 amplitudes, and spiral ganglion neuron loss. Despite peripheral damage, noise-exposed mice were hyperreactive to loud sounds and showed nearly normal behavioral sound detection thresholds. Ratios of late ABR peaks (2-4) relative to the first ABR peak indicated that brainstem pathways were hyperactive in noise-exposed mice, while anatomical analysis indicated there was an imbalance between expression of excitatory and inhibitory proteins in the ventral cochlear nucleus. The results of the current study suggest that a reorganization of excitation and inhibition in the ventral cochlear nucleus may drive hyperactivity in the central auditory system. This increase in central gain can compensate for peripheral loss to restore some aspects of auditory function.


Association between objectively measured sleep duration and physical function in community-dwelling older adults.
The aim of this research was to investigate the relationships between objectively measured sleep duration and physical function in older adults. We recruited community-dwelling older adults aged ≥ 60 years old in Taipei City, Taiwan. Sleep duration was measured with accelerometers and recorded as the total hours of sleep per night for each participant. The following physical functions were assessed: 1) grip strength (measured by handgrip dynamometer), 2) balance (1-leg standing test), 3) lower body strength (5-timed chair stand), 4) basic mobility (timed up and go test), 5) gait speed (5-m walk test). The relationships between sleep duration and physical function outcomes were analyzed using generalized additive models, controlling for objectively measured sedentary behavior and moderate-to-vigorous physical activity, and other sociodemographic variables. A total of 121 older adults (men = 28.9%; mean age = 70.0 ± 5.0 years) was included in this study. A positive association of sleep duration with grip strength was found after adjusting for covariates (P = .005). No significant associations were observed between sleep duration and the other physical function outcomes. For older adults, lengthening their sleep duration may be helpful to enhance the grip strength. This result has implications for improving their health by targeting better performance in specific physical functions. Further studies of sleep duration and physical function among older adults should investigate the underlying mechanisms.


The application of fluorescence techniques in meningioma surgery-a review.
Surgical resections of meningiomas, the most common intracranial tumor in adults, can only be curative if radical resection is achieved. Potentially, the extent of resection could be improved, especially in complex and/or high-grade meningiomas by fluorescence-guided surgery using 5-aminolevulinic acid (5-ALA), indocyanine green (ICG), or fluorescein. This review aims to summarize and evaluate these fluorescence-guided meningioma surgery techniques. PubMed and Embase were searched for relevant articles. Additionally, we checked reference lists for further studies. Forty-eight articles were included in the final analysis. 5-ALA fluoresced with varying sensitivity and selectivity in meningiomas and in invaded bone and dura mater. Although ICG was mainly applied for video angiography, one report shows tumor fluorescence 18-28 h post-ICG injection. Lastly, the use of fluorescein could aid in the identification of tumor remnants; however, detection of dural tail is highly questionable. Fluorescence-guided meningioma surgery should be a reliable, highly specific, and sensitive technique. Despite numerous studies reporting the use of fluorescent dyes, currently, there is no evidence that these tools improve the radical resection rate and long-term recurrence-free outcome in meningioma surgery without neurological deficits. Evidence regarding the effectiveness and increased safety of resection after the application of these fluorophores is currently lacking. Future research should focus on the development of a meningioma-targeted, highly sensitive, and specific fluorophore.


8-OxoG retards the activity of the ligase III/XRCC1 complex during the repair of a single-strand break, when present within a clustered DNA damage site.
Ionising radiation produces clustered DNA damage. Recent studies have established that the efficiency of excision of a lesion within clustered damage sites is reduced. This study presents evidence that the repair of clustered DNA damage is compromised, relative to that of the isolated lesions, since the lifetime of both lesions is extended by up to eight fold. Simple clustered damage sites, comprised of a single-strand break, one or five bases 3' or 5' to 8-oxoG on the opposite strand, were synthesised in oligonucleotides and repair carried out in XRS5 nuclear extracts. The rate of repair of the single-strand break within these clustered damage sites is reduced, mainly due to inhibition of the DNA ligase III/XRCC1 complex. The single-strand break, present as an isolated lesion, is repaired by short-patch base excision repair, however the mechanism of repair of the single-strand break within the clustered damage site is asymmetric. When the lesions are 5' to each other, the single-strand break is rejoined by short-patch repair whereas the rejoining of the single-strand break occurs by long-patch type repair when the lesions are 3' to one another. The retardation of DNA ligase III/XRCC1 complex, following addition of one base, is responsible for the initiation of long-patch base excision repair when the lesions are 3' to each other. The lesions within the cluster are processed sequentially, the single-strand break being repaired before excision of 8-oxoG, limiting the formation of double-strand breaks to <2%. Stalled processing of clustered DNA damage is suggested to have implications for mutation induction by radiation.


