Collybolide is a novel biased agonist of κ-opioid receptors with potent antipruritic activity.
Among the opioid receptors, the κ-opioid receptor (κOR) has been gaining considerable attention as a potential therapeutic target for the treatment of complex CNS disorders including depression, visceral pain, and cocaine addiction. With an interest in discovering novel ligands targeting κOR, we searched natural products for unusual scaffolds and identified collybolide (Colly), a nonnitrogenous sesquiterpene from the mushroom Collybia maculata. This compound has a furyl-δ-lactone core similar to that of Salvinorin A (Sal A), another natural product from the plant Salvia divinorum Characterization of the molecular pharmacological properties reveals that Colly, like Sal A, is a highly potent and selective κOR agonist. However, the two compounds differ in certain signaling and behavioral properties. Colly exhibits 10- to 50-fold higher potency in activating the mitogen-activated protein kinase pathway compared with Sal A. Taken with the fact that the two compounds are equipotent for inhibiting adenylyl cyclase activity, these results suggest that Colly behaves as a biased agonist of κOR. Behavioral studies also support the biased agonistic activity of Colly in that it exhibits ∼10-fold higher potency in blocking non-histamine-mediated itch compared with Sal A, and this difference is not seen in pain attenuation by these two compounds. These results represent a rare example of functional selectivity by two natural products that act on the same receptor. The biased agonistic activity, along with an easily modifiable structure compared with Sal A, makes Colly an ideal candidate for the development of novel therapeutics targeting κOR with reduced side effects.


Comparison of hepatic venous pressure gradient and two models of end-stage liver disease for predicting the survival in patients with decompensated liver cirrhosis.
We evaluated the efficacy of initial and follow-up hepatic venous pressure gradient (HVPG), models of end-stage liver disease (MELD), and MELD-Na for predicting the survival of patients with decompensated liver cirrhosis (LC). MELD with/without Na score and HVPG have been important predictors of mortality in patients with LC. Between January 2006 and 2011, a total of 57 patients with decompensated LC, all of whom underwent >2 HVPG measurements for the confirmation of propranolol dosing, were enrolled. MELD and MELD-Na scores were calculated on the day of HVPG measurement. The prognostic accuracy of the initial and follow-up HVPG, MELD, and MELD-Na were analyzed, and independent factors for mortality were evaluated. Ten patients (17.5%) died from LC. Initial HVPG (0.883), initial MELD-Na (0.877), follow-up HVPG (0.829), and follow-up MELD-Na (0.802) showed good area under the receiver operating characteristic curve scores in predicting 1-year mortality. In predicting 2-year mortality, only follow-up HVPG (0.821, cut-off value 18 mm Hg) showed good score. Overall area under the receiver operating characteristic curves (initial and follow-up) were 0.843 and 0.864 in HVPG, 0.721 and 0.674 in MELD, and 0.762 and 0.715 in MELD-Na, respectively. In the Cox regression analysis, only follow-up HVPG (P=0.02; odds ratio, 1.11) was associated with mortality. The efficacy of HVPG for predicting mortality is excellent compared with that of MELD or MELD-Na. Therefore, aside from the confirmation of adequate propranolol dosing, HVPG may be needed for predicting the survival of patients with decompensated LC.


Editor's Choice-The role of the emergency department in the management of acute heart failure: An international perspective on education and research.
Emergency departments are a major entry point for the initial management of acute heart failure (AHF) patients throughout the world. The initial diagnosis, management and disposition - the decision to admit or discharge - of AHF patients in the emergency department has significant downstream implications. Misdiagnosis, under or overtreatment, or inappropriate admission may place patients at increased risk for adverse events, and add costs to the healthcare system. Despite the critical importance of initial management, data are sparse regarding the impact of early AHF treatment delivered in the emergency department compared to inpatient or chronic heart failure management. Unfortunately, outcomes remain poor, with nearly a third of patients dying or re-hospitalised within 3 months post-discharge. In the absence of robust research evidence, consensus is an important source of guidance for AHF care. Thus, we convened an international group of practising emergency physicians, cardiologists and advanced practice nurses with the following goals to improve outcomes for AHF patients who present to the emergency department or other acute care setting through: (a) a better understanding of the pathophysiology, presentation and management of the initial phase of AHF care; (b) improving initial management by addressing knowledge gaps between best practices and current practice through education and research; and


Proof-testing and probabilistic lifetime estimation of glass fibers for sensor applications.
The mechanical reliability of sensing glass fiber is one of the important problems in the practical use of fiber-optic sensors. To ensure long-term reliability on a mass-production level, a method of proof-testing is applied to a sensing glass fiber that will be subjected to mechanical deformation in its service situation. We propose to employ a higher strain level (screening level) in the proof-testing with a fiber-recoating technique that can suppress excessive damage during the testing. We consider a standard lifetime of 15 years of automotive applications and ensure a practical level of failure probability by a model calculation by using the strength data of a prototype fiber with the method of fracture-mechanics theory.


A randomized trial comparing the efficacy and safety of intravitreal triamcinolone with standard care to treat vision loss associated with macular Edema secondary to branch retinal vein occlusion: the Standard Care vs Corticosteroid for Retinal Vein Occlusion (SCORE) study report 6.
To compare the efficacy and safety of 1-mg and 4-mg doses of preservative-free intravitreal triamcinolone with standard care (grid photocoagulation in eyes without dense macular hemorrhage and deferral of photocoagulation until hemorrhage clears in eyes with dense macular hemorrhage) for eyes with vision loss associated with macular edema secondary to branch retinal vein occlusion (BRVO). Multicenter, randomized clinical trial of 411 participants. Main Outcome Measure Gain in visual acuity letter score of 15 or more from baseline to month 12. Twenty-nine percent, 26%, and 27% of participants achieved the primary outcome in the standard care, 1-mg, and 4-mg groups, respectively. None of the pairwise comparisons between the 3 groups was statistically significant at month 12. The rates of elevated intraocular pressure and cataract were similar for the standard care and 1-mg groups, but higher in the 4-mg group. There was no difference identified in visual acuity at 12 months for the standard care group compared with the triamcinolone groups; however, rates of adverse events (particularly elevated intraocular pressure and cataract) were highest in the 4-mg group. Application to Clinical Practice Grid photocoagulation as applied in the SCORE Study remains the standard care for patients with vision loss associated with macular edema secondary to BRVO who have characteristics similar to participants in the SCORE-BRVO trial. Grid photocoagulation should remain the benchmark against which other treatments are compared in clinical trials for eyes with vision loss associated with macular edema secondary to BRVO. Trial Registration clinicaltrials.gov Identifier: NCT00105027.


Attenuation of rabies virulence: takeover by the cytoplasmic domain of its envelope protein.
The capacity of a rabies virus to promote neuronal survival (a signature of virulence) or death (a marker of attenuation) depends on the cellular partners recruited by the PDZ-binding site (PDZ-BS) of its envelope glycoprotein (G). Neuronal survival requires the selective association of the PDZ-BS of G with the PDZ domains of two closely related serine-threonine kinases, MAST1 and MAST2. Here, we found that a single amino acid change in the PDZ-BS triggered the apoptotic death of infected neurons and enabled G to interact with additional PDZ partners, in particular the tyrosine phosphatase PTPN4. Knockdown of PTPN4 abrogated virus-mediated apoptosis. Thus, we propose that attenuation of rabies virus requires expansion of the set of host PDZ proteins with which G interacts, which interferes with the finely tuned homeostasis required for survival of the infected neuron.


Cerebral hemodynamic failure presenting as limb-shaking transient ischemic attacks.
Limb-shaking transient ischemic attacks (TIA) may occur in patients with insufficient brain perfusion due to an underlying occlusive disease. We present the case of a 64-year-old patient who suffered from repetitive TIA presenting with shaking movements of the right-sided extremities and accompanying speech arrest. Symptoms are documented in the online supplementary video (www.karger.com/doi/10.1159/000327683). These episodes were frequently triggered in orthostatic situations. The diagnosis of limb-shaking TIA was established. The diagnostic workup revealed pseudo-occlusion of the left internal carotid artery, a poor intracranial collateral status and, as a consequence, an exhausted vasomotor reserve capacity. At ultrasound examination, symptoms were provoked by a change of the patient's position from supine to sitting. During evolvement of symptoms, a dramatic decrease of flow velocities in the left middle cerebral artery was observed. This case thus documents the magnitude and dynamics of perfusion failure in a rare manifestation of cerebral ischemic disease.


Why copy others? Insights from the social learning strategies tournament.
Social learning (learning through observation or interaction with other individuals) is widespread in nature and is central to the remarkable success of humanity, yet it remains unclear why copying is profitable and how to copy most effectively. To address these questions, we organized a computer tournament in which entrants submitted strategies specifying how to use social learning and its asocial alternative (for example, trial-and-error learning) to acquire adaptive behavior in a complex environment. Most current theory predicts the emergence of mixed strategies that rely on some combination of the two types of learning. In the tournament, however, strategies that relied heavily on social learning were found to be remarkably successful, even when asocial information was no more costly than social information. Social learning proved advantageous because individuals frequently demonstrated the highest-payoff behavior in their repertoire, inadvertently filtering information for copiers. The winning strategy (discountmachine) relied nearly exclusively on social learning and weighted information according to the time since acquisition.


Dried distillers grains plus solubles with corn silage or alfalfa hay as the primary forage source in dairy cow diets.
Nine multiparous (250 +/- 6 d in milk) and 3 primiparous (204 +/- 6 d in milk) Holstein cows were utilized in a 3 x 3 Latin square design to evaluate the lactation performance of cows fed a diet containing dried distillers grains plus solubles (DDGS) with either corn silage or alfalfa hay as forage. Cows were fed total mixed diets containing corn silage (CS), 50% corn silage and 50% alfalfa hay (CSAH), or alfalfa hay (AH) as the forage source. All diets had a 50:50 forage-to-concentrate ratio, contained 15% DDGS, and were formulated to be equal in metabolizable protein. Dry matter intake increased when cows were fed CSAH (24.9 kg/d) compared with CS (21.9 kg/d) and AH (20.9 kg/d). Yields of milk (26.5, 28.4, 29.0 kg/d for CS, CSAH, and AH, respectively) increased linearly as proportions of alfalfa fed increased but 4% fat-corrected milk and energy-corrected milk were not affected by treatment. Feed efficiency (1.28, 1.23, and 1.45 kg of energy-corrected milk/kg of intake) improved when AH was fed compared with CS or CSAH. Milk fat concentration (3.67, 3.55, and 3.49%) decreased linearly when alfalfa replaced corn silage, but was observed only in primiparous cows, not multiparous cows. Milk protein concentration (3.32, 3.29, and 3.29%) was not affected by diet although yield (0.90, 0.96, and 0.98 kg/d) tended to increase linearly when alfalfa was added to the diet. This may have been due to an increase in essential amino acid (AA) availability and uptake by the mammary gland or to greater crude protein intake in cows fed AH. In addition, replacing corn silage with alfalfa increased the uptake of Lys by the mammary gland. Methionine was the first-limiting AA based on the transfer efficiency of AA in arterial plasma to milk protein. However, Lys was the first-limiting AA in CS and CSAH and Met was first limiting in AH for mammary gland extraction efficiency of AA from plasma. In conclusion, replacing corn silage with alfalfa hay in diets containing 15% DDGS increased milk yield and tended to increase milk protein yield linearly in cows during late lactation. Feeding alfalfa hay as the sole forage source improved feed efficiency compared with diets containing corn silage.


