Ligand-free estrogen receptor activity complements IGF1R to induce the proliferation of the MCF-7 breast cancer cells.
Ligand-dependent activation of the estrogen receptor (ER) as well as of the insulin-like growth factor type 1 (IGF1R) induces the proliferation of luminal breast cancer cells. These two pathways cooperate and are interdependent. We addressed the question of the mechanisms of crosstalk between the ER and IGF1R. We evaluated the mitogenic effects of estradiol (E2; agonist ligand of ER) and of insulin (a ligand of IGF1R) in the MCF-7 cells by flow cytometry and by analyzing the cell levels of cell cycle-related proteins (immunoblotting) and mRNA (RT-QPCR). To verify the requirement for the kinase activity of Akt (a downstream target of IGF1R) in the mitogenic action of estradiol, we used shRNA strategy and shRNA-resistant expression vectors. The activation of the ER by E2 is unable to induce the cell cycle progression when the phosphatidyl inositol-3 kinase (PI3K)/Akt signaling is blocked by a chemical inhibitor (LY 294002) or by shRNA targeting Akt1 and Akt2. shRNA-resistant Akt wild-type constructs efficiently complemented the mitogenic signaling activity of E2 whereas constructs with inactivated kinase function did not. In growth factor-starved cells, the residual PI3K/Akt activity is sufficient to complement the mitogenic action of E2. Conversely, when ER function is blocked by the antiestrogen ICI 182780, IGF1R signaling is intact but does not lead to efficient reinitiation of the cell cycle in quiescent, growth factor-starved MCF-7 cells. The basal transcription-promoting activity of ligand-free ER in growth factor-starved cells is sufficient to complement the mitogenic action of the IGF1R-dependent signaling. The basal ER activity in the absence of ligand is sufficient to allow efficient mitogenic action of IGF1R agonists and needs to be blocked to prevent the cell cycle progression.


Effectiveness of Mindfulness-Based Cognitive Therapy for Improving Subjective and Eudaimonic Well-Being in Healthy Individuals: A Randomized Controlled Trial.
Objectives: Better subjective and eudaimonic well-being fosters better health conditions. Several studies have confirmed that mindfulness-based interventions are effective for improving well-being; however, the samples examined in these studies have been limited to specific populations, and the studies only measured certain aspects of well-being rather than the entire construct. Additionally, few studies have examined the effect of mindfulness-based cognitive therapy on well-being. The present study examines the feasibility of mindfulness-based cognitive therapy and its effectiveness for improving subjective and eudaimonic well-being among community residents. Methods: The study design featured an 8-week randomized, waiting-list controlled, parallel-group study. 8 weekly mindfulness classes, followed by 2 monthly classes, were provided for healthy individuals aged 20-65 years who had a Satisfaction with Life Scale score of ≤ 24 indicating average to low cognitive aspect of subjective well-being. This trial was registered with the University Hospital Medical Information Network Clinical Trials Registry (ID: UMIN000031885, URL: https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000036376). Results: The results showed that cognitive aspect of subjective well-being and mindfulness skills were significantly improved at 8 weeks, and this effect was enhanced up to the end of the follow-up period. Positive affective aspect of subjective and eudaimonic well-being were significantly improved at 16 weeks. Conclusions: Eight weeks of mindfulness-based cognitive therapy with a 2-month follow-up period improves cognitive and affective aspects of subjective and eudaimonic well-being in healthy individuals. The order of improvement was cognitive, positive affective, and eudaimonic well-being. To verify these findings, multi-center randomized controlled trials with active control groups and longer follow-up periods are warranted.


Re-assembled casein micelles improve in vitro bioavailability of vitamin D in a Caco-2 cell model.
The pandemic of vitamin D (VD) deficiency, and the global rise in obesity stimulate a need for staple low-fat foods and beverages enriched with VD. In light of consumer demand for a clean label, the use of natural endogenous food ingredients as delivery vehicles is of great interest. To this end, re-assembled casein micelles (rCM) have been shown to help retain VD during processing and shelf life and provide high bioavailability in low-fat milk and non-fat yoghurt. This follow-up study focused on the performance of VD-loaded rCM after drying and reconstitution, considering VD retention during simulated digestion, and the subsequent in vitro bioavailability of the vitamin. rCM conferred great protection to VD3 during simulated digestion with a significant increase in vitamin retention for 1 h under gastric conditions. This observation is believed to arise from the vitamin-casein binding and the system's natural gelation (curd formation) near the casein isoelectric point that seclude the vitamin from environmental stressors and couple its release with digestive proteolysis of the rCM matrix. Vitamin absorption by Caco-2 cells from digested rCM was not significantly different from the absorption of the digested free VD. However, thanks to the highly protective effect of the rCM, against VD gastric degradation, the overall effect of the rCM was a 4-fold higher bioavailability, compared to the free VD.


