The mystery of the split earlobe.
The ancient art of body piercing has rejuvenated in the recent years as part of the fashion process. The ear is the most frequent body part to be pierced to wear jewelry. Split earlobes are commonly presented to plastic surgeons and the recurrence rate is high. The etiology of the acquired split earlobe was thought to be attributable to either trauma or heavy earrings. In this study, the authors explored the cause of the split earlobe and recurrence after surgical repair. Twenty-five patients who were using gold earrings presented with split earlobe and were studied, and the etiology of the condition was analyzed. A questionnaire was completed and the tissue obtained during surgical repair of the split earlobes was submitted for histopathological studies. This group of patients was compared with 17 subjects having stretched earlobe who were using heavy gold earrings. The control group consists of 50 subjects using gold earrings with normal earlobes. Clinical presentation and the histological studies suggest that allergy to metals used in the earring could lead to split earlobe. There is a difference between the split earlobe and stretched earlobe; the latter results from constant pull by heavy earrings. The authors present a new theory regarding the etiology of split earlobe and recommend that avoiding the offending metal in the earring is indispensable to prevent recurrence.


Construction cost and invasive potential: comparing Lythrum salicaria (Lythraceae) with co-occurring native species along pond banks.
Lythrum salicaria (purple loosestrife) is a nonindigenous invasive species characterized by prolific growth and abundance in marshy and riparian habitats across North America. Given its invasive success, we hypothesized this species may require less energy and/or use energy more efficiently for biomass construction than co-occurring noninvasive plant species. We measured leaf construction cost (CC), leaf mass per unit area (LMA), and leaf organic nitrogen and carbon content of L. salicaria and the five most abundant co-occurring species, Parthenocissus quinquefolia, Erigeron philadelphicus, Asclepias syriaca, Spiraea latifolia, and Solidago graminifolia, along dammed ponds in the Black Rock Forest, Cornwall, New York, USA. Lythrum salicaria, which was highly abundant (2.52 individuals/m(2)), exhibited significantly lower area-based leaf CC (44.47 ± 4.24 g glucose/m(2) leaf) than relatively less abundant species, suggesting energetics may influence its invasive success. Conversely, least abundant Solidago graminifolia (0.67 individuals/m(2)) exhibited the significantly highest leaf CC per unit leaf area (141.87 ± 39.21 g glucose/m(2) leaf). Overall, a negative correlation between species abundance and area-based leaf CC (r(2) = 0.73) indicated low energy requirements and/or high energy efficiency may influence relative abundance in the plant species studied. As it correlates with species abundance in this study, CC may be a useful tool for evaluating invasive potential.


Electrochemical treatment of industrial wastewater containing 5-amino-6-methyl-2-benzimidazolone: toward an electrochemical-biological coupling.
We studied the electrochemical oxidation, on Pt anodes, of industrial wastewaters containing 5-amino-6-methyl-2-benzimidazolone (AMBI). Electrolysis of this non-biodegradable effluent produces simultaneous oxidation of AMBI and chloride ions. Highly oxidative chlorine intermediate species further boost the degradation of AMBI. Solution temperature, pH and current density affect little the degradation of AMBI. At our best conditions, AMBI was 100% degraded in 45 min. However, because the reaction intermediates exhibited high toxicity and non-biodegradability, the electrolysis had to continue for 3 more hours in order to obtain a biocompatible solution. Then, complete mineralization of the outputs from the electrolytic cells was readily achieved in a fixed bed biological reactor.


Indigenous uses of wild and tended plant biodiversity maintain ecosystem services in agricultural landscapes of the Terai Plains of Nepal.
Despite a rapidly accumulating evidence base quantifying ecosystem services, the role of biodiversity in the maintenance of ecosystem services in shared human-nature environments is still understudied, as is how indigenous and agriculturally dependent communities perceive, use, and manage biodiversity. The present study aims to document traditional ethnobotanical knowledge of the ecosystem service benefits derived from wild and tended plants in rice-cultivated agroecosystems, compare this to botanical surveys, and analyze the extent to which ecosystem services contribute social-ecological resilience in the Terai Plains of Nepal. Sampling was carried out in four landscapes, 22 Village District Committees, and 40 wards in the monsoon season. Data collection was based on transects walks to collect plant specimens, structured and semi-structured interviews, and participatory fieldwork in and around home gardens, farms, and production landscapes. We asked 180 farmers to free-list vernacular names and describe use-value of wild and tended plants in rice-cultivated agroecosystems. Uses were categorized into eight broad groupings, and 61 biomedical ailment classifications. We assessed if knowledge of plant species diversity and abundance differed with regard to caste, age, and gender. Nepalese farmers have a deep knowledge of the use and management of the 391 vascular plant specimens identified, which provide key provisioning, regulating, supporting, and cultural ecosystem services. Altogether, plants belong to 76 distinct plant species from 49 phylogenetic families: 56 are used to cure 61 ailments, 27 for rituals, 25 for food, 20 for timber, 17 for fuel, 17 for fodder, 11 for soil enhancement, and eight for pesticides. Four caste groups have statistically different knowledge, and younger informants report a lower average number of useful plants. Agricultural landscapes in Nepal are reservoirs of biodiversity. The knowledge of the use of wild and tended plant species in and around these farms differs by the caste and age group of land manager. Conducting research on agroecosystems will contribute to a deeper understanding of how nature is perceived by locals, to more efficient management and conservation of the breadbasket of Nepal, and to the conservation of valuable, but disappearing traditional knowledge and practice.


Protection against perceptions of powerlessness and helplessness during palliative care: the family members' perspective.
Resilience in relation to coping with stress, loss, and bereavement has recently received increased attention. The aim of the current study was to describe aspects that are experienced as a protection against powerlessness and/or helplessness during advanced palliative home care (APHC) or as a help when coping with such perceptions. Both family members during ongoing APHC and family members 3-9 months after the patient's death responded (in total, N = 233; response rate 72%) to a postal questionnaire with mainly open-ended questions. The text responses were analyzed using Manifest Content Analysis. Protection against powerlessness and helplessness had been facilitated by a stable patient condition, the patient coping well, a trusting relationship with the patient, practical and emotional support from family and friends, access to palliative expertise, and staff support that was both individually-focused and cooperative. Other aspects that had helped or protected family members against powerlessness and helplessness were a belief that they had their own reliable knowledge to manage the difficult situation, talking to someone, doing good for the patient, distracting activities, acceptance, meaning and hope, and an inner feeling of security. The findings are discussed in relation to existential psychology, the dual process model of coping with bereavement, and repressive coping. Clinical implications are suggested.


Diabetes and Diet: Purchasing Behavior Change in Response to Health Information.
Individuals with obesity and related conditions are often reluctant to change their diet. Evaluating the details of this reluctance is hampered by limited data. I use household scanner data to estimate food purchase response to a diagnosis of diabetes. I use a machine learning approach to infer diagnosis from purchases of diabetes-related products. On average, households show significant, but relatively small, calorie reductions. These reductions are concentrated in unhealthy foods, suggesting they reflect real efforts to improve diet. There is some heterogeneity in calorie changes across households, although this heterogeneity is not well predicted by demographics or baseline diet, despite large correlations between these factors and diagnosis. I suggest a theory of behavior change which may explain the limited overall change and the fact that heterogeneity is not predictable.


Rapid simultaneous detection of enterovirus and parechovirus RNAs in clinical samples by one-step real-time reverse transcription-PCR assay.
Enteroviruses (EVs) are recognized as the major etiological agent in meningitis in children and young adults. The use of molecular techniques, such as PCR, has substantially improved the sensitivity of enterovirus detection compared to that of virus culture methods. PCR-based methods also can detect a much wider range of EV variants, including those within species A, as well as human parechoviruses (HPeVs) that often grow poorly in vitro and which previously have been underdiagnosed by traditional methods. To exploit these developments, we developed a real-time one-step reverse transcription-PCR (RT-PCR) for the rapid and sensitive detection of EV and HPeV in clinical specimens. Two commercially available RT-PCR kits were used (method I, Platinum one-step kit; method II, Express qPCR one-step kit) with primers and probes targeting the EV and HPeV 5'-untranslated regions (5'UTR). Amplification dynamics (threshold cycle [C(T)]values and efficiencies) of absolutely quantified full-length RNA transcripts representative of EV species A to D and HPeV were similar, demonstrating the effectiveness of both assays across the range of currently described human EV and HPeV variants. Probit analysis of multiple endpoint replicates demonstrated comparable sensitivities of the assays for EV and HPeV (method I, approximately 10 copies per reaction for both targets; method II, 20 copies per reaction). C(T) values were highly reproducible on repeat testing of positive controls within assays and between assay runs. Considering the sample turnaround time of less than 3 h, the multiplexed one-step RT-PCR method provides rapid diagnostic testing for EV and HPeV in cases of suspected central nervous system infections in a clinically relevant time frame.


Occurrence and complications of tracheal reintubation in critically ill adults.
Timing and preparation for tracheal extubation are as critical as the initial intubation. There are limited data on specific strategies for a planned extubation. The extent to which the difficult airway at reintubation contributes to patient morbidity is unknown. The aim of the present study was to describe the occurrence and complications of failed extubation and associated risk factors, and to estimate the mortality and morbidity associated with reintubation attempts. Cohort study of 2,007 critically ill adult patients admitted to the ICU with an ETT. Patients were classified in 2 groups, based on the requirement for reintubation: "never reintubated" versus "≥ 1 reintubations." Baseline characteristics, ICU and hospital stay, hospital mortality, and in-patient costs were compared between patients successfully extubated and those with reintubation outside the operating room, using regression techniques. Reasons, airway management techniques, and complications of intubation and reintubation were summarized descriptively. 376 patients (19%) required reintubation, and 230 (11%) were reintubated within 48 hours, primarily due to respiratory failure. Patients requiring reintubation were older, more likely to be male, and had higher admission severity score. Difficult intubation and complications were similar for initial and subsequent intubation. Reintubation was associated with a 5-fold increase in the relative odds of death (adjusted odds ratio 5.86, 95% CI 3.87-8.89, P < .01), and a 2-fold increase in median ICU and hospital stay, and institutional costs. Difficult airway at reintubation was associated with higher mortality (adjusted odds ratio 2.23, 95% CI 1.01-4.93, P = .05). Nearly 20% of critically ill patients required out of operating room reintubation. Reintubation was associated with higher mortality, stay, and cost. Moreover, a difficult airway at reintubation was associated with higher mortality.