Development and Evaluation of Melt-in-Mouth Tablets of Metoclopramide Hydrochloride Using Novel Co-processed Superdisintegrants.
In the present investigation, a novel multifunctional co-processed superdisintegrants consisting of crospovidone and Kyron T-314 were fabricated by solvent evaporation method to develop melt-in-mouth tablets of metoclopramide hydrochloride with a view to enhance patient compliance by direct compression method. The simple physical blends and co-processed mixture of superdisintegrants were characterized for angle of repose, bulk density, tapped density, Carr's index, Hausner's ratio and compatibility studies by FTIR spectroscopy. Melt-in-mouth tablets of metoclopramide hydrochloride were prepared using the physical blends and co-processed mixture of superdisinterants and were evaluated for hardness, friability, in vitro disintegration time, in vitro dispersion time, wetting time, water absorption ratio, drug content, in vitro drug release and accelerated stability study at 40±2° temperature and 75±5% relative humidity. Among the tablets evaluated, formulation F-X prepared by adding co-processed superdisintegrants in ratio of 1:1 showed minimum in vitro dispersion time of 9.71±0.021 s, in vitro disintegration time of 5.70±0.117 s and higher amount of drug release of 99.695±0.29% at the end of 1 min. Formulation F-X was emerged as the overall best formulation based on drug release characteristics in pH 6.8 phosphate buffer compared with the tablets obtained from conventional method of manufacture as well as with marketed preparation. Analysis of drug release data indicated that formulation F-X followed first order kinetics. This study revealed that the co-processed mixture of superdisintegrants have excellent flow properties, high compressibility, render low disintegration time to tablets and have better binding properties as compared to physical blends of superdisintegrants. These materials can be a good substitute for inert superdisintegrants, which are normally used in tablet manufacturing.


Darwinian aesthetics: sexual selection and the biology of beauty.
Current theoretical and empirical findings suggest that mate preferences are mainly cued on visual, vocal and chemical cues that reveal health including developmental health. Beautiful and irresistible features have evolved numerous times in plants and animals due to sexual selection, and such preferences and beauty standards provide evidence for the claim that human beauty and obsession with bodily beauty are mirrored in analogous traits and tendencies throughout the plant and animal kingdoms. Human beauty standards reflect our evolutionary distant and recent past and emphasize the role of health assessment in mate choice as reflected by analyses of the attractiveness of visual characters of the face and the body, but also of vocal and olfactory signals. Although beauty standards may vary between cultures and between times, we show in this review that the underlying selection pressures, which shaped the standards, are the same. Moreover we show that it is not the content of the standards that show evidence of convergence--it is the rules or how we construct beauty ideals that have universalities across cultures. These findings have implications for medical, social and biological sciences.


Is early removal of prophylactic ureteric stents beneficial in live donor renal transplantation?
Prophylactic ureteric stenting has been shown to reduce ureteric leaks and collecting system obstruction following renal transplantation and is in widespread use. However, the optimal time for removal of ureteric stents after renal transplantation remains unclear. Aim of this study was to compare the result of early versus late removal of ureteric stents after kidney transplantation of the laparoscopically retrieved live related donor grafts. Eligible patients were live donor kidney transplant recipients with normal urinary tracts. All recipients underwent extravesical Lich-Gregoire ureteroneocystostomy over 4F/160 cm polyurethane double J stents by a uniform technique. They were randomized on seventh postoperative day for early removal of stents on postoperative day 7 (Group I), or for late removal on postoperative day 28 (Group II). The incidence of urinary tract infections, asymptomatic bacteriuria, and urological complications were compared. Between 2007 and 2009, 130 kidney transplants were performed at one centre of which 100 were enrolled for the study, and 50 each were randomized into the two groups. Donor and recipient age, sex, native renal disease, immunosupression, number of rejection episodes, and antirejection therapy were similar in the two groups. The occurrence of symptomatic urinary tract infection during the follow-up period of 6 months was significantly less in the early stent removal group [5 out of 50 (10%) in Group I, vs 50 out of 15 (30%) in Group II, P=0.02]. Asymptomatic bacteriuria was documented in 2 out of 50 (4%) in Group I and 4 out of 50 (8%) in Group II (P=0.3). There was no statistically significant difference in the rate of ureteric leak, ureteric obstruction, or hematuria in the two groups (P=1.0). We conclude that, in kidney transplant recipients of laparoscopically retrieved live donor grafts, early stent removal at the end of first week reduces the incidence of urinary tract infection without increasing the rate of urine leak or ureteric obstruction.


A BEME systematic review of UK undergraduate medical education in the general practice setting: BEME Guide No. 32.
General practice is increasingly used as a learning environment in undergraduate medical education in the UK. The aim of this project was to identify, summarise and synthesise research about undergraduate medical education in general practice in the UK. We systematically identified studies of undergraduate medical education within a general practice setting in the UK from 1990 onwards. All papers were summarised in a descriptive report and categorised into two in-depth syntheses: a quantitative and a qualitative in-depth review. 169 papers were identified, representing research from 26 UK medical schools. The in-depth review of quantitative papers (n = 7) showed that medical students learned clinical skills as well or better in general practice settings. Students receive more teaching, and clerk and examine more patients in the general practice setting than in hospital. Patient satisfaction and enablement are similar whether a student is present or not in a consultation, however, patients experience lower relational empathy. Two main thematic groups emerged from the qualitative in-depth review (n = 10): the interpersonal interactions within the teaching consultation and the socio-cultural spaces of learning which shape these interactions. The GP has a role as a broker of the interactions between patients and students. General practice is a socio-cultural and developmental learning space for students, who need to negotiate the competing cultures between hospital and general practice. Lastly, patients are transient members of the learning community, and their role requires careful facilitation. General practice is as good, if not better, than hospital delivery of teaching of clinical skills. Our meta-ethnography has produced rich understandings of the complex relationships shaping possibilities for student and patient active participation in learning.


The Use of Indwelling Tunneled Pleural Catheters for Recurrent Pleural Effusions in Patients With Hematologic Malignancies: A Multicenter Study.
Malignant pleural effusion is a common complication of advanced malignancies. Indwelling tunneled pleural catheter (IPC) placement provides effective palliation but can be associated with complications, including infection. In particular, hematologic malignancy and the associated immunosuppressive treatment regimens may increase infectious complications. This study aimed to review outcomes in patients with hematologic malignancy undergoing IPC placement. A retrospective multicenter study of IPCs placed in patients with hematologic malignancy from January 2009 to December 2013 was performed. Inclusion criteria were recurrent, symptomatic pleural effusion and an underlying diagnosis of hematologic malignancy. Records were reviewed for patient demographics, operative reports, and pathology, cytology, and microbiology reports. Ninety-one patients (mean ± SD age, 65.4 ± 15.4 years) were identified from eight institutions. The mean × SD in situ dwell time of all catheters was 89.9 ± 127.1 days (total, 8,160 catheter-days). Seven infectious complications were identified, all of the pleural space. All patients were admitted to the hospital for treatment, with four requiring additional pleural procedures. Two patients died of septic shock related to pleural infection. We present, to our knowledge, the largest study examining clinical outcomes related to IPC placement in patients with hematologic malignancy. An overall 7.7% infection risk and 2.2% mortality were identified, similar to previously reported studies, despite the significant immunosuppression and pancytopenia often present in this population. IPC placement appears to remain a reasonable clinical option for patients with recurrent pleural effusions related to hematologic malignancy.


Boundaries to the articulation of possible selves through social networking sites: the case of Facebook profilers' social connectedness.
This study aims to contribute to an emerging literature that seeks to understand how identity markers on social networking sites (SNSs) shape interpersonal impressions, and particularly the boundaries that SNSs present for articulating unconstrained "hoped-for possible selves." An experiment employing mock-up Facebook profiles was conducted, showing that appearing with friends on a Facebook profile picture as well as increasingly higher number of Facebook friends strengthened perceptions of a profiler's hoped-for level of social connectedness. Excessive numbers of friends, however, weakened perceptions of a profiler's real-level social connectedness, particularly among participants with smaller social networks on Facebook themselves. The discussion focuses on when people come to find that reasonable boundaries of self-generated information on an SNS have been exceeded.


Optimal load maximizes the mean mechanical power output during upper extremity exercise in highly trained soccer players.
The purpose of this study was to determine the optimal load for the maximal power output during the acceleration phase of a power movement in bench press (BP) exercises of highly trained soccer players at the beginning of a competition period. Fifteen professional male soccer players with an average age of 26.1 ± 3.9 years, an average height of 183.3 ± 6.7 cm, an average body mass of 78.8 ± 7.2 kg, and an average 1 repetition maximum (1RM) of 83.3 ± 11.2 kg were employed as subjects in this study. Maximal mean power output during a BP at 0, 10, 30, 50, 70, and 90% of their 1RM was measured to determine whether an optimal load exists that allows for the attainment of maximal power output. Three-dimensional upper extremity kinematic data were collected. Two force plates embedded in the floor and positioned below the bench were used to measure contact forces between the bench and ground during the lift. A repeated-measures analysis of variance was performed to determine power output differences at different percentages of the 1RM. The results of this study indicated that loads of 50% of the 1RM resulted in greater mean power output during the complete positive power movement. Loads at 30 and 50% of the 1RM resulted in greater mean power output computed from the acceleration phase of the lift than did all loads and were not statistically different from each other. However, individual soccer players did not reach the maximum power output with the same relative load. In conclusion, when soccer players develop muscular power toward the end of when the most important competitions are scheduled, dynamic effort strength training with the range of load from 30 to 50% of 1RM BP should be used. During the competition period, a load of 50% of a 1RM should be used in a BP to maintain muscular power over a wide load range.