Temporal regulation of tumor growth in nocturnal mammals: In vivo studies and chemotherapeutical potential.
Tumors of the nervous system including glioblastoma multiforme (GBM) are the most frequent and aggressive form of brain tumors; however, little is known about the impact of the circadian timing system on the formation, growth, and treatment of these tumors. We investigated day/night differences in tumor growth after injection of A530 glioma cells isolated from malignant peripheral nerve sheath tumor (MPNSTs) of NPcis (Trp53+/- ; Nf1+/- ) mice. Synchronized A530 cell cultures expressing typical glial markers were injected at the beginning of the day or night into the sciatic nerve zone of C57BL/6 mice subject to a 12:12 hours light/dark (LD) cycle or after being released to constant darkness (DD). Tumors generated in animals injected early at night in the LD cycle or in DD showed higher growth rates than in animals injected diurnally. No differences were found when animals were injected at the same time with cultures synchronized 12 hours apart. Similar experiments performed with B16 melanoma cells showed higher tumor growth rates in animals injected at the beginning of the night compared to those injected in the daytime. A higher tumor growth rate than that in controls was observed when mice were injected with knocked-down clock gene Bmal1 cells. Finally, when we compared day/night administration of different doses of the proteasome inhibitor Bortezomib (0.5-1.5 mg/kg) in tumor-bearing animals, we found that low-dose chemotherapy displayed higher efficacy when administered at night. Results suggest the existence of a precise temporal control of tumor growth and of drug efficacy in which the host state and susceptibility are critical.


Farnesyltransferase as a target for anticancer drug design.
The currently understood function for Ras in signal transduction is in mediating the transmission of signals from external growth factors to the cell nucleus. Mutated forms of this GTP-binding protein are found in 30% of human cancers with particularly high prevalence in colon and pancreatic carcinomas. These mutations destroy the GTPase activity of Ras and cause the protein to be locked in its active, GTP bound form. As a result, the signaling pathways are activated, leading to uncontrolled tumor growth. Ras function in signaling requires its association with the plasma membrane. This is achieved by posttranslational farnesylation of a cysteine residue present as part of the CA1A2X carboxyl terminal tetrapeptide of all Ras proteins. The enzyme that recognizes and farnesylates the CA1A2X sequence, Ras farnesyltransferase (FTase), has become an important target for the design of inhibitors that might be interesting as antitumor agents. Several approaches have been taken in the search for in vivo active inhibitors of farnesyltransferase. These include the identification of natural products such as the chaetomellic and zaragozic acids that mimic farnesylpyrophosphate, bisubstrate transition state analogs combining elements of the farnesyl and tetrapeptide substrates and peptidomimetics that reproduce features of the carboxyl terminal tetrapeptide CA1A2X sequence. This last group of compounds has been most successful in showing highly potent inhibition of FTase and selective blocking of Ras processing in a range of Ras transformed tumor cell lines at concentrations as low as 10 nM. Certain peptidomimetics will also block tumor growth in various mouse models, with apparently few toxic side effects. These results suggest that farnesyltransferase inhibitors hold considerable promise as anticancer drugs in the clinic.


Nitrogen Limitation Alters Biomass Production but Enhances Steviol Glycoside Concentration in Stevia rebaudiana Bertoni.
The need for medicinal and aromatic plants for industrial uses creates an opportunity for farmers to produce alternative crops. Stevia rebaudiana Bertoni, a perennial shrub originating from Paraguay, is of increasing interest as a source of zero-calorie natural sweeteners: the steviol glycosides (SVglys). The aim of this study was to investigate the relevance of nitrogen (N) supply for leaf yield and for SVgly concentrations in leaves, which are the two major components of S. rebaudiana productivity. In this regard, the relationship between leaf N concentration, CO2 assimilation, leaf production and SVgly accumulation was investigated. The experiments were conducted consecutively in growth-chamber (CC: controlled conditions), in greenhouse (SCC: semi-controlled conditions) and in field conditions (FC) on two genotypes. In CC and SCC, three levels of N fertilization were applied. Plants were grown on four locations in the FC experiment. Both N supply (CC and SCC) and location (FC) had a significant effect on N content in leaves. When light was not limiting (SCC and FC) N content in leaves was positively correlated with CO2 assimilation rate and biomass accumulation. Irrespective of the growth conditions, N content in leaves was negatively correlated with SVgly content. However, increased SVgly content was correlated with a decreased ratio of rebaudioside A over stevioside. The evidence that the increased SVgly accumulation compensates for the negative effect on biomass production suggests that adequate SVgly productivity per plant may be achieved with relatively low fertilization.