An Enhanced Visualization Method to Aid Behavioral Trajectory Pattern Recognition Infrastructure for Big Longitudinal Data.
Big longitudinal data provide more reliable information for decision making and are common in all kinds of fields. Trajectory pattern recognition is in an urgent need to discover important structures for such data. Developing better and more computationally-efficient visualization tool is crucial to guide this technique. This paper proposes an enhanced projection pursuit (EPP) method to better project and visualize the structures (e.g. clusters) of big high-dimensional (HD) longitudinal data on a lower-dimensional plane. Unlike classic PP methods potentially useful for longitudinal data, EPP is built upon nonlinear mapping algorithms to compute its stress (error) function by balancing the paired weights for between and within structure stress while preserving original structure membership in the high-dimensional space. Specifically, EPP solves an NP hard optimization problem by integrating gradual optimization and non-linear mapping algorithms, and automates the searching of an optimal number of iterations to display a stable structure for varying sample sizes and dimensions. Using publicized UCI and real longitudinal clinical trial datasets as well as simulation, EPP demonstrates its better performance in visualizing big HD longitudinal data.


Resistance exercise as a countermeasure to disuse-induced bone loss.
During spaceflight, skeletal unloading results in loss of bone mineral density (BMD). This occurs primarily in the spine and lower body regions. This loss of skeletal mass could prove hazardous to astronauts on flights of long duration. In this study, intense resistance exercise was used to test whether a training regimen would prevent the loss of BMD that accompanies disuse. Nine subjects (5 men, 4 women) participated in a supine maximal resistance exercise training program during 17 wk of horizontal bed rest. These subjects were compared with 18 control subjects (13 men, 5 women) who followed the same bed rest protocol without exercise. Determination of treatment effect was based on measures of BMD, bone metabolism markers, and calcium balance obtained before, during, and after bed rest. Exercisers and controls had significantly (P < 0.05) different means, represented by the respective following percent changes: lumbar spine BMD, +3% vs. -1%; total hip BMD, +1% vs. -3%; calcaneus BMD, +1% vs. -9%; pelvis BMD, -0.5% vs. -3%; total body BMD, 0% vs. -1%; bone-specific alkaline phosphatase, +64% vs. 0%; alkaline phosphatase, +31% vs. +5%; osteocalcin, +43% vs. +10%; 1,25 dihydroxyvitamin D, +12% vs. -15%; parathyroid hormone intact molecule, +18% vs. -25%; and serum and ionized calcium, -1% vs. +1%. The difference in net calcium balance was also significant (+21 mg/day vs. -199 mg/day, exercise vs. control). The gastrocnemius and soleus muscle volumes decreased significantly in the exercise group, but the loss was significantly less than observed in the control group. The results indicate that resistance exercise had a positive treatment effect and thus might be useful as a countermeasure to prevent the deleterious skeletal changes associated with long-duration spaceflight.


Personalised Medicine Approaches to Screening and Prevention.
One of the promised benefits of 'personalised' medicine is that it will have a positive impact on disease screening and prevention, by enabling more person-specific estimates of risk, and hence more personalised strategies for screening and risk reduction. This article summarises some of the areas in which these approaches are already being implemented, but also outlines some of the potential difficulties and limitations - and challenges the assumption that 'personalised' approaches are necessarily superior for prevention and screening.


A genetic variant BDNF polymorphism alters extinction learning in both mouse and human.
Mouse models are useful for studying genes involved in behavior, but whether they are relevant to human behavior is unclear. Here, we identified parallel phenotypes in mice and humans resulting from a common single-nucleotide polymorphism in the brain-derived neurotrophic factor (BDNF) gene, which is involved in anxiety-related behavior. An inbred genetic knock-in mouse strain expressing the variant BDNF recapitulated the phenotypic effects of the human polymorphism. Both were impaired in extinguishing a conditioned fear response, which was paralleled by atypical frontoamygdala activity in humans. Thus, this variant BDNF allele may play a role in anxiety disorders showing impaired learning of cues that signal safety versus threat and in the efficacy of treatments that rely on extinction mechanisms, such as exposure therapy.


Tb3+-Cleavage Assays Reveal Specific Mg2+ Binding Sites Necessary to Pre-fold the btuB Riboswitch for AdoCbl Binding.
Riboswitches are RNA elements that bind specific metabolites in order to regulate the gene expression involved in controlling the cellular concentration of the respective molecule or ion. Ligand recognition is mostly facilitated by Mg2+ mediated pre-organization of the riboswitch to an active tertiary fold. To predict these specific Mg2+ induced tertiary interactions of the btuB riboswitch from E. coli, we here report Mg2+ binding pockets in its aptameric part in both, the ligand-free and the ligand-bound form. An ensemble of weak and strong metal ion binding sites distributed over the entire aptamer was detected by terbium(III) cleavage assays, Tb3+ being an established Mg2+ mimic. Interestingly many of the Mn+ (n = 2 or 3) binding sites involve conserved bases within the class of coenzyme B12-binding riboswitches. Comparison with the published crystal structure of the coenzyme B12 riboswitch of S. thermophilum aided in identifying a common set of Mn+ binding sites that might be crucial for tertiary interactions involved in the organization of the aptamer. Our results suggest that Mn+ binding at strategic locations of the btuB riboswitch indeed facilitates the assembly of the binding pocket needed for ligand recognition. Binding of the specific ligand, coenzyme B12 (AdoCbl), to the btuB aptamer does however not lead to drastic alterations of these Mn+ binding cores, indicating the lack of a major rearrangement within the three-dimensional structure of the RNA. This finding is strengthened by Tb3+ mediated footprints of the riboswitch's structure in its ligand-free and ligand-bound state indicating that AdoCbl indeed induces local changes rather than a global structural rearrangement.


Modified Atkins diet is an effective treatment for children with Doose syndrome.
Children with myoclonic astatic epilepsy (MAE; Doose syndrome) whose seizures do not respond immediately to standard antiepileptic drugs (AEDs) are at high risk of developing an epileptic encephalopathy with cognitive decline. A classic ketogenic diet (KD) is a highly effective alternative to AEDs. To date, there are only limited data on the effectiveness of the modified Atkins diet (MAD), which is less restrictive and more compatible with daily life. We report findings from a retrospective study on 30 MAE patients treated with MAD. Four participating centers retrospectively identified all patients with MAE in whom a MAD had been started before June 2015. Seven children were recruited from a cohort included in an open prospective controlled trial. A retrospective review of all available charts was performed in the other patients. Thirty patients (24 boys) were included. Mean age at epilepsy onset was 3.1 years (range 1.5-5.6). MAD was started at a mean age of 4.5 years (range 2.2-9.1) after the children had received an average of six different AEDs (range 2-15). Mean MAD observation time was 18.7 months (range 1.5-61.5). Twenty of 30 patients were still on MAD at the end of study (duration range 1.5-61.5, mean 18.5 months). MAD was stopped without relapse in three patients after sustained seizure freedom for >2 years. For the other seven cases, ineffectiveness (three patients), loss of efficacy (two), or noncompliance (two) led to termination. No severe adverse effects were noted. By the end of the observation period, 25 (83%) of 30 patients experienced a seizure reduction by ≥50% and 14 (47%) of 30 were seizure-free. None of the evaluated factors differed significantly between the groups of seizure-free and non-seizure-free children. MAD is an effective treatment for MAE. It should be considered as an alternative to AEDs or the more restrictive classic ketogenic diet.


[C-reactive protein as a marker of infection in women with preterm delivery].
There is a strong association between systemic and intrauterine infection and preterm delivery. C-reactive protein (CRP) is considered a nonspecific marker for intrauterine inflammation. We determined its blood level in 100 women who presented with preterm labor and intact membranes and delivered prematurely. CRP levels were used as a noninvasive marker for infection to compare clinical characteristics between women who delivered prematurely at 31-36 and 24-30 weeks of gestation. Women who delivered earlier than the 24th week of gestation had a higher rate of positive CRP levels than those who delivered later, 54% vs 24%, respectively (p < 0.0001). Moreover, women from the lower gestational age group with positive CRP levels had significantly different clinical characteristics than those in the same group but with negative CRP levels. There was a significant difference between Jewish and Bedouin women in cervical dilatation and time interval from hospitalization to delivery between those with positive CRP and those with negative CRP levels. We conclude that patients who delivered prematurely at 24-30 weeks had higher rates of an inflammatory etiology than women who also delivered prematurely, but at a more advanced gestational age.


Short-Term Alcohol Abstinence Improves Antibacterial Defenses of Chronic Alcohol-Consuming Mice against Gut Bacteria-Associated Sepsis Caused by Enterococcus faecalis Oral Infection.
The effects of short-term alcohol abstinence on host antibacterial resistance against Enterococcus faecalis oral infection was investigated in chronic alcohol-consuming mice [mice with 0.1 g/day of 20% ethanol consumption for 12 or 16 weeks (CAC-mice)]. These mice were highly susceptible to the infection; however, after 7 days of alcohol abstinence (aaCAC-mice), their antibacterial resistances were completely restored to the normal mouse level. Normal mice inoculated with CAC-mouse hepatic macrophages were shown to be susceptible to the infection, whereas the same macrophage preparation from aaCAC-mice did not impair the antibacterial resistance of normal mice. aaCAC-mouse liver macrophages protected nonobese diabetic-severe combined immunodeficiency IL-2Rγnull mice exposed to E. faecalis, whereas those from CAC-mice did not. Monocyte-derived (MD) M2b macrophages were predominantly isolated from CAC-mouse livers, but these cells were not significantly isolated from aaCAC-mouse livers. Hepatic MD macrophages from aaCAC-mice switched to M1 macrophages in response to bacterial antigen, whereas the same macrophage preparation from CAC-mice did not. M1 Kupffer cells, M2a Kupffer cells, and MD M2b macrophages were shown to be not bactericidal, whereas E. faecalis was killed effectively by M1 macrophages derived from aaCAC-mouse hepatic MD macrophages. These results indicate that MD M2b macrophages predominantly distributed in the liver are responsible for the impaired resistance of CAC-mice to E. faecalis oral infection, and aaCAC-mice without MD M2b macrophages in the livers are resistant to the infection.