Intestinal organoids model human responses to infection by commensal and Shiga toxin producing Escherichia coli.
Infection with Shiga toxin (Stx) producing Escherichia coli O157:H7 can cause the potentially fatal complication hemolytic uremic syndrome, and currently only supportive therapy is available. Lack of suitable animal models has hindered study of this disease. Induced human intestinal organoids (iHIOs), generated by in vitro differentiation of pluripotent stem cells, represent differentiated human intestinal tissue. We show that iHIOs with addition of human neutrophils can model E. coli intestinal infection and innate cellular responses. Commensal and O157:H7 introduced into the iHIO lumen replicated rapidly achieving high numbers. Commensal E. coli did not cause damage, and were completely contained within the lumen, suggesting defenses, such as mucus production, can constrain non-pathogenic strains. Some O157:H7 initially co-localized with cellular actin. Loss of actin and epithelial integrity was observed after 4 hours. O157:H7 grew as filaments, consistent with activation of the bacterial SOS stress response. SOS is induced by reactive oxygen species (ROS), and O157:H7 infection increased ROS production. Transcriptional profiling (RNAseq) demonstrated that both commensal and O157:H7 upregulated genes associated with gastrointestinal maturation, while infection with O157:H7 upregulated inflammatory responses, including interleukin 8 (IL-8). IL-8 is associated with neutrophil recruitment, and infection with O157:H7 resulted in recruitment of human neutrophils into the iHIO tissue.


The Differential Effects of Insurance Mandates on Health Care Spending for Children's Autism Spectrum Disorder.
There is substantial variation in treatment intensity among children with autism spectrum disorder (ASD). This study asks whether policies that target health care utilization for ASD affect children differentially based on this variation. Specifically, we examine the impact of state-level insurance mandates that require commercial insurers to cover certain treatments for ASD for any fully-insured plan. Using insurance claims between 2008 and 2012 from three national insurers, we used a difference-in-differences approach to compare children with ASD who were subject to mandates to children with ASD who were not. To allow for differential effects, we estimated quantile regressions that evaluate the impact of mandates across the spending distributions of three outcomes: (1) monthly spending on ASD-specific outpatient services; (2) monthly spending on ASD-specific inpatient services; and (3) quarterly spending on psychotropic medications. The change in spending on ASD-specific outpatient services attributable to mandates varied based on the child's level of spending. For those children with ASD who were subject to the mandate, monthly spending for a child in the 95th percentile of the ASD-specific outpatient spending distribution increased by $1460 (P<0.001). In contrast, the effect was only $2 per month for a child in the fifth percentile (P<0.001). Mandates did not significantly affect spending on ASD-specific inpatient services or psychotropic medications. State-level insurance mandates have larger effects for those children with higher levels of spending. To the extent that spending approximates treatment intensity and the underlying severity of ASD, these results suggest that mandates target children with greater service needs.


Headache and blood pressure in primary health care setting in Kuwait.
To identify the types of headache among patients, presenting at a primary health care setting, and to determine the correlation between headache and high blood pressure (BP). This was a cross-sectional study administered in 2 primary health care centers (Fintas and AbuHalifa) of a regional health district in Kuwait from March 2003 to June 2003. Our study included 290 patients with different types of headache, and requested for BP measurement. The sample included patients, in the age group 15-80 years with an overall mean age of 41 (+13 SD) years. The male to female ratio was 1:2.7 and the majority (85%) of patients were Kuwaitis. Tension headache was the most frequent diagnosis (61.7%) among patients presenting with headache, followed by migraine (11.7%). The normal systolic BP was <140 mm Hg (52.8%), while 59% had normal diastolic BP, below 90 mm Hg. Increasing age was found to be associated with high BP levels (p<0.001). Diastolic BP was found significantly higher among males (p<0.01) Tension headache had higher levels of BP while migraine was found to have low levels. None of the sinusitis headache patients had high BP. Positive history of hypertension was found associated with high systolic and diastolic BP in tension and other type of headache (p<0.001). Headache is a very common symptom among patients presenting at the general practice health care setting, with most cases diagnosed with tension-type headache, and females out numbering males. There was no positive correlation between headache and high BP. Physicians should discourage their patients from considering headache as a measure of their BP status.


Latent trajectory studies: the basics, how to interpret the results, and what to report.
In statistics, tools have been developed to estimate individual change over time. Also, the existence of latent trajectories, where individuals are captured by trajectories that are unobserved (latent), can be evaluated (Muthén & Muthén, 2000). The method used to evaluate such trajectories is called Latent Growth Mixture Modeling (LGMM) or Latent Class Growth Modeling (LCGA). The difference between the two models is whether variance within latent classes is allowed for (Jung & Wickrama, 2008). The default approach most often used when estimating such models begins with estimating a single cluster model, where only a single underlying group is presumed. Next, several additional models are estimated with an increasing number of clusters (latent groups or classes). For each of these models, the software is allowed to estimate all parameters without any restrictions. A final model is chosen based on model comparison tools, for example, using the BIC, the bootstrapped chi-square test, or the Lo-Mendell-Rubin test. To ease the use of LGMM/LCGA step by step in this symposium (Van de Schoot, 2015) guidelines are presented which can be used for researchers applying the methods to longitudinal data, for example, the development of posttraumatic stress disorder (PTSD) after trauma (Depaoli, van de Schoot, van Loey, & Sijbrandij, 2015; Galatzer-Levy, 2015). The guidelines include how to use the software Mplus (Muthén & Muthén, 1998-2012) to run the set of models needed to answer the research question: how many latent classes exist in the data? The next step described in the guidelines is how to add covariates/predictors to predict class membership using the three-step approach (Vermunt, 2010). Lastly, it described what essentials to report in the paper. When applying LGMM/LCGA models for the first time, the guidelines presented can be used to guide what models to run and what to report.


Occurrence of Soybean Stem Canker (Diaporthe phaseolorum var. meridionalis) in Wisconsin.
Soybean (Glycine max) developed symptoms characteristic of stem canker during the 2000 to 2003 growing seasons in Wisconsin. Symptoms were widespread in 2003 and were associated with yield losses of ≈1% statewide and as much as 25% in individual fields. Affected plants expressed dieback of foliage beginning at growth stage R3 and progressed until the R6 growth stage. Dark brown lesions were frequently observed at a single node on the lower portion of stems of plants expressing foliage dieback. Fungi were isolated from symptomatic plants collected from seven growers' fields in Rock, Sauk, Veron, and Walworth counties and the Arlington and Marshfield Agricultural Research Stations. Stems with lesions were cut into ≈5-mm pieces, surface disinfested with a 0.5% NaOCl solution for 3 min, rinsed three times in sterile distilled water, and placed on water agar (WA) or potato dextrose agar (PDA) at pH 4.5. Hy-phal tips from colonies of interest were excised and placed on acidified PDA at 25°C under continuous light for 25 to 30 days. In addition to Diaporthe phaseolorum var. caulivora (the cause of northern stem canker), four isolates of D. phaseolorum var. meridionalis (the cause of southern stem canker) were isolated. Colonies of D. phaseolorum var. meridionalis isolates were white, lanose, and became tan with age as previously described for D. phaseolorum var. meridionalis (1). Pycnidia with alpha conidia (no beta conidia) and perithecia with 3.1 to 3.4 × 9.5 to 9.8 μm ascospores formed on oat flakes on acidified WA after 30 days. Stromata were brown to black and irregularly shaped. Four isolates of D. phaseolorum var. meridionalis were tested for pathogenicity in a controlled environment using a cut stem inoculation method (2). Stems of 3-week-old seedlings of cv. Sturdy were cut at the midpoint between the second and third node, and a PDA mycelial plug (4 mm in diameter) was placed on the surface of the cut stems. This method was used to inoculate 15 plants in three replicates for each isolate tested. Inoculated plants were placed in a mist chamber in the dark at 25°C for 4 days and later moved to a greenhouse with a 16-h photoperiod at 24 ± 3°C for 3 days. All plants challenged by this method exhibited stem lesions that were 2 to 3 cm long and of similar color to lesions observed in field-grown plants. For each isolate tested, D. phaseolorum var. meridionalis was reisolated from three randomly selected symptomatic plants. Negative controls with a PDA plug did not produce lesions. To our knowledge, this is the first report of D. phaseolorum var. meridionalis on soybean in Wisconsin. The significance of this report relates to the potential spread of D. phaseolorum var. meridionalis beyond its known southern range in the United States. References: (1) F. A. Fernandez et al. Stem Canker. Pages 33-35 in: Compendium of Soybean Diseases, 4th ed. G. L. Hartman et al., eds. The American Phytopathological Society, St. Paul, MN, 1999. (2) S. Li et al. Plant Dis. 85:1031, 2001.


Power Factor of One Molecule Thick Films and Length Dependence.
There is a rapidly increasing interest in organic thin film thermoelectrics. However, the power factor of one molecule thick organic film, the self-assembled monolayer (SAM), has not yet been determined. This study describes the experimental determination of the power factor in SAMs and its length dependence at an atomic level. As a proof-of-concept, SAMs composed of n-alkanethiolates and oligophenylenethiolates of different lengths are focused. These SAMs were electrically and thermoelectrically characterized on an identical junction platform using a liquid metal top-electrode, allowing the straightforward estimation of the power factor of the monolayers. The results show that the power factor of the alkyl SAMs ranged from 2.0 × 10-8 to 8.0 × 10-12 μW m-1 K-2 and exhibited significant negative length dependence, whereas the conductivity and thermopower of the conjugated SAMs are the two opposing factors that balance the power factor upon an increase in molecular length, exhibiting a maximum power factor of 3.6 × 10-8 μW m-1 K-2. Once correction factors about the ratio of effective contact area to geometrical contact area are considered, the values of power factors can be increased by several orders of magnitude. With a newly derived parametric semiempirical model describing the length dependence of the power factor, it is investigated that one molecule thick films thinner than 10 nm composed of thiophene units can yield power factors rivaling those of famed organic thermoelectric materials based on poly(3,4-ethylenedioxythiophene)/polystyrenesulfonate (PEDOT/PSS) and polyaniline/graphene/double-walled carbon nanotube. Furthermore, how the transition of the transport regime from tunneling to hopping as molecules become long affects power factors is examined.