Infusing Technology Into Perinatal Home Visitation in the United States for Women Experiencing Intimate Partner Violence: Exploring the Interpretive Flexibility of an mHealth Intervention.
Intimate partner violence (IPV) is common during pregnancy and the postpartum. Perinatal home visitation provides favorable conditions in which to identify and support women affected by IPV. However, the use of mHealth for delivering IPV interventions in perinatal home visiting has not been explored. Our objective was to conduct a nested qualitative interpretive study to explore perinatal home visitors' and women's perceptions and experiences of the Domestic Violence Enhanced Home Visitation Program (DOVE) using mHealth technology (ie, a computer tablet) or a home visitor-administered, paper-based method. We used purposive sampling, using maximum variation, to select women enrolled in a US-based randomized controlled trial of the DOVE intervention for semistructured interviews. Selection criteria were discussed with the trial research team and 32 women were invited to participate. We invited 45 home visitors at the 8 study sites to participate in an interview, along with the 2 DOVE program designers. Nonparticipant observations of home visits with trial participants who chose not to participate in semistructured interviews were undertaken. We conducted 51 interviews with 26 women, 23 home visiting staff at rural and urban sites, and the 2 DOVE program designers. We conducted 4 nonparticipant observations. Among 18 IPV-positive women, 7 used the computer tablet and 11 used the home visitor method. Among 8 IPV-negative women, 7 used the home visitor method. The computer tablet was viewed as a safe and confidential way for abused women to disclose their experiences without fear of being judged. The meanings that the DOVE technology held for home visitors and women led to its construction as either an impersonal artifact that was an impediment to discussion of IPV or a conduit through which interpersonal connection could be deepened, thereby facilitating discussion about IPV. Women's and home visitors' comfort with either method of screening was positively influenced by factors such as having established trust and rapport, as well as good interpersonal communication. The technology helped reduce the anticipated stigma associated with disclosing abuse. The didactic intervention video was a limiting feature, as the content could not be tailored to accommodate the fluidity of women's circumstances. Users and developers of technology-based IPV interventions need to consider the context in which they are being embedded and the importance of the patient-provider relationship in promoting behavior change in order to realize the full benefits. An mHealth approach can and should be used as a tool for initiating discussion about IPV, assisting women in enhancing their safety and exploring help-seeking options. However, training for home visitors is required to ensure that a computer tablet is used to complement and enhance the therapeutic relationship. Clinicaltrials.gov NCT01688427; https://clinicaltrials.gov/ct2/show/NCT01688427 (Archived by WebCite at http://www.webcitation.org/6limSWdZP).


The distribution and morphological characteristics of catecholaminergic cells in the diencephalon and midbrain of the bottlenose dolphin (Tursiops truncatus).
The present study describes the distribution and cellular morphology of catecholaminergic neurons in the diencephalon and midbrain of the bottlenose dolphin (Tursiops truncatus). Tyrosine hydroxylase immunohistochemistry was used to visualize these putatively dopaminergic neurons. The standard A1-A17, C1-C3, nomenclature is used for expediency; however, the neuroanatomical names of the various nuclei have also been given. Dolphins exhibit certain tyrosine hydroxylase immunoreactive (TH-ir) catecholaminergic neuronal groups in the midbrain (A8, A9, A10) and diencephalon (A11, A12, A14), however, no neuronal clusters clearly corresponding to the A13 and A15 groups could be identified. The subdivisions of these neuronal groups are in general agreement with those of other mammals, but there is a high degree of species specificity. First, three TH-ir neuronal groups not identified in other species were found: in the ventral lateral peri-aqueductal gray matter, posterior dorsal hypothalamus, and rostral mesencephalic raphe. Second, the normal components of the substantia nigra (A9 or pars compacta, A9 lateral or pars lateralis, A9 ventral or pars reticulata) were extremely cell sparse, but there was a substantial expansion of the A9 medial and A10 lateral subdivisions forming an impressive 'ventral wing' in the posterior substantia nigra. The findings of this and previous studies suggest a distinct evolutionary trend occurring in the neuromodulatory systems in mammals. The results are discussed in relation to motor control, thermoregulation, unihemispheric sleep, and dolphin cognition.


Characterization of determinants of ligand binding to the nicotinic acid receptor GPR109A (HM74A/PUMA-G).
The G-protein-coupled receptor GPR109A (HM74A/PUMA-G) has recently been shown to function as a receptor for nicotinic acid (niacin) and to mediate its antilipolytic effects. Nicotinic acid is able to strongly raise plasma levels of high-density lipoprotein cholesterol, a property that distinguishes nicotinic acid from other lipid-lowering drugs. To investigate the structural determinants of GPR109A ligand binding, we performed site-directed mutagenesis of putative ligand binding residues combined with generation of chimeric receptors consisting of GPR109A and its close relative GPR109B, which does not bind nicotinic acid. We could identify Asn86/Trp91 [transmembrane helix (TMH) 2/extracellular loop (ECL) 1], Arg111 (TMH3), Ser178 (ECL2), Phe276 (TMH7), and Tyr284 (TMH7) as amino acid residues critical for binding of nicotinic acid. Together with data from molecular modeling studies, our data suggest that the ligand binding pocket for nicotinic acid of GPR109A is distinct from that of most other group A receptors. Although Arg111 at TMH3 serves as the basic anchor point for the carboxylate ligands, the ring system of nicotinic acid is embedded between Trp91 at the junction TMH2/ECL1 and Phe276/Tyr284 at TMH7. The heterocyclic ring is also bound to Ser178 at ECL2 via an H-bond. These data will facilitate the design of new antidyslipidemic drugs acting via GPR109A.


Defensive movements evoked by air puff in monkeys.
Electrical stimulation of two connected cortical areas in the monkey brain, the ventral intraparietal area (VIP) in the intraparietal sulcus and the polysensory zone (PZ) in the precentral gyrus, evokes a specific set of movements. In one interpretation, these movements correspond to those typically used to defend the body from objects that are near, approaching, or touching the skin. The present study examined the movements evoked by a puff of air aimed at various locations on the face and body of fascicularis monkeys to compare them to the movements evoked by stimulation of VIP and PZ. The air-puff-evoked movements included a movement of the eyes from any initial position toward a central region and a variety of stereotyped facial, shoulder, head, and arm movements. These movements were similar to those reported on stimulation of VIP and PZ. One difference between the air-puff-evoked movements and those evoked by stimulation of VIP and PZ is that the air puff evoked an initial startle response (a bilaterally symmetric spike in muscle activity) followed by a more sustained, lateralized response, specific to the site of the air puff. In contrast, stimulation of VIP and PZ evoked mainly a sustained, lateralized response, specific to the site of the receptive fields of the stimulated neurons. We speculate that VIP and PZ may contribute to the control of defensive movements, but that they may emphasize the more spatially specific reactions that occur after startle.