Effect of organic matter on estuarine flocculation: a laboratory study using montmorillonite, humic acid, xanthan gum, guar gum and natural estuarine flocs.
Riverine particles undergo a rapid transformation when they reach estuaries. The rapid succession of hydrodynamic and biogeochemical regimes forces the particles to flocculate, settle and enter the sediment pool. The rates and magnitudes of flocculation depend on the nature of the particles which are primarily affected by the types and quantities of organic matter (OM). Meanwhile, the OM characteristics vary widely between environments, as well as within a single environment due to seasonal climate and land use variability. We investigated the effect of the OM types and quantities through laboratory experiments using natural estuarine particles from the Mississippi Sound and Atchafalaya Bay as well as model mixtures of montmorillonite and organic molecules (i.e., biopolymers (guar/xanthan gums) and humic acid). Biopolymers promote flocculation but the magnitude depends on the types and quantities. Nonionic guar gum yields much larger flocs than anionic xanthan gum, while both of them exhibit a nonlinear behavior in which the flocculation is the most pronounced at the intermediate OM loading. Moreover, the effect of guar gum is independent of salinity whereas the effect of xanthan gum is pronounced at higher salinity. Meanwhile, humic acid does not affect flocculation at all salinity values tested in this study. These results are echoed in the laboratory manipulation of the natural estuarine particles. Flocculation of the humic acid-rich Mississippi Sound particles is unaffected by the OM, whereas that of biopolymer-rich Atchafalaya Bay particles is enhanced by the OM. Flocculation is positively influenced by the presence of biopolymers that are produced as the result of marine primary production. Meanwhile, humic acid, which is abundant in the rivers that drain the agricultural soils of Southeastern United States, has little influence on flocculation. Thus, it is expected that humic acid-poor riverine particles (e.g., Mississippi River, and Atchafalaya River, to a lesser degree) may be prone to rapid flocculation and settling in the immediate vicinity of the river mouths when mixed with biopolymer-rich coastal waters. It is also expected that humic acid-rich riverine particles (e.g., Pearl River) may resist immediate flocculation and be transported further away from the river mouth.


An update on therapeutic angiogenesis for peripheral vascular disease.
We reviewed the issue of stem cells and therapeutic angiogenesis in the treatment of peripheral vascular disease. MEDLINE (1997-2008) with the following search terms: "stem cell therapy," "endothelial progenitor cells," "peripheral blood mononuclear cells," and "peripheral vascular disease." Relevant published papers involving the above search terms, preclinical studies, and clinical trials using stem cells and progenitors for the treatment of peripheral occlusive vascular disease were included. Transplantation of bone marrow-derived progenitor cells or peripheral blood mononuclear cells promotes tissue angiogenesis, as has already been explored in preclinical studies; angiogenesis can also be sustained using genetic, protein therapeutic approaches. Engineered scaffolding with stem cells is a further strategy, which is still in its infancy. The treatment of patients with severe peripheral arterial disease is generally reported as a series of case reports; all studies generally show an improvement in clinical symptoms, e.g., rest pain and pain-free walking time, as well as transcutaneous oxygen pressure, without any important adverse reactions. The few clinical trials also report similar encouraging results. All the studies have their shortcomings, including absence of control groups and objective evaluation of the results of treatments as well as short-term follow-up. Promoting angiogenesis using genetic, protein, stem cell-based therapies is a promising option for the treatment of peripheral vascular disease when unresponsive to medical and surgical therapy.


Vision modulation, plasticity and restoration using non-invasive brain stimulation - An IFCN-sponsored review.
The visual system has one of the most complex structures of all sensory systems and is perhaps the most important sense for everyday life. Its functional organization was extensively studied for decades in animal and humans, for example by correlating circumscribed anatomical lesions in patients with the resulting visual dysfunction. During the past two decades, significant achievements were accomplished in characterizing and modulating visual information processing using non-invasive stimulation techniques of the normal and damaged human eye and brain. Techniques include transcranial magnetic stimulation (TMS) and low intensity electric stimulation using either direct or alternating currents applied transcranially (tDCS or tACS) near or above the visual cortex, or alternating currents applied transorbitally (trACS). In the case of transorbital stimulation of the visual system the electrodes are attached near the eye, to the eyelids (transpalpebral electrical stimulation - TPES) or the cornea (tanscorneal electrical stimulation TcES). Here, we summarize the state-of-the-art of visual system magnetic and electric stimulation as a method to modulate normal vision, induce brain plasticity, and to restore visual functions in patients. We review this field's history, models of current flow paths in the eye and brain, neurophysiological principles (e.g. entrainment and after-effects), the effects on vision in normal subjects and the clinical impact on plasticity and vision restoration in patients with low vision, with a particular focus on "off-line" or "after-effects". With regard to the therapeutic possibilities, ACS was demonstrated to be effective in patients affected by glaucoma and optic neuropathy, while tDCS and random noise stimulation (tRNS) are most promising for the treatment of amblyopia, hemianopia and myopia. In addition, rTMS applied above the occipital area is a promising approach to treat migraine, neglect and hemianopia. Although the response to these treatment options is better than to sham stimulation in double blinded clinical studies, the clinical efficacy is still rather variable and a proportion of patients do not respond. It is therefore imperative to better understand the mechanisms of action to be able to optimize treatment protocols possibly through personalization of brain stimulation protocols. By identifying the current opportunities and challenges in the field, we hope to provide insights to help improve neuromodulation protocols to restore visual function in patients with visual system damage.


Motivation-dependent responses in the human caudate nucleus.
Motivation is a complex process that leads to completion or avoidance of a behavior. Past research strongly implicates the basal ganglia in a circuit integral for the control of motivation. Specifically, the human striatum has been shown to process reward information, differentiating between monetary rewards and punishments in recent neuroimaging experiments. It is unclear, however, how the dorsal striatum, particularly the caudate nucleus, responds to changes in the motivational context of a task. Using an event-related design, where participants were given positive and negative feedback upon guessing the value of an unknown card, we manipulated the motivational context of the task by dividing trials into periods of high incentive (where visual feedback indicated monetary rewards and punishments) and low incentive (where visual feedback indicated only accuracy). We found that activity in the caudate nucleus was strongly influenced by the different incentive periods. The hemodynamic response was characterized by a larger rise at the onset of trials and larger differences between positive and negative feedback during periods of high incentive. These results suggest that changes in motivation are capable of modulating basal ganglia activity, and further support an important role for the caudate nucleus in affective processing.


Restaging Patients With Hepatocellular Carcinoma Before Additional Treatment Decisions: A Multicenter Cohort Study.
Prognostic assessment of patients with hepatocellular carcinoma (HCC) at the time of diagnosis remains controversial and becomes even more complex at the time of restaging when new variables need to be considered. The aim of the current study was to evaluate the prognostic utility of restaging patients before proceeding with additional therapies for HCC. Two independent Italian prospective databases were used to identify 1,196 (training cohort) and 648 (validation cohort) consecutive patients with HCC treated over the same study period (2008-2015) who had complete restaging before decisions about additional therapies. The performance of the Italian Liver Cancer (ITA.LI.CA) prognostic score at restaging was compared with that of the Barcelona Clinic Liver Cancer, Hong Kong Liver Cancer, and Cancer of the Liver Italian Program systems. A multivariable Cox survival analysis was performed to identify baseline, restaging, or dynamic variables that were able to improve the predictive performance of the prognostic systems. At restaging, 35.3% of patients maintained stable disease; most patients were either down-staged by treatment (27.2%) or had disease progression (37.5%). The ITA.LI.CA scoring system at restaging demonstrated the best prognostic performance in both the training and validation cohorts (c-index 0.707 and 0.722, respectively) among all systems examined. On multivariable analysis, several variables improved the prognostic ability of the ITA.LI.CA score at restaging, including progressive disease after the first treatment, Model for End-Stage Liver Disease at restaging, and choice of nonsurgical treatment as additional therapy. A new ITA.LI.CA restaging model was created that demonstrated high discriminative power in both the training and validation cohorts (c-index 0.753 and 0.745, respectively). Although the ITA.LI.CA score demonstrated the best prognostic performance at restaging, other variables should be considered to improve the prognostic assessment of patients at the time of deciding additional therapies for HCC.


In situ FTIR spectroscopy study of the photodegradation of acetaldehyde and azo dye photobleaching on bismuth-modified TiO2.
The photocatalytic properties of bismuth-modified titania were studied by photobleaching of two aqueous azo dyes solutions (Reactive Black 5 and Acid Orange 7), and by photoinduced decomposition (PID) of acetaldehyde using in situ FTIR spectroscopy. Low bismuth doping concentrations up to 3 at.% is shown to lead to an increased photobleaching rate of both azo dyes solutions. Too high Bi dopant concentrations lead to less developed crystallite nanoparticles and exhibit weaker adsorption capacity. Bismuth doping altered the adsorption kinetics of acetaldehyde resulting in different surface products, and a modified photocatalytic reaction pathway was inferred.


Breast carcinoma and phyllodes tumour: a case series.
Malignant transformation of the epithelial component of phyllodes tumours (PT) is rare and only reported in literature as sporadic cases of carcinoma associated with PTs. We report the clinicopathological characteristics of in situ and invasive carcinoma coexisting with PT in 10 patients treated in our institution over an 11-year period from 1992 to 2012. Ten patients with coexisting PT and in situ or invasive carcinoma were identified from our records. Six had carcinoma found within the PT. All were female with a median age of 47 (43-72) years. One patient had a history of PT in the same breast while another had a history of PT in the same breast as well as invasive ductal carcinoma in the contralateral breast. The rest did not have any risk factors of breast cancer. Five patients had a preoperative core needle biopsy performed with the report of a fibroepithelial lesion. The rest of the patients had surgery upfront for their breast masses. Two patients who had ER/PR positive invasive carcinoma received adjuvant hormonal therapy. Patients were followed up for a mean of 3.6 years (9 months-10 years) and all patients were alive and recurrence free. PT associated with carcinoma is rare, and we present a series of cases that add to the limited current literature. It is often difficult to detect the presence of the carcinomatous component preoperatively. Hence, close examination of resected PT specimens must be carried out to allow prompt detection of any associated carcinomas, however rare, such that adequate treatment can be given.