Intercropped silviculture systems, a key to achieving soil fungal community management in eucalyptus plantations.
Fungi are ubiquitous and important contributors to soil nutrient cycling, playing a vital role in C, N and P turnover, with many fungi having direct beneficial relationships with plants. However, the factors that modulate the soil fungal community are poorly understood. We studied the degree to which the composition of tree species affected the soil fungal community structure and diversity by pyrosequencing the 28S rRNA gene in soil DNA. We were also interested in whether intercropping (mixed plantation of two plant species) could be used to select fungal species. More than 50,000 high quality sequences were analyzed from three treatments: monoculture of Eucalyptus; monoculture of Acacia mangium; and a mixed plantation with both species sampled 2 and 3 years after planting. We found that the plant type had a major effect on the soil fungal community structure, with 75% of the sequences from the Eucalyptus soil belonging to Basidiomycota and 19% to Ascomycota, and the Acacia soil having a sequence distribution of 28% and 62%, respectively. The intercropping of Acacia mangium in a Eucalyptus plantation significantly increased the number of fungal genera and the diversity indices and introduced or increased the frequency of several genera that were not found in the monoculture cultivation samples. Our results suggest that management of soil fungi is possible by manipulating the composition of the plant community, and intercropped systems can be a means to achieve that.


Patient-Centered Outcomes for Skin Cancer Management: Utilization of a Patient Delphi Process to Identify Important Treatment Themes.
Patient-reported outcomes are critical to research directed at maximizing patient benefit. The outcomes patients consider most relevant in the treatment of skin cancer have not been directly investigated. To develop a ranked list of outcomes deemed most important by patients with skin cancer through a proctored Delphi process. Twenty-one patients with a history of skin cancer volunteered to participate in the patient summit. The patient members participated in a guided discussion to generate a long list of potentially relevant outcomes. The list was then condensed and ranked through 2 rounds of a proctored Delphi process. Patients were diverse in their skin cancer histories and complexities, ages, and states of residence. Twelve themes were rated as highly important by 70% or more of participants. Most of these themes related to patient education and the collaborative nature of the physician-patient relationship. Fear of recurrence and cosmetic outcome were also highly rated. Limitations include a preponderance of older patients and patients from the midwest and northeast. Patients with skin cancer overwhelmingly prioritize a shared decision-making process, in which they are actively engaged and value detailed education regarding their disease. This should inform future research directed at skin cancer treatment and current physician-patient interactions.


Effects of psychiatric disorders and psychotropic medications on prolactin and bone metabolism.
Osteoporosis occurs in common psychiatric conditions and causes significant morbidity. Many neuroleptic medications can cause hyperprolactinemia, which can then potentially be associated with bone loss. Few reviews have thus far addressed this issue. We have consolidated information from studies that examined effects of psychiatric conditions and their treatment on bone metabolism. We searched PubMed for original articles and reviews published between 1976 and 2004 that described changes in bone metabolism in psychiatric disorders and examined prolactin elevations with neuroleptic medications. Keywords used were major depressive disorder, bipolar disorder, schizophrenia, bone density, bone metabolism, hyperprolactinemia, typical antipsychotics, and atypical antipsychotics. 160 articles published in peer-reviewed journals were identified and are summarized, with greater emphasis given to data from larger, controlled studies. Schizophrenia and major mood disorders are often associated with perturbations in bone metabolism related to factors including nutritional alterations, smoking, and hypogonadotropic hypogonadism, with or without medication-induced hyperprolactinemia. Polydipsia can contribute to bone loss in schizophrenia, whereas hypercortisolemia is often associated with low bone density in depression. Lithium in bipolar disorder and thyroid-stimulating hormone-suppressive doses of L-thyroxine have a negative impact on bone health. Mood stabilizers such as carbamazepine and valproate can also affect bone density. Hyperprolactinemia may lead to bone loss only if associated with untreated amenorrhea in women or testosterone deficiency in men. Some atypical neuroleptics, by causing lesser elevations in prolactin, may therefore have a less marked impact on bone than typical neuroleptics. Because significant morbidity is associated with low bone density and many psychiatric conditions may have a negative impact on bone metabolism, bone density evaluation should be considered an integral component of chronic medical care of these disorders, and risk factors should be identified and addressed.


Diagnostic Performance of Conventional X-ray for Detecting Foreign Bodies in the Upper Digestive Tract: A Systematic Review and Diagnostic Meta-Analysis.
Foreign body (FB) ingestion is a common clinical problem in acute settings. Detecting FBs in the upper digestive tract is challenging. The conventional X-ray is usually the first-line imaging tool to detect FBs. However, its diagnostic performance is inconsistent in the literature. In this study, we performed a systematic review and meta-analysis to determine the diagnostic performance of the conventional X-ray for detecting FBs in the upper digestive tract. We conducted a systematic search of PubMed, Embase, Cochrane Library, Web of Science, and Scopus until 1 August 2020. Prospective or retrospective studies investigating the diagnostic accuracy of conventional X-rays for detecting FBs in the upper digestive tract in patients of all ages were included. The Quality Assessment of Studies of Diagnostic Accuracy-2 tool was used to review the quality of included studies. We used a bivariate random-effects model to calculate diagnostic accuracy parameters. Heterogeneity was assessed using I2 statistics. We included 17 studies (n = 4809) in the meta-analysis. Of the 17 studies, most studies were rated as having a high risk of bias. Conventional X-rays had a pooled sensitivity of 0.58 (95% confidence interval [CI] = 0.36-0.77, I2 = 98.52) and a pooled specificity of 0.94 (95% CI = 0.87-0.98, I2 = 94.49) for detecting FBs in the upper digestive tract. The heterogeneity was considerable. The area under the summary receiver operating characteristic curve was 0.91 (95% CI = 0.88-0.93). Deek's funnel plot asymmetry test results revealed no significant publication bias (p = 0.41). The overall sensitivity and specificity of conventional X-rays were low and high, respectively, for detecting FBs in the upper digestive tract. Hence, conventional X-rays to exclude patients without upper FBs in the digestive tract are not recommended. Further imaging or endoscopic examinations should be performed for at-risk patients.


When emotions run high: A critical role for context in the unfolding of dynamic, real-life facial affect.
Although positive and negative affect are assumed to be highly distinct, recent work has shown that facial valence of positive and negative situations may be highly confusable, especially when the emotions are intense. However, previous work has relied exclusively on static images, portraying a single peak frame of the emotional display. Dynamic expressions, on the other hand, convey a far broader representation of the emotional reaction, but are they diagnostic of the situational valence? Participants (N = 245) watched videos portraying reactions to real-life highly positive situations and evaluated the affective valence of the target. Video information was controlled by: (a) truncating the movies after 5, 10, or 20 seconds from the start, and by (b) digitally manipulating the videos such that only the face was visible with no context, only the context was visible with no face, or the face appeared in context. Results indicate that during real-life intense positive situations, facial expressions alone were rated as negative and failed to convey diagnostic information about the positive situational valence even at the most extended presentation durations. By contrast, when contextual information appeared alone or with the face, participants accurately rated the target as feeling positive, and this positivity increased with extended viewing duration. These findings suggests poor coupling between facial valence and felt emotions, supporting the notion that when emotions run high, the diagnostic power of facial expressions is reduced. Conversely, the findings demonstrate an inherent role for contextual information in the recognition of real-life intense faces. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Effective and new potent drug combination: Histone deacetylase and Wnt/β-catenin pathway inhibitors in lung carcinoma cells.
Lung cancer is the most commonly diagnosed cancer worldwide with a high mortality rate. In this study, the therapeutic effect of combination valproic acid and niclosamide was investigated on human lung cancer cell line. The effects of the compounds alone and combination therapy on cell viability were determined by sulforhodamine B and adenosine 5'-triphosphate viability assays. Flow cytometry was used to determine the cell death mechanism and DNA damage levels responsible for the cytotoxic effects of combination therapy. The presence of apoptosis in cells was supported by fluorescence microscopy and also by using inhibitors of the apoptotic signaling pathway. The increase in cellular reactive oxygen species (ROS) level in combination therapy was determined by H2DCFDA staining. The effect of N-acetyl-l-cysteine combination on ROS increase was investigated on cell viability. In addition, the expression levels of the proteins associated with epigenetic regulation and cell death were analyzed by Western blotting and gene expression levels were determined using real-time quantitative polymerase chain reaction.It was observed that the combination therapy showed a cytotoxic effect on the A549 lung cancer cells compared to the individual use of the inhibitors. The absence of this effect on normal lung cells revealed the presence of a selective toxic effect. When the mechanism of cytotoxicity is examined, it has been observed that combination therapy initiates the activation of tumor necrosis receptors and causes apoptosis by activated caspase. It was also observed that this extrinsic apoptotic pathway was activated on the mitochondrial pathway. In addition, ER stress and mitochondrial membrane potential loss associated with increased ROS levels induce cell death. When the data in this study were evaluated, combination therapy caused a dramatic decrease in cell viability by inducing the extrinsic apoptotic pathway in lung cancer cell line. Therefore, it was concluded that it can be used as an effective and new treatment option for lung cancer.


Insertion-and-deletion-derived tumour-specific neoantigens and the immunogenic phenotype: a pan-cancer analysis.
The focus of tumour-specific antigen analyses has been on single nucleotide variants (SNVs), with the contribution of small insertions and deletions (indels) less well characterised. We investigated whether the frameshift nature of indel mutations, which create novel open reading frames and a large quantity of mutagenic peptides highly distinct from self, might contribute to the immunogenic phenotype. We analysed whole-exome sequencing data from 5777 solid tumours, spanning 19 cancer types from The Cancer Genome Atlas. We compared the proportion and number of indels across the cohort, with a subset of results replicated in two independent datasets. We assessed in-silico tumour-specific neoantigen predictions by mutation type with pan-cancer analysis, together with RNAseq profiling in renal clear cell carcinoma cases (n=392), to compare immune gene expression across patient subgroups. Associations between indel burden and treatment response were assessed across four checkpoint inhibitor datasets. We observed renal cell carcinomas to have the highest proportion (0·12) and number of indel mutations across the pan-cancer cohort (p<2·2 × 10-16), more than double the median proportion of indel mutations in all other cancer types examined. Analysis of tumour-specific neoantigens showed that enrichment of indel mutations for high-affinity binders was three times that of non-synonymous SNV mutations. Furthermore, neoantigens derived from indel mutations were nine times enriched for mutant specific binding, as compared with non-synonymous SNV derived neoantigens. Immune gene expression analysis in the renal clear cell carcinoma cohort showed that the presence of mutant-specific neoantigens was associated with upregulation of antigen presentation genes, which correlated (r=0·78) with T-cell activation as measured by CD8-positive expression. Finally, analysis of checkpoint inhibitor response data revealed frameshift indel count to be significantly associated with checkpoint inhibitor response across three separate melanoma cohorts (p=4·7 × 10-4). Renal cell carcinomas have the highest pan-cancer proportion and number of indel mutations. Evidence suggests indels are a highly immunogenic mutational class, which can trigger an increased abundance of neoantigens and greater mutant-binding specificity. Cancer Research UK, UK National Institute for Health Research (NIHR) at the Royal Marsden Hospital National Health Service Foundation Trust, Institute of Cancer Research and University College London Hospitals Biomedical Research Centres, the UK Medical Research Council, the Rosetrees Trust, Novo Nordisk Foundation, the Prostate Cancer Foundation, the Breast Cancer Research Foundation, the European Research Council.