Improving precision by adjusting for prognostic baseline variables in randomized trials with binary outcomes, without regression model assumptions.
In randomized clinical trials with baseline variables that are prognostic for the primary outcome, there is potential to improve precision and reduce sample size by appropriately adjusting for these variables. A major challenge is that there are multiple statistical methods to adjust for baseline variables, but little guidance on which is best to use in a given context. The choice of method can have important consequences. For example, one commonly used method leads to uninterpretable estimates if there is any treatment effect heterogeneity, which would jeopardize the validity of trial conclusions. We give practical guidance on how to avoid this problem, while retaining the advantages of covariate adjustment. This can be achieved by using simple (but less well-known) standardization methods from the recent statistics literature. We discuss these methods and give software in R and Stata implementing them. A data example from a recent stroke trial is used to illustrate these methods.


[Blind source separation for fMRI signals using a new independent component analysis algorithm and principal component analysis].
The application of independent component analysis (ICA) to the functional magnetic resonance imaging (fMRI) data can separate many independent sources. But in the processing there are two difficulties: (1) the data of the fMRI is usually on a large scale, so the computing is time-consuming; (2) we cannot avoid the errors for too heavy computational load, this brings many troubles. Thus we think of reducing the data. In this article we used the standard information theoretic methods to estimate the number of the sources and used the principal component analysis (PCA) to reduce the data. By this process, we estimated the number of the sources and reduced the data successfully; Then we applied the ICA algorithm to the reduced fMRI data; this method raised the speed of operation. After application of the new ICA algorithm and another algorithm (FastICA) to the fMRI data, a comparison was made. The results show that the new algorithm can separate the fMRI data fast and effectively and it is superior to the FastICA on the accuracy of estimating the temporal dynamics of activations.


Reproducibility of temporomandibular joint clicking.
To investigate the stability of temporomandibular joint (TMJ) clicking over a 10-day period and the effect of different open/close velocities on sound amplitude and power spectra in a group of subjects with subjectively stable unilateral clicking during the 3 months preceding the recordings. Ten volunteers were recorded with a self-developed microcomputer-based system used in a previous study on asymptomatic subjects. The recordings were performed during 4 different sessions at 3 different open/close rates in each session. The subjective sound intensity was measured with a 100-mm visual analog scale (VAS). The VAS scores, the maximum amplitudes, and the power spectra of the signals were tested for statistical differences among the different open/close rates and over the sessions. The reliability of measurements was also calculated. The maximum amplitude and the power spectra of the TMJ clicking varied between subjects in a broad range that differed from those reported for asymptomatic subjects. No statistically significant differences were found within subjects for the subjective VAS scores for the maximum signal amplitudes or for the power spectrum parameters among the open/close rates and over the 4 sessions. For all 3 open/close rates and for the 4 sessions, a good to excellent reliability of measurements was determined, the values of r being mostly over 0.75. Within the limits of the experiment, TMJ clicking was subjectively and objectively stable over a period of 10 days. Therefore, the constant subjective perception of sound intensity was supported by the objective measurements.


Hyperplane-Approximation-Based Method for Many-Objective Optimization Problems with Redundant Objectives.
For a many-objective optimization problem with redundant objectives, we propose two novel objective reduction algorithms for linearly and, nonlinearly degenerate Pareto fronts. They are called LHA and NLHA respectively. The main idea of the proposed algorithms is to use a hyperplane with non-negative sparse coefficients to roughly approximate the structure of the PF. This approach is quite different from the previous objective reduction algorithms that are based on correlation or dominance structure. Especially in NLHA, in order to reduce the approximation error, we transform a nonlinearly degenerate Pareto front into a nearly linearly degenerate Pareto front via a power transformation. In addition, an objective reduction framework integrating a magnitude adjustment mechanism and a performance metric σ* are also proposed here. Finally, to demonstrate the performance of the proposed algorithms, comparative experiments are done with two correlation-based algorithms, LPCA and NLMVUPCA, and with two dominance-structure-based algorithms, PCSEA and greedy δ- MOSS, on three benchmark problems: DTLZ5(I,M), MAOP(I,M), and WFG3(I,M). Experimental results show that the proposed algorithms are more effective.


Ozone-induced respiratory illness during the repair of a portland cement kiln.
Workers at a portland cement plant had experienced acute respiratory and eye irritation when performing maintenance inside a kiln. These episodes were associated with a bleach-like odor, which was only reported during maintenance operations. An industrial hygiene investigation was conducted to determine the cause of the illness. While workers replaced refractory brick inside the kiln, air samples were collected for chlorine, sulfur dioxide, inorganic acid, ozone, and dust. After the rebricking was completed and all the workers had exited the kiln, its electrostatic precipitator was reduced to half power and the induced-draft (ID) fan was turned off to recreate conditions present during illness episodes. Chlorine, inorganic acid, and ozone were not detected, and only trace concentrations of sulfur dioxide were detected while workers were inside the kiln. However, when conditions present during previous episodes were recreated, the bleach-like odor was soon evident. Chlorine was not detected, but 0.09 to 0.11 ppm of ozone was measured at the discharge end of the kiln, and 4.5 ppm was measured at the inlet end. Within a half hour after the electrostatic precipitator was turned off and the ID fan was turned on, the ozone concentrations decreased to background levels of 0.02-0.03 ppm. Somewhat lower ozone exposures may have occurred during previous kiln maintenance operations due to more open access portals, but previous episodes of eye and respiratory irritation were probably caused when ozone, generated by the electrostatic precipitator, back-drafted into the kiln after the ID fan was turned off.