From causes to solutions--insights from lay knowledge about health inequalities.
This paper reports on a qualitative study of lay knowledge about health inequalities and solutions to address them. Social determinants of health are responsible for a large proportion of health inequalities (unequal levels of health status) and inequities (unfair access to health services and resources) within and between countries. Despite an expanding evidence base supporting action on social determinants, understanding of the impact of these determinants is not widespread and political will appears to be lacking. A small but growing body of research has explored how ordinary people theorise health inequalities and the implications for taking action. The findings are variable, however, in terms of an emphasis on structure versus individual agency and the relationship between being 'at risk' and acceptance of social/structural explanations. This paper draws on findings from a qualitative study conducted in Adelaide, South Australia, to examine these questions. The study was an integral part of mixed-methods research on the links between urban location, social capital and health. It comprised 80 in-depth interviews with residents in four locations with contrasting socio-economic status. The respondents were asked about the cause of inequalities and actions that could be taken by governments to address them. Although generally willing to discuss health inequalities, many study participants tended to explain the latter in terms of individual behaviours and attitudes rather than social/structural conditions. Moreover, those who identified social/structural causes tended to emphasise individualized factors when describing typical pathways to health outcomes. This pattern appeared largely independent of participants' own experience of advantage or disadvantage, and was reinforced in discussion of strategies to address health inequalities. Despite the explicit emphasis on social/structural issues expressed in the study focus and framing of the research questions, participants did not display a high level of knowledge about the nature and causes of place-based health inequalities. By extending the scope of lay theorizing to include a focus on solutions, this study offers additional insights for public health. Specifically it suggests that a popular constituency for action on the social determinants of health is unlikely to eventuate from the current popular understandings of possible policy levers.


Seroprevalence of human enterovirus 71 and coxsackievirus A16 in Guangdong, China, in pre- and post-2010 HFMD epidemic period.
Human Enterovirus 71 and Coxsackie A16 have caused many outbreaks in the last decade in mainland China, resulting in thousands of fatal cases. Seroepidemiology which provides important information to document population immunity is rare in China. A cross sectional study of Enterovirus 71 (EV71) and Coxsackie A16 (CA16) seroprevalence was carried out in Guangdong, China, pre- and post- the 2010 hand, foot and mouth disease (HFMD) epidemic period. The levels of EV71 and CA16 specific antibodies were evaluated by a microneutralization test and the geometric mean titer (GMT) was calculated and compared. Our results indicated frequent infection by EV71 and CA16 in Guangdong before the 2010 epidemic. Only EV71 neutralizing antibody but not CA16 seroprevalence was significantly increased after the 2010 HFMD epidemic. Children less than 3 years old especially those aged 2 years showed the lowest positive rates for EV71 and CA16 NA before epidemic and the most significantly increased EV71 seroprevalence after epidemic. CA16 GMT values declined after the 2010 epidemic. These results indicate EV71 was the major pathogen of HFMD in Guangdong during the 2010 epidemic. The infection occurs largely in children less than 3 years, who should have first priority to receive an EV71 vaccine.


Systematic Comparison of High-throughput Single-Cell and Single-Nucleus Transcriptomes during Cardiomyocyte Differentiation.
A comprehensive reference map of all cell types in the human body is necessary for improving our understanding of fundamental biological processes and in diagnosing and treating disease. High-throughput single-cell RNA sequencing techniques have emerged as powerful tools to identify and characterize cell types in complex and heterogeneous tissues. However, extracting intact cells from tissues and organs is often technically challenging or impossible, for example in heart or brain tissue. Single-nucleus RNA sequencing provides an alternative way to obtain transcriptome profiles of such tissues. To systematically assess the differences between high-throughput single-cell and single-nuclei RNA-seq approaches, we compared Drop-seq and DroNc-seq, two microfluidic-based 3' RNA capture technologies that profile total cellular and nuclear RNA, respectively, during a time course experiment of human induced pluripotent stem cells (iPSCs) differentiating into cardiomyocytes. Clustering of time-series transcriptomes from Drop-seq and DroNc-seq revealed six distinct cell types, five of which were found in both techniques. Furthermore, single-cell trajectories reconstructed from both techniques reproduced expected differentiation dynamics. We then applied DroNc-seq to postmortem heart tissue to test its performance on heterogeneous human tissue samples. Our data confirm that DroNc-seq yields similar results to Drop-seq on matched samples and can be successfully used to generate reference maps for the human cell atlas.


Impaired leptin responsiveness in aged rats.
We previously reported that adiposity and serum leptin levels increase with age in male F-344xBN rats and that when physiological levels of serum leptin are manipulated by fasting, there is a corresponding reciprocal change in hypothalamic neuropeptide Y (NPY) mRNA in young rats, but there are no changes in older rats. These findings suggest that the regulation of hypothalamic NPY mRNA by leptin may be impaired with age. To test this hypothesis, we infused saline or leptin for 7 days into ad libitum-fed rats and compared these with saline-infused rats that were pair-fed the amount of food consumed by the leptin-treated rats. We examined daily food consumption, body weight, whole-body oxygen consumption, serum leptin, and NPY mRNA in the hypothalamus. Food consumption decreased by 50% in the leptin-infused compared with the saline-infused young rats but only decreased by 20% in the aged rats. In the leptin-treated young rats, there was a 24% increase in oxygen consumption compared with the pair-fed rats, but there were no changes in oxygen consumption in the aged rats. Leptin infusion diminished hypothalamic NPY levels by nearly 50% compared with pair-fed young rats, whereas there were no changes in the hypothalamic NPY mRNA levels in senescent rats. In summary, aged rats demonstrate a reduced responsiveness to leptin, including a diminished decrease in food intake and no increase in energy expenditure. These diminished responses to leptin were associated with and may be the result of an impaired suppression of hypothalamic NPY mRNA levels. This leptin resistance may be due to either the elevated obesity and serum leptin with age or due to age itself, or both.


Neonatal sludge: a finding of congenital hypothyroidism.
Congenital hypothyroidism is one of the most urgent diseases of the neonate. When diagnosed and treated at an early stage, its most important complication, mental retardation, is preventable. The signs of congenital hypothyroidism are nonspecific in neonates. Only 5% of the cases have characteristic clinical findings. One of the most important and earliest signs is prolonged jaundice during the neonatal period. We report herein a case of congenital hypothyroidism, who presented with icterus accompanied with sludge formation into the gallbladder, which disappeared after treatment with L-thyroxine.


In Vitro Metabolism of Isopropylated and tert-Butylated Triarylphosphate Esters Using Human Liver Subcellular Fractions.
Isopropylated and tert-butylated triarylphosphate esters (ITPs and TBPPs, respectively) are plasticizers and flame retardants that are ubiquitous in indoor environments; however, no studies to date have characterized their metabolism. Using human liver subcellular S9 fractions, phase I and II in vitro metabolism of triphenyl phosphate (TPHP), 4-tert-butylphenyl diphenyl phosphate (4tBPDPP), 2-isopropylphenyl diphenyl phosphate (2IPPDPP), and 4-isopropylphenyl diphenyl phosphate (4IPPDPP) was investigated at 1 and 10 μM doses. Parent depletion and the formation of known or suspected metabolites (e.g., likely hydrolysis or hydroxylated products), including diphenyl phosphate (DPHP), hydroxyl-triphenyl phosphate (OH-TPHP), isopropylphenyl phenyl phosphate (ip-PPP), and tert-butylphenyl phenyl phosphate (tb-PPP), were monitored and quantified via GC/MS or LC-MS/MS. tb-PPP and its conjugates were identified as the major in vitro metabolites of 4tBPDPP and accounted for 71% and 49%, respectively, of the parent molecule that was metabolized during the incubation. While the mass balance between parents and metabolites was conserved for TPHP and 4tBPDPP, approximately 20% of the initial parent mass was unaccounted for after quantifying suspected metabolites of 2IPPDPP and 4IPPDPP that had authentic standards available. Two novel ITP metabolites, mono-isopropenylphenyl diphenyl phosphate and hydroxy-isopropylphenyl diphenyl phosphate, were tentatively identified by high-resolution mass spectrometry and screened for in recently collected human urine where mono-isopropenylphenyl diphenyl phosphate was detected in one of nine samples analyzed. This study provides insight into the biological fate of ITP and TBPP isomers in human tissues and is useful in identifying appropriate biomarkers of exposure to monitor, particularly in support of epidemiological studies.


Renal dysfunction and the associated decrease in survival after elective endovascular aneurysm repair.
The reported frequency of renal dysfunction after elective endovascular aneurysm repair (EVAR) varies widely in current surgical literature. Published research establishes pre-existing end-stage renal disease as a poor prognostic indicator. We intend to quantify the mortality effect associated with renal morbidity developed postoperatively and to identify modifiable risk factors. All elective EVAR patients with preoperative and postoperative renal function data captured by the Vascular Quality Initiative between January 2003 and December 2014 were examined. The primary study end point was long-term mortality. Preoperative, intraoperative, and postoperative parameters were analyzed to estimate mortality stratified by renal outcome and to describe independent risk factors associated with post-EVAR renal dysfunction. This study included 14,475 elective EVAR patients, of whom 96.8% developed no post-EVAR renal dysfunction, 2.9% developed acute kidney injury, and 0.4% developed a new hemodialysis requirement. Estimated 5-year survival was significantly different between groups, 77.5% vs 53.5%, respectively, for the no dysfunction and acute kidney injury groups, whereas the new hemodialysis group demonstrated 22.8% 3-year estimated survival (P < .05). New-onset postoperative congestive heart failure (odds ratio [OR], 3.50; 95% confidence interval [CI], 1.18-10.38), return to the operating room (OR, 3.26; 95% CI, 1.49-7.13), and postoperative vasopressor requirement (OR, 2.68; 95% CI, 1.40-5.12) predicted post-EVAR renal dysfunction, whereas a preoperative estimated glomerular filtration rate (eGFR) ≥60 mL/min/1.73 m2 was protective (OR, 0.33; 95% CI, 0.21-0.53). Volume of contrast material administered during elective EVAR varies 10-fold among surgeons in the Vascular Quality Initiative database, but the average volume administered to patients is statistically similar, regardless of preoperative eGFR. Multivariable logistic regression demonstrated nonsignificant correlation between contrast material volume and postoperative renal dysfunction. Any renal dysfunction developing after elective EVAR is associated with decreased estimated long-term survival. Protecting renal function with a rational dosing metric for contrast material linked to preoperative eGFR may better guide treatment.