Role of interleukin 6 for differential responsiveness of naive and memory CD4+ T cells in CD2-mediated activation.
The present study was undertaken to elucidate different requirements for CD2-mediated activation of naive (CD45RO-) and memory (CD45RO+) CD4+ T cells. A mitogenic combination of anti-CD2 (anti-T11(2) and anti-T11(3] mAbs could effectively induce the proliferation of memory CD4+ T cells even in the absence of monocytes. In marked contrast, naive CD4+ T cells did not disclose any proliferative responses to anti-CD2 mAbs, when monocytes were absent in culture. This differential responsiveness of naive and memory CD4+ T cells appeared to be related largely to a difference in IL-6-producing ability between both populations. IL-6 among monocyte-derived cytokines could correct unresponsiveness of naive CD4+ T cells to anti-CD2 stimulation. Unlike naive CD4+ T cells, memory CD4+ T cells produced IL-6 by themselves, with its mRNA being expressed on anti-CD2 stimulation. Anti-IL-6R mAb significantly inhibited proliferation of memory CD4+ T cells seen in the anti-CD2-stimulated cultures without monocytes, indicating the involvement of their own production of IL-6 in CD2-mediated activation. The results suggest an essential role of IL-6 for triggering of CD4+ T cells via the CD2 molecule.


Extensive loss of cell-cycle and DNA repair genes in an ancient lineage of bipolar budding yeasts.
Cell-cycle checkpoints and DNA repair processes protect organisms from potentially lethal mutational damage. Compared to other budding yeasts in the subphylum Saccharomycotina, we noticed that a lineage in the genus Hanseniaspora exhibited very high evolutionary rates, low Guanine-Cytosine (GC) content, small genome sizes, and lower gene numbers. To better understand Hanseniaspora evolution, we analyzed 25 genomes, including 11 newly sequenced, representing 18/21 known species in the genus. Our phylogenomic analyses identify two Hanseniaspora lineages, a faster-evolving lineage (FEL), which began diversifying approximately 87 million years ago (mya), and a slower-evolving lineage (SEL), which began diversifying approximately 54 mya. Remarkably, both lineages lost genes associated with the cell cycle and genome integrity, but these losses were greater in the FEL. E.g., all species lost the cell-cycle regulator WHIskey 5 (WHI5), and the FEL lost components of the spindle checkpoint pathway (e.g., Mitotic Arrest-Deficient 1 [MAD1], Mitotic Arrest-Deficient 2 [MAD2]) and DNA-damage-checkpoint pathway (e.g., Mitosis Entry Checkpoint 3 [MEC3], RADiation sensitive 9 [RAD9]). Similarly, both lineages lost genes involved in DNA repair pathways, including the DNA glycosylase gene 3-MethylAdenine DNA Glycosylase 1 (MAG1), which is part of the base-excision repair pathway, and the DNA photolyase gene PHotoreactivation Repair deficient 1 (PHR1), which is involved in pyrimidine dimer repair. Strikingly, the FEL lost 33 additional genes, including polymerases (i.e., POLymerase 4 [POL4] and POL32) and telomere-associated genes (e.g., Repressor/activator site binding protein-Interacting Factor 1 [RIF1], Replication Factor A 3 [RFA3], Cell Division Cycle 13 [CDC13], Pbp1p Binding Protein [PBP2]). Echoing these losses, molecular evolutionary analyses reveal that, compared to the SEL, the FEL stem lineage underwent a burst of accelerated evolution, which resulted in greater mutational loads, homopolymer instabilities, and higher fractions of mutations associated with the common endogenously damaged base, 8-oxoguanine. We conclude that Hanseniaspora is an ancient lineage that has diversified and thrived, despite lacking many otherwise highly conserved cell-cycle and genome integrity genes and pathways, and may represent a novel, to our knowledge, system for studying cellular life without them.


A point mutation in the hair cell nicotinic cholinergic receptor prolongs cochlear inhibition and enhances noise protection.
The transduction of sound in the auditory periphery, the cochlea, is inhibited by efferent cholinergic neurons projecting from the brainstem and synapsing directly on mechanosensory hair cells. One fundamental question in auditory neuroscience is what role(s) this feedback plays in our ability to hear. In the present study, we have engineered a genetically modified mouse model in which the magnitude and duration of efferent cholinergic effects are increased, and we assess the consequences of this manipulation on cochlear function. We generated the Chrna9L9'T line of knockin mice with a threonine for leucine change (L9'T) at position 9' of the second transmembrane domain of the alpha9 nicotinic cholinergic subunit, rendering alpha9-containing receptors that were hypersensitive to acetylcholine and had slower desensitization kinetics. The Chrna9L9'T allele produced a 3-fold prolongation of efferent synaptic currents in vitro. In vivo, Chrna9L9'T mice had baseline elevation of cochlear thresholds and efferent-mediated inhibition of cochlear responses was dramatically enhanced and lengthened: both effects were reversed by strychnine blockade of the alpha9alpha10 hair cell nicotinic receptor. Importantly, relative to their wild-type littermates, Chrna9(L9'T/L9'T) mice showed less permanent hearing loss following exposure to intense noise. Thus, a point mutation designed to alter alpha9alpha10 receptor gating has provided an animal model in which not only is efferent inhibition more powerful, but also one in which sound-induced hearing loss can be restrained, indicating the ability of efferent feedback to ameliorate sound trauma.


Bad oscillations in Parkinson's disease.
Recordings in humans as a result of functional neurosurgery have revealed a tendency for basal ganglia neurons to oscillate and synchronise their activity, giving rise to a rhythmic population activity, manifest as oscillatory local field potentials. The most important activity is synchronised oscillation in the beta band (13-30 Hz), which has been picked up at various sites within the basal ganglia-cortical loop in PD. Dopaminergic medication and movement suppress this activity, with the timing and degree of suppression closely correlating with behavioural performance. Accordingly synchronisation in the beta band has been hypothesised to be essentially antikinetic in nature and pathophysiologically relevant to bradykinesia.


Baseline survey of the anatomical microbial ecology of an important food plant: Solanum lycopersicum (tomato).
Research to understand and control microbiological risks associated with the consumption of fresh fruits and vegetables has examined many environments in the farm to fork continuum. An important data gap however, that remains poorly studied is the baseline description of microflora that may be associated with plant anatomy either endemically or in response to environmental pressures. Specific anatomical niches of plants may contribute to persistence of human pathogens in agricultural environments in ways we have yet to describe. Tomatoes have been implicated in outbreaks of Salmonella at least 17 times during the years spanning 1990 to 2010. Our research seeks to provide a baseline description of the tomato microbiome and possibly identify whether or not there is something distinctive about tomatoes or their growing ecology that contributes to persistence of Salmonella in this important food crop. DNA was recovered from washes of epiphytic surfaces of tomato anatomical organs; leaves, stems, roots, flowers and fruits of Solanum lycopersicum (BHN602), grown at a site in close proximity to commercial farms previously implicated in tomato-Salmonella outbreaks. DNA was amplified for targeted 16S and 18S rRNA genes and sheared for shotgun metagenomic sequencing. Amplicons and metagenomes were used to describe "native" bacterial microflora for diverse anatomical parts of Virginia-grown tomatoes. Distinct groupings of microbial communities were associated with different tomato plant organs and a gradient of compositional similarity could be correlated to the distance of a given plant part from the soil. Unique bacterial phylotypes (at 95% identity) were associated with fruits and flowers of tomato plants. These include Microvirga, Pseudomonas, Sphingomonas, Brachybacterium, Rhizobiales, Paracocccus, Chryseomonas and Microbacterium. The most frequently observed bacterial taxa across aerial plant regions were Pseudomonas and Xanthomonas. Dominant fungal taxa that could be identified to genus with 18S amplicons included Hypocrea, Aureobasidium and Cryptococcus. No definitive presence of Salmonella could be confirmed in any of the plant samples, although 16S sequences suggested that closely related genera were present on leaves, fruits and roots.


Potency ranking of triterpenoids as inducers of a cytoprotective enzyme and as inhibitors of a cellular inflammatory response via their electron affinity and their electrophilicity index.
Electron affinity (EA) and electrophilicity index (omega) of 16 synthetic triterpenoids (TP), previously identified as inducers of cytoprotective enzymes and as inhibitors of cellular inflammatory responses, have been calculated by the molecular orbital method. Linear correlations were obtained by plotting the values of EA, as well as those of omega versus (i) the potencies of induction of NAD(P)H quinone reductase (NQO1, EC 220.127.116.11), a cytoprotective enzyme, expressed via the concentration of TP required to double the specific activity of NQO1 (CD value) and (ii) the values of their anti-inflammatory activity expressed via the IC-50 of TP for suppression of upregulation of inducible nitric oxide synthase (iNOS, EC 126.96.36.199), both previously experimentally determined. The observed correlations demonstrate quantitatively for a series of triterpenoids that their electrophilicity is a major factor determining their potency as inducers of the cytoprotective phase 2 response and as inhibitors of inflammatory processes.