Chemical and sensory characterisation of Sangiovese red wines: comparison between biodynamic and organic management.
The effects of biodynamic production practices on composition and sensory attributes of Sangiovese wines were examined for 2 years (2009 and 2010) in a vineyard that was converted from organic (ORG) to biodynamic (BDN) viticulture. During the first year (2009), the BDN wines were characterised by low alcohol strength, colour intensity, total polyphenols, monomeric anthocyanins and catechin. Conversely, the second year BDN wines differed from the organic wines in terms of total polyphenols and phenolic compounds, including polymeric pigments, co-pigmentation, tannins and iron-reactive polyphenols. The effect of management practices, harvest and their interaction was analysed for each compound. Positive interaction was observed for total acidity, volatile acidity, cyanidin-3-glucoside, protocatechuic acid, (+)-catechin, quercetin and trans-resveratrol. ORG wine initially showed a more complex aroma profile; however, the differences were almost indistinguishable during the second year. Trained panellists highlighted differences in colour intensity between ORG and BDN wines although no preference was found by consumers. The concentrations of ochratoxin A and biogenic amines were far below the health-hazardous threshold.


Pharmacokinetics of tramadol enantiomers and their respective phase I metabolites in relation to CYP2D6 phenotype.
Our objective was to evaluate the effect of CYP2D6 phenotype in the enantioselective metabolism of tramadol in Spanish healthy human volunteers. A single oral 100mg dose of racemic tramadol was administered to five subjects who were poor metabolizers (PMs) and 19 subjects who were extensive metabolizers (EMs), whose phenotypes were determined by the use of the racemic tramadol metabolic rate. The pharmacokinetic parameters were estimated from plasma concentrations of the enantiomers of tramadol and their main phase I metabolites, O-desmethyltramadol (M1) and N-desmethyltramadol (M2). Epinephrine plasma concentrations were also determinated. The plasma concentrations of both tramadol enantiomers were consistently higher in PMs than in EMs of CYP2D6, with 1.98- and 1.74-fold differences in the mean area under the plasma concentration-time curves (AUC), respectively. The values for oral clearance of (+)- and (--)-tramadol were 1.91- and 1.71-fold greater in PMs, which were related to differences in both O-desmethylation and N-desmethylation in the two CYP2D6 metabolizer phenotypes. The mean AUC values of (+)-M1 and (--)-M1 were 4.33- and 0.89-fold greater in EMs, and it was related to similar differences in the formation rate constant. On the other hand, the differences were 7.40- and 8.69-fold greater in PMs for M2 enantiomers due to the involvement of CYP2D6 in their subsequent biotransformation. The time course of epinephrine systemic concentrations was completely different between both groups of metabolizers. In EMs plasma concentrations of epinephrine increased after tramadol administration whereas in PMs no effect was observed. The polymorphic CYP2D6 appears to be a major enzyme involved in the metabolism of tramadol enantiomers. The N-desmethylation pathway was indirectly affected by CYP2D6 phenotypic differences. Epinephrine showed a good correlation with the pharmacokinetics of the opioid component of tramadol, (+)-M1 and was found to be useful for its pharmacodynamic profiling.


Lead, cadmium and cobalt (Pb, Cd, and Co) leaching of glass-clay containers by pH effect of food.
Recent studies have shown that handcrafted glass-clay containers are a health risk because they can be contaminated by heavy metals, which can be transferred to food, thus reaching the human body to potentially cause illness. Therefore, in the present work, we evaluate the leaching of lead, cadmium, and cobalt from glass-clay containers into two types of food: tomato sauce (salsa), and chickpea puree. The containers were obtained from four regions in the Mexican state of Hidalgo. Repetitive extractions from the containers were carried out to quantify the leaching of the heavy metals into the salsa, the chickpea puree, and acetic acid using the technique proposed by the USFDA. The results show that greater use of the containers leads to more leaching of heavy metals into both types of food and into the acetic acid, with the greatest metal extraction recorded for the Ixmiquilpan vessels. These results indicate that the metals present in the glass-clay containers leach into the food and that increased reuse increases the risk to the people who use them in food preparation.


Effect of attentional load on audiovisual speech perception: evidence from ERPs.
Seeing articulatory movements influences perception of auditory speech. This is often reflected in a shortened latency of auditory event-related potentials (ERPs) generated in the auditory cortex. The present study addressed whether this early neural correlate of audiovisual interaction is modulated by attention. We recorded ERPs in 15 subjects while they were presented with auditory, visual, and audiovisual spoken syllables. Audiovisual stimuli consisted of incongruent auditory and visual components known to elicit a McGurk effect, i.e., a visually driven alteration in the auditory speech percept. In a Dual task condition, participants were asked to identify spoken syllables whilst monitoring a rapid visual stream of pictures for targets, i.e., they had to divide their attention. In a Single task condition, participants identified the syllables without any other tasks, i.e., they were asked to ignore the pictures and focus their attention fully on the spoken syllables. The McGurk effect was weaker in the Dual task than in the Single task condition, indicating an effect of attentional load on audiovisual speech perception. Early auditory ERP components, N1 and P2, peaked earlier to audiovisual stimuli than to auditory stimuli when attention was fully focused on syllables, indicating neurophysiological audiovisual interaction. This latency decrement was reduced when attention was loaded, suggesting that attention influences early neural processing of audiovisual speech. We conclude that reduced attention weakens the interaction between vision and audition in speech.


The importance of morphemic awareness to reading achievement and the potential of signing morphemes to supporting reading development.
The ability to access and understand the meaning of multi-morphemic words is essential for age-appropriate literacy growth as well as for achievement in other participants, such as science and social studies, which are so print-dependent. This paper provides a theoretical basis for focusing on the morphology of English when teaching students who are deaf or hard of hearing to read through a review of the literature on the role of morphology in reading for both hearing students and those with a hearing loss. In addition, the authors review the empirical literature on Signing Exact English (SEE), a system of signing English constructed in which the morphology of words is made visible to children who might not be able to hear them. The authors propose that students' use of SEE can provide a bridge to developing the morphemic awareness so necessary for age-appropriate reading development and achievement.