Fermentation of mucin and plant polysaccharides by strains of Bacteroides from the human colon.
Ten Bacteroides species found in the human colon were surveyed for their ability to ferment mucins and plant polysaccharides ("dietary fiber"). A number of strains fermented mucopolysaccharides (heparin, hyaluronate, and chondroitin sulfate) and ovomucoid. Only 3 of the 188 strains tested fermented beef submaxillary mucin, and none fermented porcine gastric mucin. Many of the Bacteroides strains tested were also able to ferment a variety of plant polysaccharides, including amylose, dextran, pectin, gum tragacanth, gum guar, larch arabinogalactan, alginate, and laminarin. Some plant polysaccharides such as gum arabic, gum karaya, gum ghatti and fucoidan, were not utilized by any of the strains tested. The ability to utilize mucins and plant polysaccharides varied considerably among the Bacteroides species tested.


Transient elastography for diagnosis of stages of hepatic fibrosis and cirrhosis in people with alcoholic liver disease.
The presence and progression of hepatic (liver) fibrosis into cirrhosis is a prognostic variable having impact on survival in people with alcoholic liver disease. Liver biopsy, although an invasive method, is the recommended 'reference standard' for diagnosis and staging of hepatic fibrosis in people with liver diseases. Transient elastography is a non-invasive method for assessing and staging hepatic fibrosis. To determine the diagnostic accuracy of transient elastography for diagnosis and staging hepatic fibrosis in people with alcoholic liver disease when compared with liver biopsy. To identify the optimal cut-off values for differentiating the five stages of hepatic fibrosis. The Cochrane Hepato-Biliary Group Controlled and Diagnostic Test Accuracy Studies Registers, The Cochrane Library, MEDLINE (OvidSP), EMBASE (OvidSP), and the Science Citation Index Expanded (last search August 2014). Diagnostic cohort and diagnostic case-control study designs that assessed hepatic fibrosis in participants with alcoholic liver disease with transient elastography and liver biopsy, irrespective of language or publication status. The study participants could be of any sex and ethnic origin, above 16 years old, hospitalised or managed as outpatients. We excluded participants with viral hepatitis, autoimmunity, metabolic diseases, and toxins. We followed the guidelines in the draft Cochrane Handbook for Systematic Reviews of Diagnostic Test Accuracy. Five retrospective and nine prospective cohort studies with 834 participants provided data for the review analyses. Authors of seven of those studies sent us individual participant data. The risk of bias in the included studies was high in all but three studies. We could identify no serious concerns regarding the applicability of the studies in answering the main study question of our review, namely to use transient elastography to diagnose hepatic fibrosis. We could not identify the optimal cut-off values for the fibrosis stages. The definition of the diagnosis of alcoholic liver disease was not provided in one study and was not clearly defined in two studies, but it was clear in the remaining 11 studies. The study authors used different liver stiffness cut-off values of transient elastography for the hepatic fibrosis stages.There was only one study (103 participants) with data on hepatic fibrosis stage F1 or worse, with a cut-off of 5.9 kPa, and reporting sensitivity of 0.83 (95% confidence interval (CI) 0.74 to 0.90) and specificity of 0.88 (95% CI 0.47 to 1.00). The summary sensitivity and specificity of transient elastography for F2 or worse (seven studies with 338 participants and with cut-offs around 7.5 kPa (range 7.00 to 7.8 kPa)) were 0.94 and 0.89 with LR+ 8.2 and LR- 0.07, which suggests that transient elastography could be useful to rule out the presence of significant hepatic fibrosis, thus avoiding liver biopsy.Due to the wide range of cut-off values (from 8.0 to 17.0 kPa) found in the 10 studies with 760 participants with hepatic fibrosis F3 or worse, we fitted a hierarchical summary receiver operating characteristic (HSROC) model and estimated a summary ROC (SROC) curve. The sensitivity of the 10 studies varied from 72% to 100% and the specificity from 59% to 89%. We performed an additional analysis by including the studies with a cut-off value of around and equal to 9.5 kPa (range 8.0 to 11.0 kPa). The summary sensitivity and specificity of transient elastography (eight studies with 564 participants) were 0.92 and 0.70 with LR+ 3.1 and LR- 0.11, which suggests that transient elastography could also be useful to rule out the presence of severe hepatic fibrosis (F3 or worse), avoiding liver biopsy. We carried out a sensitivity analysis by considering only the studies with a cut-off value equal to 9.5 kPa and the result did not differ.We performed an HSROC analysis and reported an SROC curve for hepatic fibrosis stage F4 (cirrhosis). The HSROC analysis suggested that when the cut-off value changes, there is a wide variation in specificity and a more limited variation in sensitivity. We performed an additional analysis with the studies with the most commonly used cut-off value of 12.5 kPa. The summary sensitivity and specificity of transient elastography (seven studies with 330 participants) were 0.95 and 0.71 with LR+ 3.3 and LR- 0.07, which again suggests that transient elastography could be useful to rule out the presence of cirrhosis, avoiding liver biopsy. We identified a small number of studies with a few participants and were unable to include several studies, which raises the risk of outcome reporting bias. With these caveats in mind, transient elastography may be used as a diagnostic method to rule out liver cirrhosis (F4) in people with alcoholic liver disease when the pre-test probability is about 51% (range 15% to 79%). Transient elastography may also help in ruling out severe fibrosis (F3 or worse). Liver biopsy investigation remains an option if the certainty to rule in or rule out the stage of hepatic fibrosis or cirrhosis remains insufficient after a clinical follow-up or any other non-invasive test considered useful by the clinician.The proposed cut-off values for the different stages of hepatic fibrosis may be used in clinical practice, but caution is needed, as those values reported in this review are only the most common cut-off values used by the study authors. The best cut-off values for hepatic fibrosis in people with alcoholic liver disease could not be established yet.In order to diagnose correctly the stage of hepatic fibrosis in people with alcoholic liver disease using transient elastography assessment, the studies should consider a single aetiology. Hepatic fibrosis should be diagnosed with both transient elastography and liver biopsy and in this sequence, and transient elastography cut-off values should be pre-specified and validated. The time interval between the two investigations should not exceed three months, which is the interval mainly valid for people without cirrhosis, and assessment of results should be properly blinded. Only studies with low risk of bias, fulfilling the Standards for Reporting of Diagnostic Accuracy may answer the review question.


Design, synthesis and biological evaluation of novel Schiff base-bridged tetrahydroprotoberberine triazoles as a new type of potential antimicrobial agents.
A series of novel Schiff base-bridged tetrahydroprotoberberine (THPB) triazoles were designed, synthesized and characterized for the first time. Antimicrobial assay showed that some of the prepared compounds exerted stronger antibacterial and antifungal activities than the reference drugs. Especially, THPB triazole 7a gave low MIC values of 0.5, 1 and 2 μg mL-1 against B. yeast, M. luteus and MRSA, respectively. Further experiments indicated that the highly active molecule 7a was able to rapidly kill the MRSA strain and did not trigger the development of bacterial resistance even after 14 passages. The preliminary exploration for the antimicrobial mechanism revealed that compound 7a could effectively intercalate into calf thymus DNA to form a 7a-DNA supramolecular complex, and its Zn2+ complex had the ability to directly cleave pUC19 DNA, which suggested that compound 7a might be a potentially dual-targeting antibacterial molecule. It was also found that compound 7a could be efficiently stored and carried by human serum albumin (HSA), and the hydrophobic interactions and hydrogen bonds played important roles in the transportation of HSA to the active molecule 7a.


Management of anaemia and blood in patients having neck dissections or free flaps for head and neck cancer.
Our main aims were to assess haemoglobin (Hb) concentrations from preoperative assessment to discharge from hospital, and to review which patients had blood transfusions and compliance with national transfusion guidelines. We studied a consecutive series of 131 patients between October 2016 and September 2017 who had either neck dissection or resection and free microvascular tissue transfer. Half the patients had soft tissue free flaps (n = 65), 26% had composite free flaps (n = 34), and 24% neck dissection only (n = 32). Using the WHO definition of anaemia, 4% (1/28) of patients who had neck dissections and 19% (16/85) of those who had free flaps were anaemic preoperatively. The median (IQR) Hb at discharge was 131 (119-144) g/L for patients who had neck dissections, 103 (95-114) g/L for those who had soft free flaps, and 95 (90-104) g/L for those who had composite free flaps. No patients who had neck dissection were given a red blood cell (RBC) transfusion, whereas they were given to 26/99 (26%) of those who had free flaps. Hb concentrations were checked after each unit in 31/39 transfusions (79%). Concentrations for those who had free flaps fell by about 30 g/L from admission to operation, and only four patients were given tranexamic acid peroperatively. Postoperatively Hb remained at similar concentrations until discharge, with 23/98 (24%) given iron orally on discharge. In terms of compliance with blood transfusion guidelines there was a notable absence of the use of tranexamic acid and of iron intravenously. An increase in their use could potentially reduce the number of blood transfusions required and the postoperative incidence of anaemia, and have a favourable effect on outcomes such as complications, fatigue, and overall quality of life.


Quantification of small molecules in plasma with direct analysis in real time tandem mass spectrometry, without sample preparation and liquid chromatographic separation.
Recently, a new ion source, Direct Analysis in Real Time (DART), has been introduced which allows direct biological sample introduction into a mass spectrometry (MS) system. The elimination of conventionally required sample preparation and separation by high-performance liquid chromatography (HPLC) prior to MS analysis represents a remarkable opportunity to reduce assay turn-around time, environmental impact and capital/manpower investment. This new technology initially was used in various qualitative applications to directly detect chemicals on solid surfaces, in liquids and gases. In this study, a DART source operating under ambient pressure with ground potential was installed onto a Sciex 4000 tandem mass spectrometer and employed in the sample analysis of plasma based on direct introduction into the DART-MS/MS system. Reasonable precision and accuracy (%CV and %Error, both <10%) were achieved of a significant number of compounds tested in biological fluids. In addition, the limit of detection for 80% of the tested compounds reached 5 ng/mL or lower which is sufficient for pharmaceutical drug discovery support. Finally, experimental conditions that significantly impacted assay performance were investigated with respect to optimization and limitation. Because of its simplicity, fast data acquisition (3-5 s) and low cost, DART has the potential to significantly impact quantitative pharmaceutical analysis in biological matrices.