The eSMART study protocol: a randomised controlled trial to evaluate electronic symptom management using the advanced symptom management system (ASyMS) remote technology for patients with cancer.
While some evidence exists that real-time remote symptom monitoring devices can decrease morbidity and prevent unplanned admissions in oncology patients, overall, these studies have significant methodological weaknesses. The electronic Symptom Management using the Advanced Symptom Management System (ASyMS) Remote Technology (eSMART) study is designed to specifically address these weaknesses with an appropriately powered, repeated-measures, parallel-group stratified randomised controlled trial of oncology patients. A total of 1108 patients scheduled to commence first-line chemotherapy (CTX) for breast, colorectal or haematological cancer will be recruited from multiple sites across five European countries.Patients will be randomised (1:1) to the ASyMS intervention (intervention group) or to standard care currently available at each site (control group). Patients in the control and intervention groups will complete a demographic and clinical questionnaire, as well as a set of valid and reliable electronic patient-reported outcome measures at enrolment, after each of their CTX cycles (up to a maximum of six cycles) and at 3, 6, 9 and 12 months after completion of their sixth cycle of CTX. Outcomes that will be assessed include symptom burden (primary outcome), quality of life, supportive care needs, anxiety, self-care self-efficacy, work limitations and cost effectiveness and, from a health professional perspective, changes in clinical practice (secondary outcomes). Ethical approval will be obtained prior to the implementation of all major study amendments. Applications will be submitted to all of the ethics committees that granted initial approval.eSMART received approval from the relevant ethics committees at all of the clinical sites across the five participating countries. In collaboration with the European Cancer Patient Coalition (ECPC), the trial results will be disseminated through publications in scientific journals, presentations at international conferences, and postings on the eSMART website and other relevant clinician and consumer websites; establishment of an eSMART website (www.esmartproject.eu) with publicly accessible general information; creation of an eSMART Twitter Handle, and production of a toolkit for implementing/utilising the ASyMS technology in a variety of clinical practices and other transferable health care contexts. NCT02356081.


Serotonin precursors in chronic primary headache. A double-blind cross-over study with L-5-hydroxytryptophan vs. placebo.
Serotonin (5-HT) plays a crucial role in mediating the descending pain inhibitory systems and in the pathophysiology of migraine. Previous studies regarding the use of 5-Hydroxytryptophan (5-HTP), the active precursor of 5-HT, in the treatment of Chronic Primary Headache (CPH) have been inconclusive so far. In order to assess the efficacy of the serotonin active precursor in chronic headache prophylaxis, a double-blind cross-over study has been carried out in 31 patients with CPH, comparing L-5-HTP to placebo. Clinical syndromes included: (a) migraine (16 patients); (b) mixed headache (6 patients); (c) psychogenic headache (5 patients); (d) muscle contraction headache (4 patients). L-5-HTP was administered for two months at daily doses of 400 mg p.o. The reduction in severity and frequency of headache in patients taking the active drug and placebo was noted. Mood patterns were also taken into consideration. L-5-HTP proved to be more effective than placebo in reducing both headache frequency and severity, but the difference was not statistically significant. Favourable responses (greater than 50% average reduction in headache symptoms) were obtained in 48% of the cases after the second month of treatment. No significant difference in therapeutic response was observed as related to different clinical syndromes, except for psychogenic headache patients, who responded poorly to the active drug. Side effects, experienced in 19% of the cases, were generally mild and transient. We conclude that L-5-HTP is a medication of moderate efficacy and remarkable safety, providing us with another alternative approach to CPH prophylaxis.


Aspherics and surface generation; curve generation on a blanchard.
Techniques used to produce aspheric and other irregular curves are described in this article. Although the machinery is not unique, the manner in which it is applied is felt to be unusual. Typical of the many challenging jobs this company has produced was the grinding and polishing of numerous massive radiationshielding windows. These windows had a density of 3.2. Their physical dimensions were 2.1 m by 2.6 m by 125 mm thick. The weight of these windows was 1915 kg. A description of the equipment and techniques used will be given in a future article.


Fenestrated endovascular aneurysm repair is associated with lower perioperative morbidity and mortality compared with open repair for complex abdominal aortic aneurysms.
The Zenith Fenestrated Endovascular Graft (ZFEN; Cook Medical, Bloomington, Ind) has expanded the anatomic eligibility of endovascular aneurysm repair (EVAR) for complex abdominal aortic aneurysms (AAAs). Current data on ZFEN mainly consist of single-institution experiences and show conflicting results. Therefore, we compared perioperative outcomes after repair using ZFEN with open complex AAA repair and infrarenal EVAR in a nationwide multicenter registry. We identified all patients undergoing elective AAA repair using ZFEN, open complex AAA repair, and standard infrarenal EVAR between 2012 and 2016 within the American College of Surgeons National Surgical Quality Improvement Program targeted vascular module. Open complex AAA repairs were defined as those with a juxtarenal or suprarenal proximal AAA extent in combination with an aortic cross-clamping position that was above at least one renal artery. The primary outcome was perioperative mortality, defined as death within 30 days or within the index hospitalization. Secondary outcomes included postoperative renal dysfunction (creatinine concentration increase of >2 mg/dL from preoperative value or new dialysis), occurrence of any complication, procedure times, blood transfusion rates, and length of stay. To account for baseline differences, we calculated propensity scores and employed inverse probability-weighted logistic regression. We identified 6825 AAA repairs-220 ZFENs, 181 open complex AAA repairs, and 6424 infrarenal EVARs. Univariate analysis of ZFEN compared with open complex AAA repair demonstrated lower rates of perioperative mortality (1.8% vs 8.8%; P = .001), postoperative renal dysfunction (1.4% vs 7.7%; P = .002), and overall complications (11% vs 33%; P < .001). In addition, fewer patients undergoing ZFEN received blood transfusions (22% vs 73%; P < .001), and median length of stay was shorter (2 vs 7 days; P < .001). After adjustment, open complex AAA repair was associated with higher odds of perioperative mortality (odds ratio [OR], 4.9; 95% confidence interval [CI], 1.4-18), postoperative renal dysfunction (OR, 13; 95% CI, 3.6-49), and overall complication rates (OR, 4.2; 95% CI, 2.3-7.5) compared with ZFEN. Compared with infrarenal EVAR, ZFEN presented comparable rates of perioperative mortality (1.8% vs 0.8%; P = .084), renal dysfunction (1.4% vs 0.7%; P = .19), and any complication (11% vs 7.7%; P = .09). Furthermore, after adjustment, there was no significant difference between the odds of perioperative mortality, postoperative renal dysfunction, or any complication between infrarenal EVAR and ZFEN. ZFEN is associated with lower perioperative morbidity and mortality compared with open complex AAA repair, and outcomes are comparable to those of infrarenal EVAR. Long-term durability of ZFEN compared with open complex AAA repair warrants future research.


Two-dimensional gap analysis: a tool for efficient conservation planning and biodiversity policy implementation.
The maintenance of biodiversity by securing representative and well-connected habitat networks in managed landscapes requires a wise combination of protection, management, and restoration of habitats at several scales. We suggest that the integration of natural and social sciences in the form of "Two-dimensional gap analysis" is an efficient tool for the implementation of biodiversity policies. The tool links biologically relevant "horizontal" ecological issues with "vertical" issues related to institutions and other societal issues. Using forest biodiversity as an example, we illustrate how one can combine ecological and institutional aspects of biodiversity conservation, thus facilitating environmentally sustainable regional development. In particular, we use regional gap analysis for identification of focal forest types, habitat modelling for ascertaining the functional connectivity of "green infrastructures", as tools for the horizontal gap analysis. For the vertical dimension we suggest how the social sciences can be used for assessing the success in the implementation of biodiversity policies in real landscapes by identifying institutional obstacles while implementing policies. We argue that this interdisciplinary approach could be applied in a whole range of other environments including other terrestrial biota and aquatic ecosystems where functional habitat connectivity, nonlinear response to habitat loss and a multitude of economic and social interests co-occur in the same landscape.


Prostaglandin D2 regulates CD4+ memory T cell trafficking across blood vascular endothelium and primes these cells for clearance across lymphatic endothelium.
Memory lymphocytes support inflammatory and immune responses. To do this, they enter tissue via blood vascular endothelial cells (BVEC) and leave tissue via lymphatic vascular endothelial cells (LVEC). In this study, we describe a hierarchy of signals, including novel regulatory steps, which direct the sequential migration of human T cells across the blood and the lymphatic EC. Cytokine-stimulated (TNF and IFN) human BVEC preferentially recruited memory T cells from purified PBL. Lymphocyte recruitment from flow could be blocked using a function-neutralizing Ab against CXCR3. However, a receptor antagonist directed against the PGD(2) receptor DP2 (formerly chemoattractant receptor-homologous molecule expressed on Th2 cells) inhibited transendothelial migration, demonstrating that the sequential delivery of the chemokine and prostanoid signals was required for efficient lymphocyte recruitment. CD4(+) T cells recruited by BVEC migrated with significantly greater efficiency across a second barrier of human LVEC, an effect reproduced by the addition of exogenous PGD(2) to nonmigrated cells. Migration across BVEC or exogenous PGD(2) modified the function, but not the expression, of CCR7, so that chemotaxis toward CCL21 was significantly enhanced. Thus, chemokines may not regulate all stages of lymphocyte migration during inflammation, and paradigms describing their trafficking may need to account for the role of PGD(2).


The recycling of apolipoprotein E in primary cultures of mouse hepatocytes. Evidence for a physiologic connection to high density lipoprotein metabolism.
Internalization of apoE-containing very low density protein (VLDL) by hepatocytes in vivo and in vitro leads to apoE recycling and resecretion. Because of the role of apoE in VLDL metabolism, apoE recycling may influence lipoprotein assembly or remnant uptake. However, apoE is also a HDL protein, and apoE recycling may be related to reverse cholesterol transport. To investigate apoE recycling, apoE(-/-) mouse hepatocytes were incubated (pulsed) with wild-type mouse lipoproteins, and cells and media were collected at chase periods up to 24 h. When cells were pulsed with VLDL, apoE was resecreted within 30 min. Although the mass of apoE in the media decreased with time, it could be detected up to 24 h after the pulse. Intact intracellular apoE was also detectable 24 h after the pulse. ApoE was also resecreted when cells were pulsed with HDL. When apoA-I was included in the chase media after a pulse with VLDL, apoE resecretion increased 4-fold. Furthermore, human apoE was resecreted from wild-type mouse hepatocytes after a pulse with human VLDL. Finally, apoE was resecreted from mouse peritoneal macrophages after pulsing with VLDL. We conclude that 1) HDL apoE recycles in a quantitatively comparable fashion to VLDL apoE; 2) apoE recycling can be modulated by extracellular apoA-I but is not affected by endogenous apoE; and 3) recycling occurs in macrophages as well as in hepatocytes, suggesting that the process is not cell-specific.