Cardiomyocyte hepcidin: From intracellular iron homeostasis to physiological function.
Cellular iron is required for the utilization of oxygen in the cell. Iron in iron-sulfur and heme groups is required for electron transfer and oxygen activation in oxidative phosphorylation, while labile free iron is required for oxygen activation by dioxygenases, and as a catalyst for redox signaling. At the same time, this reactivity with oxygen underpins the production of cell-damaging free radicals in the presence of excess iron. Because the cardiac cell is a major site of oxygen flux, it requires tight control of intracellular iron levels. Until recently, such control was thought to be mediated predominantly by the action of iron regulatory proteins. However, new evidence reveals that cardiomyocyte hepcidin is indispensable for the control of intracellular iron levels, normal metabolism and heart function. This new evidence highlights the need for better understanding of the regulation of cardiomyocyte hepcidin in health and disease.


Inhibition of ADP ribosylation prevents and cures helicobacter-induced gastric preneoplasia.
Gastric adenocarcinoma develops as a consequence of chronic inflammation of the stomach lining that is caused by persistent infection with the bacterium Helicobacter pylori. Gastric carcinogenesis progresses through a sequence of preneoplastic lesions that manifest histologically as atrophic gastritis, intestinal metaplasia, and dysplasia. We show here in several preclinical models of Helicobacter-induced atrophic gastritis, epithelial hyperplasia, and metaplasia that the inhibition of ADP ribosylation by the small-molecule inhibitor PJ34 not only prevents the formation of gastric cancer precursor lesions, but also efficiently reverses preexisting lesions. PJ34 exerts its chemopreventive and therapeutic effects by impairing Helicobacter-specific T-cell priming and T(H)1 polarization in the gut-draining mesenteric lymph nodes. The subsequent infiltration of pathogenic T cells into the gastric mucosa and the ensuing gastric T cell-driven immunopathology are prevented efficiently by PJ34. Our data indicate that PJ34 directly suppresses T-cell effector functions by blocking the IFN-gamma production of mesenteric lymph node T cells ex vivo. Upon exposure to PJ34, purified T cells failed to synthesize ADP-ribose polymers and to activate the transcription of genes encoding IFN-gamma, interleukin 2, and the interleukin 2 receptor alpha chain in response to stimuli such as CD3/CD28 cross-linking or phorbol 12-myristate 13-acetate/ionomycin. The immunosuppressive and chemoprotective effects of PJ34 therefore result from impaired T-cell activation and T(H)1 polarization, and lead to the protection from preneoplastic gastric immunopathology. In conclusion, ADP-ribosylating enzymes constitute novel targets for the treatment of Helicobacter-associated gastric lesions predisposing infected individuals to gastric cancer and may also hold promise for the treatment of other T cell-driven chronic inflammatory conditions and autoimmune pathologies.


Crossing the Red Sea: phylogeography of the hamadryas baboon, Papio hamadryas hamadryas.
The hamadryas baboon (Papio hamadryas hamadryas) is found both in East Africa and western Arabia and is the only free-ranging nonhuman primate in Arabia. It has been hypothesized that hamadryas baboons colonized Arabia in the recent past and were possibly even transported there by humans. We investigated the phylogeography of hamadryas baboons by sequencing a portion of the control region of mtDNA in 107 baboons from four Saudi Arabian populations and combing these data with published data from Eritrean (African) P. h. hamadryas. Analysis grouped sequences into three distinct clades, with clade 1 found only in Arabia, clade 3 found only in Africa, but clade 2 found in both Arabian and African P. h. hamadryas and also in the olive baboon, P. h. anubis. Patterns of variation within Arabia are neither compatible with the recent colonization of Arabia, implying that baboons were not transported there by humans, nor with a northerly route of colonization of Arabia. We propose that hamadryas baboons reached Arabia via land bridges that have formed periodically during glacial maxima at the straits of Bab el Mandab in the southern Red Sea. We suggest that the genetic differentiation of Arabian from African populations suggests that Arabian populations have a higher conservation status than recognized previously.


Plasmon transmission through excitonic subwavelength gaps.
We study the transfer of electromagnetic energy across a subwavelength gap separating two co-axial metal nanorods. In the absence of spacer in the gap separating the rods, the system exhibits strong coupling behavior between longitudinal plasmons in the two rods. The nature and magnitude of this coupling are studied by varying various geometrical parameters. As a function of frequency, the transmission is dominated by a split longitudinal plasmon peak. The two hybrid modes are the dipole-like "bonding" mode characterized by a peak intensity in the gap and a quadrupole-like "antibonding" mode whose amplitude vanishes at the gap center. When the length of one rod is varied, this mode spectrum exhibits the familiar anti-crossing behavior that depends on the coupling strength determined by the gap width. When off-resonant 2-level emitters are placed in the gap, almost no effect on the frequency dependent transmission is observed. In contrast, when the molecular system is resonant with the plasmonic line shape, the transmission is strongly modified, showing characteristics of strong exciton-plasmon coupling. Most strongly modified is the transmission near the lower frequency "bonding" plasmon mode. The presence of resonant molecules in the gap affects not only the molecule-field interaction but also the spatial distribution of the field intensity and the electromagnetic energy flux across the junction.