Determinants of End-of-Life Expenditures in Patients with Oral Cancer in Taiwan: A Population-Based Study.
To investigate the association of basic demographic data, socioeconomic status, medical services, and hospital characteristics with end-of-life expenditure in patients with oral cancer in Taiwan who died between 2009 to 2011. This nationwide population-based, retrospective cohort study identified 5,386 patients who died from oral cancer. We evaluated medical cost in the last month of life by universal health insurance. The impact of each variable on the end-of-life expenditure was examined by hierarchical generalized linear model (HGLM) using a hospital-level random-intercept model. The mean medical cost in the last six months of life was $2,611±3,329 (U.S. dollars). In HGLM using a random-intercept model, we found that patients younger than 65 years had an additional cost of $819 over those aged ≥65 years. Patients who had a high Charlson Comorbidity Index Score (CCIS) had an additional $616 cost over those with a low CCIS. Those who survived post-diagnosis less than 6 months had an additional $659 in expenses over those who survived more than 24 months. Medical cost was $249 more for patients who had medium to high individual SES, and $319 more for those who were treated by non-oncologists. This study provides useful information for decision makers in understanding end-of-life expenditure in oral cancer. We found significantly increased end-of-life expenditure in patients if they were younger than 65 years or treated by non-oncologists, or had high CCIS, medium to high individual SES, and survival of less than 6 months after diagnosis.


Regulation of measles virus gene expression by P protein coiled-coil properties.
The polymerase of negative-stranded RNA viruses consists of the large protein (L) and the phosphoprotein (P), the latter serving both as a chaperon and a cofactor for L. We mapped within measles virus (MeV) P the regions responsible for binding and stabilizing L and showed that the coiled-coil multimerization domain (MD) of P is required for gene expression. MeV MD is kinked as a result of the presence of a stammer. Both restoration of the heptad regularity and displacement of the stammer strongly decrease or abrogate activity in a minigenome assay. By contrast, P activity is rather tolerant of substitutions within the stammer. Single substitutions at the "a" or "d" hydrophobic anchor positions with residues of variable hydrophobicity revealed that P functionality requires a narrow range of cohesiveness of its MD. Results collectively indicate that, beyond merely ensuring P oligomerization, the MD finely tunes viral gene expression through its cohesiveness.


Catecholamine enzymes in the degenerative neurological disease idiopathic orthostatic hypotension.
Discrete brain areas and sympathetic ganglia obtained at autopsy from patients with idiopathic orthostatic hypotension were assayed for tyrosine hydroxylase and dopamine beta-hydroxylase. Dopamine beta-hydroxylase activity was decreased 7.5-fold in sympathetic ganglia, while tyrosine hydroxylase activity was reduced more than 50-fold in the pontine nucleus locus coeruleus. These observations indicate that noradrenergic neurons of both brain and ganglion are affected in idiopathic orthostatic hypotension, but suggest that the central and peripheral biochemical deficits differ.


A low-voltage electrokinetic nanochannel drug delivery system.
Recent work has elucidated the potential of important new therapeutic paradigms, including metronomic delivery and chronotherapy, in which the precise timing and location of therapeutic administration has a significant impact on efficacy and toxicity. New drug delivery architectures are needed to not only release drug continuously at precise rates, but also synchronize their release with circadian cycles. We present an actively controlled nanofluidic membrane that exploits electrophoresis to control the magnitude, duration, and timing of drug release. The membrane, produced using high precision silicon fabrication techniques, has platinum electrodes integrated at the inlet and outlet that allow both amplification and reversal of analyte delivery with low applied voltage (at or below 2 VDC). Device operation was demonstrated with solutions of both fluorescein isothiocyanate conjugated bovine serum albumin and lysozyme using fluorescence spectroscopy, fluorescence microscopy, and a lysozyme specific bio-assay and has been characterized for long-term molecular release and release reversibility. Through a combination of theoretical and experimental analysis, the relative contributions of electrophoresis and electroosmosis have been investigated. The membrane's clinically relevant electrophoretic release rate at 2 VDC exceeds the passive release by nearly one order of magnitude, demonstrating the potential to realize the therapeutic paradigm goal.


Periostin differentially induces proliferation, contraction and apoptosis of primary Dupuytren's disease and adjacent palmar fascia cells.
Dupuytren's disease, (DD), is a fibroproliferative condition of the palmar fascia in the hand, typically resulting in permanent contracture of one or more fingers. This fibromatosis is similar to scarring and other fibroses in displaying excess collagen secretion and contractile myofibroblast differentiation. In this report we expand on previous data demonstrating that POSTN mRNA, which encodes the extra-cellular matrix protein periostin, is up-regulated in Dupuytren's disease cord tissue relative to phenotypically normal palmar fascia. We demonstrate that the protein product of POSTN, periostin, is abundant in Dupuytren's disease cord tissue while little or no periostin immunoreactivity is evident in patient-matched control tissues. The relevance of periostin up-regulation in DD was assessed in primary cultures of cells derived from diseased and phenotypically unaffected palmar fascia from the same patients. These cells were grown in type-1 collagen-enriched culture conditions with or without periostin addition to more closely replicate the in vivo environment. Periostin was found to differentially regulate the apoptosis, proliferation, alpha smooth muscle actin expression and stressed Fibroblast Populated Collagen Lattice contraction of these cell types. We hypothesize that periostin, secreted by disease cord myofibroblasts into the extra-cellular matrix, promotes the transition of resident fibroblasts in the palmar fascia toward a myofibroblast phenotype, thereby promoting disease progression.


High computed tomographic correlations between carotid canal dehiscence and high jugular bulb in the middle ear.
Both carotid canal dehiscence (CCD) and high jugular bulb (HJB) are thought to increase the potential for disastrous consequences during middle ear surgery. Clinical co-presentation of these two great vessel variants has not yet been described. This study aims to determine the relationship between CCD and HJB based on a computed tomographic (CT) temporal bone evaluation. High-resolution CT scans of 408 temporal bones obtained from 208 adults were recruited. Carotid canal integrity, jugular bulb position, petrous apex pneumatization and the minimal thickness of the carotid canal wall (TCW) facing the tympanic cavity were examined and measured for the incidence of CCD and/or HJB. Other variables including gender, age, laterality and the presence of otitis media or mastoiditis were also collected for analysis. CCD was found in 28 ears (6.9%); 19 of these were found to also have HJB (67.9%). The presence of CCD was significantly correlated with HJB presentation. The minimal TCW in HJB ears was significantly thinner than that of normally positioned jugular bulbs. Moreover, after controlling for other candidate variables, the independent factors of age (younger or older than 50 years) and jugular bulb position (high vs. normal) were found to predict the presence of CCD. In conclusion, HJB tends to coexist with a thinner carotid canal wall. This finding emphasizes the need to be watchful for the coexistence of these two great vessel anomalies when surgeons encounter an aged patient presenting either CCD or HJB during middle ear surgery.


Moxidectin has a lower neurotoxic potential but comparable brain penetration in P-glycoprotein-deficient CF-1 mice compared to ivermectin.
The anti-parasitic drugs ivermectin (IVM) and moxidectin (MOX) normally show limited brain penetration in vertebrates because of effective drug efflux at the blood-brain barrier by P-glycoprotein, encoded by the multi-drug resistance (MDR1) gene. However, dogs with homozygous nt230(del4) mutation in the MDR1 gene do not express a functionally active P-glycoprotein and show increased brain penetration of these drugs, resulting in neurological toxicity to different degrees. Thus, whereas IVM provokes neurological toxicity at 0.1 mg/kg, MOX is tolerated at this dosage. To investigate whether this difference is attributable to lower brain penetration of MOX in the absence of P-glycoprotein or to their neurotoxic potential, we applied IVM and MOX to P-glycoprotein-deficient CF-1 mice and comparatively analysed the absolute drug concentrations in the brain. Furthermore, we quantified drug-induced neurotoxicity by measuring the walking performance of the mice on a rotarod setup. We found that at a dosage of 0.2 mg/kg, representing 0.23 μmol/kg IVM and 0.31 μmol/kg MOX, the absolute drug concentrations in the brain were comparable with 100.8 pmol/g and 140.2 pmol/g, respectively. However, MOX induced the same degree of neurotoxicosis at the higher dosage of 1.09 μmol/kg (0.7 mg/kg) compared with IVM at 0.40 μmol/kg (0.35 mg/kg), demonstrating the 2.7-fold lower neurotoxic potential of MOX compared to IVM. This could be explained by a lower binding affinity or lower intrinsic activity of MOX at the relevant central nervous system receptors compared with IVM.


'As a man I felt small': a qualitative study of Ugandan men's experiences of living with a wife suffering from obstetric fistula.
The effects of obstetric fistula surpass the individual woman and affect husbands, relatives, peers and the community at large. Few studies have documented the experiences of men who live with wives suffering from fistula. In this study, our objective was to understand how fistula affects these men's lives. We conducted 16 in-depth interviews with men in central and western Uganda. We used thematic narrative analysis and discuss our findings based on Connell's theory of hegemonic masculinity. Findings show that the men's experiences conflicted with Ugandan norms of hegemonic masculinity. However, men had to find other ways of explaining their identity, such as portraying themselves as small men but still be responsible, caring husbands and fathers. The few individuals who married a second wife remained married to the wife with the fistula. These men viewed marriage as a lifetime promise before God and a responsibility that should not end because of a fistula. Poverty, love, care for children and social norms in a patriarchal society compelled the men to persevere in their relationship amidst many challenges.