Self-assessment in neurological surgery: the SANS wired white paper.
Periodic self-assessment is an important component of postgraduate medical education and certification. The Self-assessment in Neurological Surgery (SANS) examination has a history of usefulness for neurosurgical written and oral board examination preparation. The SANS Wired test represents a recreation of the original SANS test using an Internet-based platform. Advanced functionality and contemporary content have been added to meet the increasing requirements for demonstration of lifelong learning and self-assessment as part of the Maintenance of Certification (MOC) process. A needs assessment was performed before the development of SANS Wired. Postexamination surveys for users obtaining continuing medical education credit were analyzed for user satisfaction with the platform and content. Test-item performance metrics were analyzed based on first-attempt responses. The economic value of SANS Wired as a component of the mandatory MOC process was evaluated. The needs assessment identified a deficiency of available content and mechanisms for neurosurgery-specific self-assessment. As of October 15, 2005, a total of 588 individuals had subscribed to SANS Wired (28 institutional licenses accounted for 229 users). Eighty-eight post-test surveys were analyzed for user satisfaction. Ninety-four percent of responses indicated that learning objectives had been met. Initial performance metrics show an average point-biserial discrimination index of +0.18 for all test items. The SANS Wired system seems to be an effective platform for neurosurgical self-assessment and is acceptable to users. The system satisfies, in part, two of the four required components of MOC, namely, periodic self-assessment and demonstration of cognitive expertise. It is endorsed by the American Board of Neurological Surgeons for MOC.


A novel surface antigen expressed by a subset of human CD3- CD16+ natural killer cells. Role in cell activation and regulation of cytolytic function.
The GL183 mAb was obtained by immunizing BALB/c mice with the E57 clone (CD7+CD2+CD3-CD16+CD56+) derived from human peripheral blood NK cells. In human peripheral blood, GL183-reactive cells ranged between 2 and 12% (mean 6.5%) in 10 different donors. Double fluorescence and FACS analysis showed that GL183+ cells were consistently included in the CD56+ or CD16+ cell populations. Moreover, since only a fraction of CD56+ or CD16+ cells (approximately 40%) coexpressed GL183 surface antigen, reactivity with GL183 mAb appears to define two subsets within the CD3- lymphocyte population expressing NK cell markers. Although, the majority of GL183+ cells were CD3-, approximately 1% expressed CD3 surface antigens. As shown by clonal analysis, these infrequent CD3+GL183+ cells coexpressed CD56 and CD16 antigens. Cloning of CD3-GL183+ or CD3-GL183- cell populations under limiting dilution conditions yielded clonal progenies that maintained their original surface phenotype. Therefore, expression or lack of expression of GL183 surface antigens represents a stable phenotypic property of a subset of human CD3- NK cells. Immunoprecipitation experiments and two-dimensional PAGE analysis indicated that GL183-reactive molecules were represented in different clones either by a single 58-kD chain or, more frequently, by two chains of approximately 55 and approximately 58 kD, respectively. Analysis of GL183+ or GL183- NK clones for their ability to lyse human (IGROV I) or murine (P815) tumor target cells indicated that GL183- clones were, on average, fivefold more efficient in inducing target cell lysis. GL183+ and GL183- clones produced comparable levels of TNF-alpha in response to PHA plus PMA or anti-CD16 mAb plus PMA. Importantly, production of TNF-alpha was also induced by stimulation of GL183+ clones with GL183 mAb plus PMA. These data indicated that GL183 antigen could mediate cell triggering. This concept was confirmed by the analysis of Ca2+ mobilization, as GL183 mAb induced (in GL183+ clones) increments of [Ca2+]i comparable with those induced by PHA. Moreover, GL183 mAb, or its F(ab')2 fragments, strongly enhanced the cytolytic activity of GL183+ clones against a panel of human tumor target cells, including U937, Raji, IGROV I, M14, and A549. In contrast, GL183 mAb, but not the F(ab')2 fragments, sharply inhibited the cytolytic activity of the same clones against P815, M12, and P3U1 murine target cells. In this case, the effect of GL183 mAb (inhibition) was opposite that of PHA or of stimulatory anti-CD2 or anti-CD16 mAbs, which consistently enhanced the target cell lysis.(ABSTRACT TRUNCATED AT 400 WORDS)


Cell proliferation in cancer prevention; effects of preventive agents on estrogen-related endometrial carcinogenesis model and on an in vitro model in human colorectal cells.
Proto-oncogenes such as c-fos, c-jun and c-myc are known to relate to cell proliferation and differentiation. Some oriental herbal medicines like Glycyrrhizae radix or Juzen-taiho-to were found to suppress estradiol-17 beta (E2)-induced expression of c-fos/jun in uterine corpus and inhibited N-methyl-N-nitrosourea and E2-induced endometrial carcinogenesis in mice. It is suggested that the effects of such oriental drugs are exerted probably through suppression of estrogen-induced c-fos/jun expression and they are promising preventing agents for endometrial cancers. In the combined in vitro assay for cell proliferation (MTS assay) and apoptosis (DNA fragmentation) in human colorectal cancer cells (Colo 320), a number of naturally occurring chemopreventive agents such as curcumin, quercetin, auraptene, 1'-acetoxychavicol acetate (ACA) and indole-3-carbinol were shown to generate apoptosis as well as to inhibit cell proliferation. The results suggest a mode of action of these chemopreventive agents and also imply that such in vitro short term assay is useful for detection of new agents for cancer prevention.


Distinguishing uniform switching from filamentary switching in resistance memory using a fracture test.
Resistance random access memory (RRAM) is a rapidly developing emergent nanotechnology. For practical applications and basic understanding, it is important to ascertain whether RRAM undergoes uniform or filamentary switching, but on this point previous area-scaling studies have often shown ambiguous and conflicting findings. Here we demonstrate a simple test-physically breaking the device into two and studying their characteristics individually-can make a definitive determination. Our experiment on two prototypical RRAMs found that one (the nanometallic memory) switches and conducts uniformly while the other (the filamentary memory) does not. It also probes the statistics of nanofilaments: the resistance statistics of the filamentary memory reveals for the first time a large population of partially developed filaments in addition to the filament that dominates switching. Remarkably, the filamentary memory can also be stress-switched to a lower resistance state during fracture, which is reminiscent of stress-switching of the nanometallic memory and may be taken as direct evidence of electron-phonon interaction in the filaments.


Characterisation of fibroblast-like synoviocytes from a murine model of joint inflammation.
Fibroblast-like synoviocytes (FLS) play a central role in defining the stromal environment in inflammatory joint diseases. Despite a growing use of FLS isolated from murine inflammatory models, a detailed characterisation of these cells has not been performed. In this study, FLS were isolated from inflamed joints of mice expressing both the T cell receptor transgene KRN and the MHC class II molecule Ag7 (K/BxN mice) and their purity in culture determined by immunofluorescence and real-time reverse transcription polymerase chain reaction (real-time RT-PCR). Basal expression of proinflammatory genes was determined by real-time RT-PCR. Secreted interleukin 6 (IL-6) was measured by enzyme-linked immunosorbent assay (ELISA), and its regulation by tumor necrosis factor-alpha (TNF-α and corticosterone (the major glucocorticoid in rodents) measured relative to other mesenchymal cell populations. Purity of FLS culture was identified by positive expression of fibronectin, prolyl 4-hydroxylase, cluster of differentiation 90.2 (CD90.2) and 248 (CD248) in greater than 98% of the population. Cultured FLS were able to migrate and invade through matrigel, a process enhanced in the presence of TNF-α. FLS isolated from K/BxN mice possessed significantly greater basal expression of the inflammatory markers IL-6, chemokine ligand 2 (CCL-2) and vascular cell adhesion molecule 1 (VCAM-1) when compared to FLS isolated from non-inflamed tissue (IL-6, 3.6 fold; CCL-2, 11.2 fold; VCAM-1, 9 fold; P<0.05). This elevated expression was abrogated in the presence of corticosterone at 100 nmol/l. TNF-α significantly increased expression of all inflammatory markers to a much greater degree in K/BxN FLS relative to other mesenchymal cell lines (K/BxN; IL-6, 40.8 fold; CCL-2, 1343.2 fold; VCAM-1, 17.8 fold; ICAM-1, 13.8 fold; P<0.05), with secreted IL-6 mirroring these results (K/BxN; con, 169±29.7 versus TNF-α, 923±378.8 pg/ml/1×10⁵ cells; P<0.05). Dose response experiments confirmed effective concentrations between 10 and 100 nmol/l for corticosterone and 1 and 10 ng/ml for TNF-α, whilst inflammatory gene expression in FLS was shown to be stable between passages four and seven. This study has established a well characterised set of key inflammatory genes for in vitro FLS culture, isolated from K/BxN mice and non-inflamed wild-type controls. Their response to both pro- and anti-inflammatory signalling has been assessed and shown to strongly resemble that which is seen in human FLS culture. Additionally, this study provides guidelines for the effective characterisation, duration and treatment of murine FLS culture.


Valence and diffuse-bound anions of noble-gas complexes with uracil.
Valence-bound (VB) and diffuse-bound (DB) anions of noble-gas (Ar, Kr, and Xe) complexes with uracil have been studied with ab initio methods. MP2 optimizations revealed minima corresponding to anions of both kinds in each case. Coupled-cluster singles and doubles with perturbative triples, CCSD(T), and electron propagator single-point calculations were performed in order to assess vertical and adiabatic electron detachment energies of these complexes. Ab initio electron propagator calculations employed the outer valence Green's function and partial third-order approximations, and the algebraic diagrammatic construction in third order. Basis set effects have been systematically examined. DB anions of all three complexes were adiabatically bound, with calculated adiabatic electron attachment energies below 0.06 eV. Corresponding vertical electron detachment energies were below 0.1 eV. As to VB anions, only the Xe complex had a positive adiabatic electron detachment energy, of 0.01 eV, with a corresponding vertical electron detachment energy of 0.6 eV. These computational findings are consistent with the interpretation of results previously obtained experimentally by Hendricks et al.


Evidence of Energy Transfer from Tryptophan to BSA/HSA Protected Gold Nanoclusters.
This work reports on the chromophores interactions within protein-protected gold nanoclusters. We conducted spectroscopic studies of fluorescence emissions originated from gold nanoclusters and intrinsic tryptophan (Trp) in BSA or HSA proteins. Both, steady state fluorescence and lifetime measurements show a significant Forster resonance energy transfer (FRET) from Trp to the gold nanocluster. Tryptophan lifetimes in the case of protein-protected gold nanoclusters are 2.6ns and 2.3ns for BSA and HSA Au clusters while 5.8ns for native BSA and 5.6 for native HSA. The apparent distances from Trp to gold nanocluster emission center, we estimated as 24.75A0 for BSA and 23.80A0 for HSA. We also studied a potassium iodide (KI) quenching of protein-protected gold nanoclusters and compared with the quenching of BSA and HAS alone. The rates of Trp quenching were smaller in BSA-Au and HSA-Au nanoclusters than in the case of free proteins, which is consistent with shorter lifetime of quenched Trp(s) and lower accessibility for KI. While Trp residues were quenched by KI, the emissions originated from nanoclusters were practically unquenched. In summary, for BSA and HSA Au clusters, we found 55% and 59% energy transfer efficiency respectively from tryoptophan to gold clusters. We believe this interaction can be used to our advantage in terms of developing resonance energy transfer based sensing applications.