The Cutaneous Microbiome and Wounds: New Molecular Targets to Promote Wound Healing.
The ecological community of microorganisms in/on humans, termed the microbiome, is vital for sustaining homeostasis. While culture-independent techniques have revealed the role of the gut microbiome in human health and disease, the role of the cutaneous microbiome in wound healing is less defined. Skin commensals are essential in the maintenance of the epithelial barrier function, regulation of the host immune system, and protection from invading pathogenic microorganisms. In this review, we summarize the literature derived from pre-clinical and clinical studies on how changes in the microbiome of various acute and chronic skin wounds impact wound healing tissue regeneration. Furthermore, we review the mechanistic insights garnered from model wound healing systems. Finally, in the face of growing concern about antibiotic-resistance, we will discuss alternative strategies for the treatment of infected wounds to improve wound healing and outcomes. Taken together, it has become apparent that commensals, symbionts, and pathogens on human skin have an intimate role in the inflammatory response that highlights several potential strategies to treat infected, non-healing wounds. Despite these promising results, there are some contradictory and controversial findings from existing studies and more research is needed to define the role of the human skin microbiome in acute and chronic wound healing.


MLH1 protects from resistance acquisition by the histone deacetylase inhibitor trichostatin A in colon tumor cells.
The antineoplastic activity of HDAC inhibitors is an unquestionable property of these compounds, but recent studies in tumor cells have revealed the potential of HDAC inhibitors (e.g., suberoylanilide hydroxamic acid SAHA, valproic acid VPA) to cause acquisition of HDAC inhibitor resistance. We report that trichostatin A (TSA), an HDAC inhibitor structurally related to SAHA, causes the acquisition of multidrug resistance transporter-independent and irreversible 3-fold resistance to TSA in MLH1-deficient (absent MLH1 protein expression) but not in MLH1-proficient (expressing MLH1 protein) HCT116 colon tumor cells. This MLH1-deficient subline selected for TSA resistance by stepwise exposures to increasing TSA concentrations exhibited failure in the accumulation of acetylated histones, in p21 induction, and in apoptosis activation. These are cellular responses normally seen in tumor cells treated with HDAC inhibitors. Whereas the absence of acetyl-histone accumulation did not correlate with altered HDAC activity, the absence of apoptosis correlated with reduced expression of (pro-apoptotic) Bax. This TSA-resistant subline was cross-resistant to SAHA and VPA but not to 'classic' non-HDAC inhibitor-type anticancer agents such as docetaxel and doxorubicin. These herein presented results expand on a previous study reporting HDAC inhibitor resistance acquisition by SAHA which was independent of the MLH1 expression status. Taken together, the present study identifies TSA, besides SAHA and VPA, as another potential causative of HDAC inhibitor resistance acquisition specifically in MLH1-deficient HCT116 colon tumor cells, and it reveals a possible function of MLH1 protein in protecting colon tumor cells from resistance acquisition by TSA.


Emergency management in patients with haemophilia A and inhibitors on prophylaxis with emicizumab: AICE practical guidance in collaboration with SIBioC, SIMEU, SIMEUP, SIPMeL and SISET.
Emicizumab has been approved in several countries for regular prophylaxis in patients with congenital haemophilia A and FVIII inhibitors because it substantially reduces their bleeding risk and improves quality of life. However, although significantly less frequent, some breakthrough bleeds may still occur while on emicizumab, requiring treatment with bypassing or other haemostatic agents. Thrombotic complications have been reported with the associated use of activated prothrombin complex concentrates. In addition, when surgery/invasive procedures are needed while on emicizumab, their management requires multidisciplinary competences and direct supervision by experts in the use of this agent. Given this, and in order to expand the current knowledge on the use of emicizumab and concomitant haemostatic agents, and reduce the risk of complications in this setting, the Italian Association of Haemophilia Centres (AICE) here provides guidance on the management of breakthrough bleeds and surgery in emergency situations in patients with haemophilia A and inhibitors on emicizumab prophylaxis. This paper has been shared with other National Scientific Societies involved in the field.


An HER2-Displaying Virus-Like Particle Vaccine Protects from Challenge with Mammary Carcinoma Cells in a Mouse Model.
Human epidermal growth factor receptor-2 (HER2) is upregulated in 20% to 30% of breast cancers and is a marker of a poor outcome. Due to the development of resistance to passive immunotherapy with Trastuzumab, active anti-HER2 vaccination strategies that could potentially trigger durable tumor-specific immune responses have become an attractive research area. Recently, we have shown that budded virus-like particles (VLPs) produced in Sf9 insect cells are an ideal platform for the expression of complex membrane proteins. To assess the efficacy of antigen-displaying VLPs as active cancer vaccines, BALB/c mice were immunized with insect cell glycosylated and mammalian-like glycosylated HER2-displaying VLPs in combination with two different adjuvants and were challenged with HER2-positive tumors. Higher HER2-specific antibody titers and effector functions were induced in mice vaccinated with insect cell glycosylated HER2 VLPs compared to mammalian-like glycosylated counterparts. Moreover, insect cell glycosylated HER2 VLPs elicited a protective effect in mice grafted with HER2-positive mammary carcinoma cells. Interestingly, no protection was observed in mice that were adjuvanted with Poly (I:C). Here, we show that antigen-displaying VLPs produced in Sf9 insect cells were able to induce robust and durable immune responses in vivo and have the potential to be utilized as active cancer vaccines.