Investigating breast cancer cell behavior using tissue engineering scaffolds.
Despite early detection through the use of mammograms and aggressive intervention, breast cancer (BC) remains a clinical dilemma. BC can resurge after >10 years of remission. Studies indicate that BC cells (BCCs) with self-renewal and chemoresistance could be involved in dormancy. The majority of studies use in vitro, two-dimensional (2-D) monolayer cultures, which do not recapitulate the in vivo microenvironment. Thus, to determine the effect of three-dimensional (3-D) microenvironment on BCCs, this study fabricated tissue engineering scaffolds made of poly (ε-caprolactone) (PCL) having aligned or random fibers. Random and aligned fibers mimic, respectively, the random and highly organized collagen fibers found in the tumor extracellular matrix. Chemoresistant BCCs were obtained by treating with carboplatin. Western blot analysis of carboplatin resistant (treated) MDA-MB-231 (highly invasive, basal-like) and T47D (low-invasive, luminal) BCCs showed an increase in Bcl-2, Oct-4 and Sox-2, suggesting protection from apoptosis and increase in stem-like markers. Further studies with MDA-MB-231 BCCs seeded on the scaffolds showed little to no change in cell number over time for non-treated BCCs whereas on tissue culture polystyrene (TCP), non-treated BCCs displayed a significant increase in cell number at days 4 and 7 as compared to day 1 (p<0.05). Treated BCCs did not proliferate on TCP and the fibrous scaffolds. Little to no cyclin D1 was expressed for non-treated BCCs on TCP. On fibrous scaffolds, non-treated BCCs stained for cyclin D1 during the 7-day culture period. Treated BCCs expressed cyclin D1 on TCP and fibrous scaffolds during the 7-day culture period. Proliferation, viability and cell cycle analysis indicated that this 3-D culture prompted the aggressive BCCs to adopt a dormant phenotype, while the treated BCCs retained their phenotype. The findings indicate that random and aligned fibrous PCL scaffolds may provide a useful system to study how the 3-D microenvironment affects the behavior of BCCs.


Retell as an Indicator of Reading Comprehension.
The purpose of this narrative synthesis is to determine the reliability and validity of retell protocols for assessing reading comprehension of students in grades K-12. Fifty-four studies were systematically coded for data related to the administration protocol, scoring procedures, and technical adequacy of the retell component. Retell was moderately correlated with standardized measures of reading comprehension and, with older students, had a lower correlation with decoding and fluency. Literal information was retold more frequently than inferential, and students with learning disabilities or reading difficulties needed more supports to demonstrate adequate recall. Great variability was shown in the prompting procedures, but scoring methods were more consistent across studies. The influences of genre, background knowledge, and organizational features were often specific to particular content, texts, or students. Overall, retell has not yet demonstrated adequacy as a progress monitoring instrument.


Molecular dynamics simulation study of the binding of purine bases to the aptamer domain of the guanine sensing riboswitch.
Riboswitches are a novel class of genetic control elements that function through the direct interaction of small metabolite molecules with structured RNA elements. The ligand is bound with high specificity and affinity to its RNA target and induces conformational changes of the RNA's secondary and tertiary structure upon binding. To elucidate the molecular basis of the remarkable ligand selectivity and affinity of one of these riboswitches, extensive all-atom molecular dynamics simulations in explicit solvent (approximately 1 micros total simulation length) of the aptamer domain of the guanine sensing riboswitch are performed. The conformational dynamics is studied when the system is bound to its cognate ligand guanine as well as bound to the non-cognate ligand adenine and in its free form. The simulations indicate that residue U51 in the aptamer domain functions as a general docking platform for purine bases, whereas the interactions between C74 and the ligand are crucial for ligand selectivity. These findings either suggest a two-step ligand recognition process, including a general purine binding step and a subsequent selection of the cognate ligand, or hint at different initial interactions of cognate and noncognate ligands with residues of the ligand binding pocket. To explore possible pathways of complex dissociation, various nonequilibrium simulations are performed which account for the first steps of ligand unbinding. The results delineate the minimal set of conformational changes needed for ligand release, suggest two possible pathways for the dissociation reaction, and underline the importance of long-range tertiary contacts for locking the ligand in the complex.


Rapid and reversible formation of spine head filopodia in response to muscarinic receptor activation in CA1 pyramidal cells.
A key feature at excitatory synapses is the remodelling of dendritic spines, which in conjunction with receptor trafficking modifies the efficacy of neurotransmission. Here we investigated whether activation of cholinergic receptors, which can modulate synaptic plasticity, also mediates changes in dendritic spine structure. Using confocal time-lapse microscopy in mouse slice cultures we found that brief activation of muscarinic receptors induced the emergence of fine filopodia from spine heads in all CA1 pyramidal cells examined. This response was widespread occurring in 48% of imaged spines, appeared within minutes, was reversible, and was blocked by atropine. Electron microscopic analyses showed that the spine head filopodia (SHFs) extend along the presynaptic bouton. In addition, the decay time of miniature EPSCs was longer after application of the muscarinic acetylcholine receptor agonist methacholine (MCh). Both morphological and electrophysiological changes were reduced by preventing microtubule polymerization with nocodazole. This extension of SHFs during cholinergic receptor activation represents a novel structural form of subspine plasticity that may regulate synaptic properties by fine-tuning interactions between presynaptic boutons and dendritic spines.


Neurostimulation as a Method of Treatment and a Preventive Measure in Canine Drug-Resistant Epilepsy: Current State and Future Prospects.
Modulation of neuronal activity for seizure control using various methods of neurostimulation is a rapidly developing field in epileptology, especially in treatment of refractory epilepsy. Promising results in human clinical practice, such as diminished seizure burden, reduced incidence of sudden unexplained death in epilepsy, and improved quality of life has brought neurostimulation into the focus of veterinary medicine as a therapeutic option. This article provides a comprehensive review of available neurostimulation methods for seizure management in drug-resistant epilepsy in canine patients. Recent progress in non-invasive modalities, such as repetitive transcranial magnetic stimulation and transcutaneous vagus nerve stimulation is highlighted. We further discuss potential future advances and their plausible application as means for preventing epileptogenesis in dogs.


The p38 MAP kinase pathway as a therapeutic target in inflammatory disease.
The p38 MAPK signalling pathway plays an important role in inflammation and other physiological processes. Specific inhibitors of p38 alpha and beta MAPK block production of the major inflammatory cytokines (i.e. tumour necrosis factor-alpha and interleukin-1) and other proteins (e.g. cyclooxygenase-2), and are anti-inflammatory in animal models of disease. A major function of the pathway is post-transcriptional control of inflammatory gene expression. Many of the mRNAs are unstable (or untranslatable) because of AU-rich elements in the 3'untranslated region. Signalling in the p38 pathway counteracts these and stabilizes the mRNAs by preventing their otherwise rapid de-adenylation.


Effects of six chemical deicers on larval wood frogs (Rana sylvatica).
Widespread and intensive application of road deicers, primarily road salt (NaCl), in North America threatens water quality and the health of freshwater ecosystems. Intensive use of NaCl can be harmful to sensitive members of freshwater ecosystems such as amphibians. Detection of negative effects of NaCl application has prompted the search for alternative chemical deicers with lower environmental impacts. We conducted a series of 96-h acute toxicity tests to determine the negative sensitivity of larval wood frogs (Rana [Lithobates] sylvatica) to six deicing chemicals: urea (CH(4) N(2) O), sodium chloride (NaCl), magnesium chloride (MgCl(2) ), potassium acetate (CH(3) COOK), calcium chloride (CaCl(2) ), and calcium magnesium acetate (C(8) H(12) CaMgO(8) ). Acetates are sometimes touted as environmentally friendly alternatives to NaCl but have not been examined in enough detail to warrant this designation. When exposed to a range of environmentally realistic concentrations of these chemicals, larvae were least sensitive (i.e., had the lowest mortality rate) to CH(4) N(2) O, NaCl, and MgCl(2) and most sensitive to acetates (C(8) H(12) CaMgO(8) , CH(3) COOK) and CaCl(2) . Our observed median lethal concentration estimates (LC50(96-h) ) for NaCl were over two times higher than values presented in previous studies, which suggests variability in tolerance among R. sylvatica populations. The deicers varied greatly in their toxicity, and further research is warranted to examine the differential effects of this suite of deicers on other species.


Human antilipid A monoclonal antibodies bind to human B cells and the i antigen on cord red blood cells.
We describe two independently derived human mAb, A6(H4C5) and 216, initially selected for their reactivity to the lipid A domain of bacterial LPS, which also react with the following Ag: the i Ag present on cord RBC, a ligand on human B lymphocytes, and to certain autoantigens, defining these mAb as polyreactive. Both mAb have specific affinity for a carbohydrate epitope consisting minimally of a disaccharide with an acyl substitution at the 2-carbon position. Structural examination of the diverse Ag recognized by the two antibodies reveals the presence of this carbohydrate structure required for antibody binding. A6(H4C5) and 216 are IgM in isotype, but differ in their L chain expression. Molecular analysis shows that both the mAb are encoded by a highly conserved VH4 gene, designated VH4-21. This gene encodes a number of autoantibodies, particularly cold agglutinins. Specific recognition of lipid A and of a carbohydrate epitope on B lymphocytes by the two human mAb suggests a dual function for the highly conserved VH4-21 gene in antibacterial response and in B cell development and regulation.


The impact of stressors on second generation Indian Residential School survivors.
From 1863 to 1996, many Aboriginal children in Canada were forced to attend Indian Residential Schools (IRSs), where many experienced neglect, abuse, and the trauma of separation from their families and culture. The present study examined the intergenerational impact of IRS exposure on depressive symptomatology in a convenience sample of 143 First Nations adults. IRS experiences had adverse intergenerational effects in that First Nations adults who had a parent attend IRS (n = 67) reported greater depressive symptoms compared to individuals whose parents did not attend (n = 76). Parental IRS attendance moderated the relations between stressor experiences (adverse childhood experiences, adult traumas, and perceived discrimination) and depressive symptoms, such that second generation Survivors exhibited greater symptomatology. Adverse childhood experiences partially mediated the relation between parental IRS attendance and both adult trauma and perceived discrimination. Moreover, both of these adulthood stressors partially mediated the relation between adverse childhood experiences and depressive symptoms. Finally, all three stressors demonstrated a unique mediating role in the relation between parental IRS attendance and depressive symptoms. Although alternative directional paths could not be ruled out, offspring of IRS Survivors appeared at increased risk for depression, likely owing to greater sensitivity to and experiences of childhood adversity, adult traumas, and perceived discrimination.