Effect of Titrating Positive End-Expiratory Pressure (PEEP) With an Esophageal Pressure-Guided Strategy vs an Empirical High PEEP-Fio2 Strategy on Death and Days Free From Mechanical Ventilation Among Patients With Acute Respiratory Distress Syndrome: A Randomized Clinical Trial.
Adjusting positive end-expiratory pressure (PEEP) to offset pleural pressure might attenuate lung injury and improve patient outcomes in acute respiratory distress syndrome (ARDS). To determine whether PEEP titration guided by esophageal pressure (PES), an estimate of pleural pressure, was more effective than empirical high PEEP-fraction of inspired oxygen (Fio2) in moderate to severe ARDS. Phase 2 randomized clinical trial conducted at 14 hospitals in North America. Two hundred mechanically ventilated patients aged 16 years and older with moderate to severe ARDS (Pao2:Fio2 ≤200 mm Hg) were enrolled between October 31, 2012, and September 14, 2017; long-term follow-up was completed July 30, 2018. Participants were randomized to PES-guided PEEP (n = 102) or empirical high PEEP-Fio2 (n = 98). All participants received low tidal volumes. The primary outcome was a ranked composite score incorporating death and days free from mechanical ventilation among survivors through day 28. Prespecified secondary outcomes included 28-day mortality, days free from mechanical ventilation among survivors, and need for rescue therapy. Two hundred patients were enrolled (mean [SD] age, 56 [16] years; 46% female) and completed 28-day follow-up. The primary composite end point was not significantly different between treatment groups (probability of more favorable outcome with PES-guided PEEP: 49.6% [95% CI, 41.7% to 57.5%]; P = .92). At 28 days, 33 of 102 patients (32.4%) assigned to PES-guided PEEP and 30 of 98 patients (30.6%) assigned to empirical PEEP-Fio2 died (risk difference, 1.7% [95% CI, -11.1% to 14.6%]; P = .88). Days free from mechanical ventilation among survivors was not significantly different (median [interquartile range]: 22 [15-24] vs 21 [16.5-24] days; median difference, 0 [95% CI, -1 to 2] days; P = .85). Patients assigned to PES-guided PEEP were significantly less likely to receive rescue therapy (4/102 [3.9%] vs 12/98 [12.2%]; risk difference, -8.3% [95% CI, -15.8% to -0.8%]; P = .04). None of the 7 other prespecified secondary clinical end points were significantly different. Adverse events included gross barotrauma, which occurred in 6 patients with PES-guided PEEP and 5 patients with empirical PEEP-Fio2. Among patients with moderate to severe ARDS, PES-guided PEEP, compared with empirical high PEEP-Fio2, resulted in no significant difference in death and days free from mechanical ventilation. These findings do not support PES-guided PEEP titration in ARDS. ClinicalTrials.gov Identifier NCT01681225.


Therapeutic HNF4A mRNA attenuates liver fibrosis in a preclinical model.
Therapeutic targeting of injuries that require transient restoration of proteins by mRNA delivery is an attractive approach that, until recently, has remained poorly explored. In this study, we examined the therapeutic utility of mRNA delivery for liver fibrosis and cirrhosis. Specifically, we aimed to demonstrate the therapeutic efficacy of human hepatocyte nuclear factor alpha (HNF4A) mRNA in mouse models of fibrosis and cirrhosis. We investigated restoration of hepatocyte functions by HNF4A mRNA transfection in vitro, and analyzed the attenuation of liver fibrosis and cirrhosis in multiple mouse models, by delivering hepatocyte-targeted biodegradable lipid nanoparticles (LNPs) encapsulating HNF4A mRNA. To identify potential mechanisms of action, we performed microarray-based gene expression profiling, single-cell RNA sequencing, and chromatin immunoprecipitation. We used primary liver cells and human liver buds for additional functional validation. Expression of HNF4A mRNA led to restoration of the metabolic activity of fibrotic primary murine and human hepatocytes in vitro. Repeated in vivo delivery of LNP-encapsulated HNF4A mRNA induced a robust inhibition of fibrogenesis in 4 independent mouse models of hepatotoxin- and cholestasis-induced liver fibrosis. Mechanistically, we discovered that paraoxonase 1 is a direct target of HNF4A and it contributes to HNF4A-mediated attenuation of liver fibrosis via modulation of liver macrophages and hepatic stellate cells. Collectively, our findings provide the first direct preclinical evidence of the applicability of HNF4A mRNA therapeutics for the treatment of fibrosis in the liver. Liver fibrosis and cirrhosis remain unmet medical needs and contribute to high mortality worldwide. Herein, we take advantage of a promising therapeutic approach to treat liver fibrosis and cirrhosis. We demonstrate that restoration of a key gene, HNF4A, via mRNA encapsulated in lipid nanoparticles decreased injury in multiple mouse models of fibrosis and cirrhosis. Our study provides proof-of-concept that mRNA therapy is a promising strategy for reversing liver fibrosis and cirrhosis.


Functional expression and properties of the human skeletal muscle sodium channel.
Full-length deoxyribonucleic acid, complementary (cDNA) constructs encoding the alpha-subunit of the adult human skeletal muscle Na+ channel, hSkM1, were prepared. Functional expression was studied by electrophysiological recordings from cRNA-injected Xenopus oocytes and from transiently transfected tsA201 cells. The Na+ currents of hSkM1 had abnormally slow inactivation kinetics in oocytes, but relatively normal kinetics when expressed in the mammalian cell line. The inactivation kinetics of Na+ currents in oocytes, during a depolarization, were fitted by a weighted sum of two decaying exponentials. The time constant of the fast component was comparable to that of the single component observed in mammalian cells. The block of hSkM1 Na+ currents by the extracellular toxins tetrodotoxin (TTX) and mu-conotoxin (microCTX) was measured. The IC50 values were 25 nM (TTX) and 1.2 microM (microCTX) in oocytes. The potency of TTX is similar to that observed for the rat homolog rSkM1, but the potency of microCTX is 22-fold lower in hSkM1, primarily due to a higher rate of toxin dissociation in hSkM1. Single-channel recordings were obtained from outside-out patches of oocytes expressing hSkM1. The single-channel conductance, 24.9 pS, is similar to that observed for rSkM1 expressed in oocytes.


Dual RNA-Seq transcriptome analysis of chicken macrophage-like cells (HD11) infected in vitro with Eimeria tenella.
The study aimed to monitor parasite and host gene expression during the early stages of Eimeria tenella infection of chicken cells using dual RNA-Seq analysis. For this, we used chicken macrophage-like cell line HD11 cultures infected in vitro with purified E. tenella sporozoites. Cultures were harvested between 2 and 72 h post-infection and mRNA was extracted and sequenced. Dual RNA-Seq analysis showed clear patterns of altered expression for both parasite and host genes during infection. For example, genes in the chicken immune system showed upregulation early (2–4 h), a strong downregulation of genes across the immune system at 24 h and a repetition of early patterns at 72 h, indicating that invasion by a second generation of parasites was occurring. The observed downregulation may be due to immune self-regulation or to immune evasive mechanisms exerted by E. tenella. Results also suggested pathogen recognition receptors involved in E. tenella innate recognition, MRC2, TLR15 and NLRC5 and showed distinct chemokine and cytokine induction patterns. Moreover, the expression of several functional categories of Eimeria genes, such as rhoptry kinase genes and microneme genes, were also examined, showing distinctive differences which were expressed in sporozoites and merozoites.


Rabies: 2015 Update.
Rabies is an almost universally fatal viral disease transmitted to humans primarily by bites and scratches from infected animals, and less commonly through other routes, including transplantation of infected organs, exposure to infected neural tissue, and possibly through airborne and aerosolized routes. This disease is endemic to all continents worldwide except Antarctica, and only a few islands elsewhere can be considered "rabies free." Special Operations Forces medical providers should be aware of this disease. Prevention and recognition of risk are key due to its extreme lethality.


Cyber security threats in the microbial genomics era: implications for public health.
Next generation sequencing (NGS) is becoming the new gold standard in public health microbiology. Like any disruptive technology, its growing popularity inevitably attracts cyber security actors, for whom the health sector is attractive because it combines mission-critical infrastructure and high-value data with cybersecurity vulnerabilities. In this Perspective, we explore cyber security aspects of microbial NGS. We discuss the motivations and objectives for such attack, its feasibility and implications, and highlight policy considerations aimed at threat mitigation. Particular focus is placed on the attack vectors, where the entire process of NGS, from sample to result, could be vulnerable, and a risk assessment based on probability and impact for representative attack vectors is presented. Cyber attacks on microbial NGS could result in loss of confidentiality (leakage of personal or institutional data), integrity (misdetection of pathogens) and availability (denial of sequencing services). NGS platforms are also at risk of being used as propagation vectors, compromising an entire system or network. Owing to the rapid evolution of microbial NGS and its applications, and in light of the dynamics of the cyber security domain, frequent risk assessments should be carried out in order to identify new threats and underpin constantly updated public health policies.


Alternatively spliced androgen receptor variants.
Alternative splicing is an important mechanism for increasing functional diversity from a limited set of genes. Deregulation of this process is common in diverse pathologic conditions. The androgen receptor (AR) is a steroid receptor transcription factor with functions critical for normal male development as well as the growth and survival of normal and cancerous prostate tissue. Studies of AR function in androgen insensitivity syndrome (AIS) and prostate cancer (PCa) have demonstrated loss-of-function AR alterations in AIS and gain-of-function AR alterations in PCa. Over the past two decades, AR gene alterations have been identified in various individuals with AIS, which disrupt normal AR splicing patterns and yield dysfunctional AR protein variants. Recently, altered AR splicing patterns have been identified as a mechanism of PCa progression and resistance to androgen depletion therapy. Several studies have described the synthesis of alternatively spliced transcripts encoding truncated AR isoforms that lack the ligand-binding domain, which is the ultimate target of androgen depletion. Many of these truncated AR isoforms function as constitutively active, ligand-independent transcription factors that can support androgen-independent expression of AR target genes, as well as the androgen-independent growth of PCa cells. In this review, we will summarize the various alternatively spliced AR variants that have been discovered, with a focus on their role and origin in the pathologic conditions of AIS and PCa.