Randomized controlled trial comparing the combination of a polymeric membrane dressing plus negative pressure wound therapy against negative pressure wound therapy alone: The WICVAC study.
Negative pressure wound therapy (NPWT) is the treatment of choice for chronic wounds; yet, it is associated with considerable workload. Prompted by its nonadhesive and wound-healing properties, this study investigated the effect of an additional polymeric membrane interface dressing (PMD; PolyMem WIC) in NPWT. From October 2011 to April 2013, 60 consecutive patients with chronic leg wounds or surgical site infections after revascularization of lower extremities were randomly allocated to either treatment with conventional NPWT (control arm) or NPWT with an additional PMD (intervention arm). The primary outcome was wound healing achieved within 30 days, the secondary endpoints included: number of days between dressing changes, wound-related pain, cost efficiency, and occurrence of adverse events (ClinTrials.gov Identifier: NCT02399722). Forty-seven patients completed follow-up. No difference in wound healing was observed (p > 0.05) between both study arms. The additional PMD allowed significantly longer wearing times (days) between dressing changes (intervention: 8.8 ± 0.5, control: 4.8 ± 0.2; p < 0.001). Pain was slightly higher in patients randomized to NPWT alone (VAS score: 4.8 ± 2.9) compared to NPWT + PMD (VAS score: 3.0 ± 2.9, p = 0.063). No wound infections were observed. Costs were reduced by 34% per patient in the intervention arm. These results suggest that the combination of NPWT and an additional interface PMD is a safe and economic method for the treatment of chronic wounds, which requires significantly fewer dressing changes for a comparable wound healing.


Relationships Among Meeting Physical-Activity Guidelines and Health Risk Behaviors.
Young adults have the highest participation in physical activity but also have the highest incidence rates of binge drinking, cigarette smoking, and smokeless tobacco use. We examined these factors to determine whether there are relationships among physical activity and health risk behaviors. We conducted correlation and χ² analyses using the American College Health Association-National College Health Assessment fall 2009 data set (N = 34,208) to examine the relationship among meeting physical-activity guidelines, binge drinking, and tobacco use among survey participants. The data suggest a positive relationship between meeting physical-activity guidelines and binge drinking, with the strongest relationship between those reporting binge drinking 4 times in a 2-week period. Meeting physical-activity guidelines was negatively associated with cigarette use but positively associated with all other types of tobacco use. Associations between physical activity and binge-drinking episodes indicate a need to address the relationship between heavy drinking and alcohol dependence and physical-activity behavior patterns. Further studies should examine relationships between physical activity and binge drinking in other age groups. Results also suggest the need to examine differing associations between physical activity and types of tobacco use.


Double-slit experiment with single wave-driven particles and its relation to quantum mechanics.
In a thought-provoking paper, Couder and Fort [Phys. Rev. Lett. 97, 154101 (2006)] describe a version of the famous double-slit experiment performed with droplets bouncing on a vertically vibrated fluid surface. In the experiment, an interference pattern in the single-particle statistics is found even though it is possible to determine unambiguously which slit the walking droplet passes. Here we argue, however, that the single-particle statistics in such an experiment will be fundamentally different from the single-particle statistics of quantum mechanics. Quantum mechanical interference takes place between different classical paths with precise amplitude and phase relations. In the double-slit experiment with walking droplets, these relations are lost since one of the paths is singled out by the droplet. To support our conclusions, we have carried out our own double-slit experiment, and our results, in particular the long and variable slit passage times of the droplets, cast strong doubt on the feasibility of the interference claimed by Couder and Fort. To understand theoretically the limitations of wave-driven particle systems as analogs to quantum mechanics, we introduce a Schrödinger equation with a source term originating from a localized particle that generates a wave while being simultaneously guided by it. We show that the ensuing particle-wave dynamics can capture some characteristics of quantum mechanics such as orbital quantization. However, the particle-wave dynamics can not reproduce quantum mechanics in general, and we show that the single-particle statistics for our model in a double-slit experiment with an additional splitter plate differs qualitatively from that of quantum mechanics.


Effector biology of plant-associated organisms: concepts and perspectives.
Every plant is closely associated with a variety of living organisms. Therefore, deciphering how plants interact with mutualistic and parasitic organisms is essential for a comprehensive understanding of the biology of plants. The field of plant-biotic interactions has recently coalesced around an integrated model. Major classes of molecular players both from plants and their associated organisms have been revealed. These include cell surface and intracellular immune receptors of plants as well as apoplastic and host-cell-translocated (cytoplasmic) effectors of the invading organism. This article focuses on effectors, molecules secreted by plant-associated organisms that alter plant processes. Effectors have emerged as a central class of molecules in our integrated view of plant-microbe interactions. Their study has significantly contributed to advancing our knowledge of plant hormones, plant development, plant receptors, and epigenetics. Many pathogen effectors are extraordinary examples of biological innovation; they include some of the most remarkable proteins known to function inside plant cells. Here, we review some of the key concepts that have emerged from the study of the effectors of plant-associated organisms. In particular, we focus on how effectors function in plant tissues and discuss future perspectives in the field of effector biology.


A new method of detecting changes in corneal health in response to toxic insults.
The size and arrangement of stromal collagen fibrils (CFs) influence the optical properties of the cornea and hence its function. The spatial arrangement of the collagen is still questionable in relation to the diameter of collagen fibril. In the present study, we introduce a new parameter, edge-fibrillar distance (EFD) to measure how two collagen fibrils are spaced with respect to their closest edges and their spatial distribution through normalized standard deviation of EFD (NSDEFD) accessed through the application of two commercially available multipurpose solutions (MPS): ReNu and Hippia. The corneal buttons were soaked separately in ReNu and Hippia MPS for five hours, fixed overnight in 2.5% glutaraldehyde containing cuprolinic blue and processed for transmission electron microscopy. The electron micrographs were processed using ImageJ user-coded plugin. Statistical analysis was performed to compare the image processed equivalent diameter (ED), inter-fibrillar distance (IFD), and EFD of the CFs of treated versus normal corneas. The ReNu-soaked cornea resulted in partly degenerated epithelium with loose hemidesmosomes and Bowman's collagen. In contrast, the epithelium of the cornea soaked in Hippia was degenerated or lost but showed closely packed Bowman's collagen. Soaking the corneas in both MPS caused a statistically significant decrease in the anterior collagen fibril, ED and a significant change in IFD, and EFD than those of the untreated corneas (p<0.05, for all comparisons). The introduction of EFD measurement in the study directly provided a sense of gap between periphery of the collagen bundles, their spatial distribution; and in combination with ED, they showed how the corneal collagen bundles are spaced in relation to their diameters. The spatial distribution parameter NSDEFD indicated that ReNu treated cornea fibrils were uniformly distributed spatially, followed by normal and Hippia. The EFD measurement with relatively lower standard deviation and NSDEFD, a characteristic of uniform CFs distribution, can be an additional parameter used in evaluating collagen organization and accessing the effects of various treatments on corneal health and transparency.