Fluorescence spectroscopy for identification of atherosclerotic tissue.
Vessel perforation and limited steerability of the laser light are the major limitations of laser angioplasty. To improve steerability fluorescence spectroscopy has been proposed for identification of atherosclerotic plaques. The aim was to investigate this. Fluorescence spectroscopy with three different excitation wavelengths (325 nm, 380 nm, 450 nm) was tested in an emission range of 400 nm to 600 nm. Intensity ratios at 480/420 nm were determined in different types of blood vessels. Necropsy material from 40 patients (punch biopsies of 4 mm diameter from the coronary and carotid artery as well as from the ascending and descending aorta) was studied spectroscopically. Histological alterations of the vessel wall were assessed by a semiquantitative score (0 to 10 points): (a) normal tissue, 0 to 2 points (mean = 0.25; n = 38); (b) mild atherosclerotic lesions, 3 to 5 points (mean = 3.35; n = 39); (c) severe atherosclerotic lesions, greater than or equal to 6 points (mean = 6.75; n = 43). Best spectroscopic results were obtained with an excitation wavelength of 325 nm. In samples with severe atherosclerotic lesions the fluorescence spectra showed a significant reduction of the emitted wavelength intensities when compared to normal tissue. There was a clear separation of the fluorescence spectra between normal and mild as well as between normal and severe atherosclerotic lesions; normal tissue showed an increased intensity in the range from 420 nm to 540 nm, whereas atherosclerotic lesions had no or only a small peak at 480 nm. There was a significant correlation between the semiquantitative score (n = 120) and the fluorescence ratio at 480/420 nm (excitation wavelength 325 nm) with a correlation coefficient of 0.87. The spectroscopic results showed no differences between the samples taken from different types of vessels. Fluorescence spectroscopy allows a reliable identification of normal and atherosclerotic lesions. The close correlation between the emitted light intensity ratio at 480/420 nm and the histological alterations of the vessel wall suggests a relationship between vessel wall fluorescence and the atherosclerotic alterations of the wall.


Development of a questionnaire to assess worker knowledge, attitudes and perceptions underlying dermal exposure.
Workers' behavior is identified as an important determinant of dermal exposure and is influenced by knowledge, attitudes, and risk perceptions. Because behavior may be a significant predictor of exposure, its assessment provides a means for examining exposure and designing strategies and incentives that encourage worker protective behavior. Currently, there are no psychosocial instruments examining worker knowledge, attitudes, and perceptions with respect to dermal hazards. Accordingly, a questionnaire was developed and tested to provide an instrument for measuring worker knowledge, attitudes, and perceptions. The questionnaire was developed on the basis of a literature review and expert consultation. Scales were constructed based on standard methods. Two worker focus groups were used to evaluate worker understanding and content validity of the KAP (knowledge, attitudes, perceptions) questionnaire. The resulting 115-item questionnaire that included scales for knowledge (N=13), attitudes (N=27), perceptions (N=15), behavior (N=8), behavioral intentions (N=15), barriers (N=13), and facilitators (N=5) was tested on 89 workers from 19 facilities. The concepts identified in the focus groups included worker perception of higher risk due to a poor fit and replacement frequency for personal protective equipment and cross-contamination by workers moving into work zones. Field testing of the questionnaire (N=89) yielded Cronbach's alpha reliability scores ranging from 0.87 for the self-efficacy personal protective equipment scale to 0.92 for the overall belief scale, indicating high internal reliability. Although further testing and refinement is needed, this survey instrument provides an initial and conceptually unique means for evaluating behavioral determinants of worker dermal exposure.


Predicting functionally informative mutations in Escherichia coli BamA using evolutionary covariance analysis.
The essential outer membrane β-barrel protein BamA forms a complex with four lipoprotein partners BamBCDE that assembles β-barrel proteins into the outer membrane of Escherichia coli. Detailed genetic studies have shown that BamA cycles through multiple conformations during substrate assembly, suggesting that a complex network of residues may be involved in coordinating conformational changes and lipoprotein partner function. While genetic analysis of BamA has been informative, it has also been slow in the absence of a straightforward selection for mutants. Here we take a bioinformatic approach to identify candidate residues for mutagenesis using direct coupling analysis. Starting with the BamA paralog FhaC, we show that direct coupling analysis works well for large β-barrel proteins, identifying pairs of residues in close proximity in tertiary structure with a true positive rate of 0.64 over the top 50 predictions. To reduce the effects of noise, we designed and incorporated a novel structured prior into the empirical correlation matrix, dramatically increasing the FhaC true positive rate from 0.64 to 0.88 over the top 50 predictions. Our direct coupling analysis of BamA implicates residues R661 and D740 in a functional interaction. We find that the substitutions R661G and D740G each confer OM permeability defects and destabilize the BamA β-barrel. We also identify synthetic phenotypes and cross-suppressors that suggest R661 and D740 function in a similar process and may interact directly. We expect that the direct coupling analysis approach to informed mutagenesis will be particularly useful in systems lacking adequate selections and for dynamic proteins with multiple conformations.


Dissociable Catecholaminergic Modulation of Visual Attention: Differential Effects of Catechol-O-Methyltransferase and Dopamine Beta-Hydroxylase Genes on Visual Attention.
Visual attention enables us to prioritise behaviourally relevant visual information while ignoring distraction. The neural networks supporting attention are modulated by two catecholamines, dopamine and noradrenaline. The current study investigated the effects of single nucleotide polymorphisms in two catecholaminergic genes - COMT (Val158Met) and DBH (444 G/A) - on individual differences in attention functions. Participants (n = 125) were recruited from the Oxford Biobank by genotype-based recall. They were tested on a continuous performance task (sustained attention), a Go/No-Go task (response inhibition), and a task assessing attentional selection in accordance with the Theory of Visual Attention (TVA). We found a significant effect of DBH genotype status on the capacity to maintain attention over time (sustained attention) as measured by the continuous performance task. Furthermore, we demonstrated a significant association between COMT genotype status and effective threshold of visual perception in attentional selection as estimated based on the TVA task performance. No other group differences in attention function were found with respect to the studied genotypes. Overall, our findings provide novel experimental evidence that: (i) dopaminergic and noradrenergic genotypes have dissociable effects on visual attention; (ii) either insufficient or excessive catecholaminergic activity may have equally detrimental effects on sustained attention.


ACELIFT: a minimally invasive alternative to a facelift.
Cervicofacial aging is often characterized by a combination of skin and subcutaneous tissue laxity, midfacial deflation, an accumulation of excess submental fat, an obtuse cervicomental angle, jowls, and rhytides of the face and neck. Traditional treatment, and the "gold standard" against which other treatments are compared, is a facelift. To demonstrate that a combination technique called ACELIFT - an acronym for the Augmentation of Collagen and Elastin using Lasers, Injectable neurotoxins, Fillers, and Topicals - in selected patients, is a viable, safe, and effective alternative to a facelift. Ten healthy women, ages 50 to 62 (mean age = 58), with cervical and facial stigmata of aging were enrolled in a prospective study conducted in the authors' private practice. Patients underwent a two-step procedure; the first step was laser lipolysis of the submental and anterior cervical areas with a pulsed 1440 nm Nd:YAG laser with a side-firing fiber (PrecisionTx, Cynosure, Westford, MA). Three months later, the patients were treated in a single session that combined injectable neurotoxin, fillers, and fractional (Fx) CO2 laser resurfacing delivered in a novel "hammock" distribution. After two weeks, following complete re-epithelialization, the patients were started on a topical regimen that included daily use of sunscreen and antioxidants and nightly use of retinoids and peptides. This regimen was continued for a period of six months when all patients returned for final evaluation. Nine months following the initiation of treatment, all patients were evaluated by the following: Global Aesthetic Improvement Scale, cervicomental angle scale, physician, and subject evaluation. Clinical improvement was evident, and often marked, for all subjects. Both physician and subject satisfaction scores were high, indicating overall satisfaction with the procedure and the outcomes. Side-effects were mild and transient; there were no incidents of adverse scarring, thermal injuries, permanent nerve injury, or dyschromia, hematomas, seromas, or infection. Subjects were likely to recommend the procedure to a friend. In properly selected patients, the ACELIFT proved to be a safe and effective, minimally invasive alternative to a facelift. There was little downtime and high patient satisfaction.


Assessment of antifouling effectiveness of two natural product antifoulants by attachment study with freshwater bacteria.
GOAL, SCOPE, BACKGROUND: The traditional solution for keeping unwanted organisms from attaching to submerged surfaces is to apply anti-fouling coatings. The most common antifoulant was tributyltin (TBT). TBT systems were highly effective but were also toxic to non-target organisms. The use of the TBT based coatings will be completely banned by January 1, 2008. Therefore, there is an urgent need to seek out suitable non-toxic alternatives. The aim of this work was to evaluate the effectiveness of capsaicin and zosteric acid as natural product antifoulants (NPAs) in deterring bacterial attachment. Two fresh water bacteria systems Pseudomonas putida (Pp) and bacteria isolated from Lake Erie (LE) were used to assess the attachment when the NPAs dispersed in the water. Effectiveness was ascertained based on the decrease in microbial attachment, limited toxicity, and minimum alteration of the coatings properties. A significant inhibition of bacteria attachment was achieved when aqueous capsaicin concentration was increased from 0 to 40 mg/L. For instance, after 14 days the LE system depicted 93.5% and 98.5% less biofilm coverage for 20 mg/L and 40 mg/L capsaicin, respectively when pared to systems without NPA. Biofilm coverage was reduced by 92.5% and 98.2%, respectively with 50 mg/L and 500 mg/L zosteric acid. Both capsaicin and zosteric acid was effective at preventing bacteria attachment. As the NPA aqueous concentration increased, biofilm formation decreased. Evaluating changes in aqueous pH, conductivity, dissolved oxygen, aqueous microbial population and biofilm formation suggested that the primary antifoulant mechanism of these two NPAs was to block the bacteria's active sites versus posing a lethal level. From the attachment study, zosteric acid appeared to be more effective in preventing bacterial attachment when the NPAs were dispersed in the aqueous environment. For practical applications, the antifoulant needs to be incorporated into a coating and have a slow release rate. Thus the ability to successfully incorporate zosteric acid into a coating, without deterring bacterial attachment, needs to be investigated.