Double Jeopardy-Analyzing the Combined Effect of Age and Gender Stereotype Threat on Older Workers.
In this study we aim to analyze the combined effect of age-based and gender stereotype threat on work identity processes (and in particular on authenticity and organizational identification) and on work performance (self-rating performance). The research utilizes an ample sample of over fifty-year-old workers from diverse organizations in Italy. Using a person-centered approach four clusters of workers were identified: low in both age-based and gender stereotype threat (N = 4,689), high in gender and low in age-based stereotype threat (N = 1,735), high in age-based and low in gender stereotype threat (N = 2,013) and high in both gender and age-based stereotype threat (N = 758). Gender was significantly associated with these clusters and women were more frequently present in those groups with high gender stereotype threat. ANOVA results show that workers in the last two clusters score significantly lower in authenticity, organizational identification and self-rate performance. All in all, if ageism is undoubtedly problematic for older workers' identity processes, ageism and gender-stereotypes represent a double risk for women over fifty in the workplace. The analysis of the results can be beneficial both for the theoretical advancement and for the practical insights offered in the organizational and management field, where new policies of HR management can be elaborated, in order to value and to improve the workers experience.


AAAS Meeting Draws a Crowd.
The annual meeting of the AAAS, held in San Francisco on 14 to 19 January, attracted the largest turnout for several years. More than 6000 are estimated to have attended. The meeting, which was held in conjunction with the winter meeting of the American Physical Society and the annual meeting of the American Association of Physics Teachers, was reminiscent in some respects of the big AAAS annual gatherings of the late 1960s and early 1970s-even down to the presence of demonstrators, this year from the animal rights movement. Some highlights of the more than 250 sessions:


Gata3 targets Runx1 in the embryonic haematopoietic stem cell niche.
Runx1 is an important haematopoietic transcription factor as stressed by its involvement in a number of haematological malignancies. Furthermore, it is a key regulator of the emergence of the first haematopoietic stem cells (HSCs) during development. The transcription factor Gata3 has also been linked to haematological disease and was shown to promote HSC production in the embryo by inducing the secretion of important niche factors. Both proteins are expressed in several different cell types within the aorta-gonads-mesonephros (AGM) region, in which the first HSCs are generated; however, a direct interaction between these two key transcription factors in the context of embryonic HSC production has not formally been demonstrated. In this current study, we have detected co-localisation of Runx1 and Gata3 in rare sub-aortic mesenchymal cells in the AGM. Furthermore, the expression of Runx1 is reduced in Gata3 -/- embryos, which also display a shift in HSC emergence. Using an AGM-derived cell line as a model for the stromal microenvironment in the AGM and performing ChIP-Seq and ChIP-on-chip experiments, we demonstrate that Runx1, together with other key niche factors, is a direct target gene of Gata3. In addition, we can pinpoint Gata3 binding to the Runx1 locus at specific enhancer elements which are active in the microenvironment. These results reveal a direct interaction between Gata3 and Runx1 in the niche that supports embryonic HSCs and highlight a dual role for Runx1 in driving the transdifferentiation of haemogenic endothelial cells into HSCs as well as in the stromal cells that support this process.


Using a new analysis of the best interests standard to address cultural disputes: whose data, which values?
Clinicians sometimes disagree about how much to honor surrogates' deeply held cultural values or traditions when they differ from those of the host country. Such a controversy arose when parents requested a cultural accommodation to let their infant die by withdrawing life saving care. While both the parents and clinicians claimed to be using the Best Interests Standard to decide what to do, they were at an impasse. This standard is analyzed into three necessary and jointly sufficient conditions and used to resolve the question of how much to accommodate cultural preferences and how to treat this infant. The extreme versions of absolutism and relativism are rejected. Properly understood, the Best Interests Standard can serve as a powerful tool in settling disputes about how to make good decisions for those who cannot decide for themselves.


Rehabilitation of locomotion after spinal cord injury.
Advances in our understanding of the control of locomotion enable us to optimize the rehabilitation of patients with a spinal cord injury (SCI). Based on various animal models, it is generally accepted that central pattern generators (CPG) exists for the rhythmic generation of stepping movements, and that this is also the case in humans. However, in humans supraspinal control is also essential for the performance of locomotion. For regaining locomotor function, incomplete SCI subjects strongly depend on visual input to compensate for proprioceptive deficits and impaired balance. In addition, they require additional attentional capacity to stand, walk and handle their walking aids. These factors might contribute to their higher risk of falling. During the last decade, task-specific functional training performed by physiotherapists, combined with manual or robotic assisted bodyweight supported treadmill training have improved the regaining of ambulatory function in patients with incomplete SCI. At present, there is no difference in effectiveness between these three types of training. In the future, rehabilitation programs should be optimized to maximally exploit spontaneous and induced neural plasticity, leading to improved ambulation. To evaluate the efficacy of rehabilitation programs and of experimental treatments that might be translated from bench to bedside within the next few years, several objective assessments such as the 10 meter walk test and Walking Index for Spinal Cord Injury have been successfully introduced in the field of SCI rehabilitation.