Dense cloud cores revealed by CO in the low metallicity dwarf galaxy WLM.
Understanding stellar birth requires observations of the clouds in which they form. These clouds are dense and self-gravitating, and in all existing observations they are molecular, with H2 the dominant species and carbon monoxide (CO) the best available tracer. When the abundances of carbon and oxygen are low compared with that of hydrogen, and the opacity from dust is also low, as in primeval galaxies and local dwarf irregular galaxies, CO forms slowly and is easily destroyed, so it is difficult for it to accumulate inside dense clouds. Here we report interferometric observations of CO clouds in the local group dwarf irregular galaxy Wolf-Lundmark-Melotte (WLM), which has a metallicity that is 13 per cent of the solar value and 50 per cent lower than the previous CO detection threshold. The clouds are tiny compared to the surrounding atomic and H2 envelopes, but they have typical densities and column densities for CO clouds in the Milky Way. The normal CO density explains why star clusters forming in dwarf irregulars have similar densities to star clusters in giant spiral galaxies. The low cloud masses suggest that these clusters will also be low mass, unless some galaxy-scale compression occurs, such as an impact from a cosmic cloud or other galaxy. If the massive metal-poor globular clusters in the halo of the Milky Way formed in dwarf galaxies, as is commonly believed, then they were probably triggered by such an impact.


Chromosome 8p23.1 deletions as a cause of complex congenital heart defects and diaphragmatic hernia.
Recurrent interstitial deletion of a region of 8p23.1 flanked by the low copy repeats 8p-OR-REPD and 8p-OR-REPP is associated with a spectrum of anomalies that can include congenital heart malformations and congenital diaphragmatic hernia (CDH). Haploinsufficiency of GATA4 is thought to play a critical role in the development of these birth defects. We describe two individuals and a monozygotic twin pair discordant for anterior CDH all of whom have complex congenital heart defects caused by this recurrent interstitial deletion as demonstrated by array comparative genomic hybridization. To better define the genotype/phenotype relationships associated with alterations of genes on 8p23.1, we review the spectrum of congenital heart and diaphragmatic defects that have been reported in individuals with isolated GATA4 mutations and interstitial, terminal, and complex chromosomal rearrangements involving the 8p23.1 region. Our findings allow us to clearly define the CDH minimal deleted region on chromosome 8p23.1 and suggest that haploinsufficiency of other genes, in addition to GATA4, may play a role in the severe cardiac and diaphragmatic defects associated with 8p23.1 deletions. These findings also underscore the importance of conducting a careful cytogenetic/molecular analysis of the 8p23.1 region in all prenatal and postnatal cases involving congenital defects of the heart and/or diaphragm.


Knowledge, attitudes and beliefs of primary caretakers towards sickle cell anaemia in children.
The objective of this study was to evaluate knowledge, attitudes and beliefs (KAB) that may influence health seeking behaviour of caretakers of children with sickle cell disease (SCD). A cross-sectional survey was undertaken at Nyanza provincial hospital in Kenya between March and September 1993 to identify socio-demographic and economic factors that may influence health seeking behaviour of primary caretakers of children with SCD. All caretakers accompanying children under the age of 18 years to the Sickle Cell Clinic were eligible. Guardians accompanying children to the clinic were interviewed using pretested questionnaires. An exploratory factor analysis method was used to categorise questionnaire items into domains (knowledge, attitude and belief) and to investigate for association between certain socio-demographic factors and KAB. Seventy five per cent of the 108 respondents interviewed were mothers and 16.7% fathers. Seventy eight percent knew SCD to be hereditary while 55% knew how the disease presents in childhood. Only 42% associated SCD with increased risk of infection. Many felt severe infections are largely preventable and that prevention would reduce their anxiety and illness related costs. In factor analysis, variables loaded almost exclusively on "Attitudes" and "Beliefs" factors. Only family size was found to influence caretaker attitudes (p = 0.0095) and beliefs (p = 0.0034). Education, monthly income, occupation and religion had no significant influence. The majority of caretakers had good knowledge and positive attitudes towards SCD in children. Interventions aimed at management of SCD or prevention of its sequelae would be well accepted. Factor analysis is recommended for statistical analysis of KAB data. The effect of family size on attitudes and behaviour needs further evaluation.


Substrate-selective inhibition of protein kinase PDK1 by small compounds that bind to the PIF-pocket allosteric docking site.
The PIF-pocket of AGC protein kinases participates in the physiologic mechanism of regulation by acting as a docking site for substrates and as a switch for the transduction of the conformational changes needed for activation or inhibition. We describe the effects of compounds that bind to the PIF-pocket of PDK1. In vitro, PS210 is a potent activator of PDK1, and the crystal structure of the PDK1-ATP-PS210 complex shows that PS210 stimulates the closure of the kinase domain. However, in cells, the prodrug of PS210 (PS423) acts as a substrate-selective inhibitor of PDK1, inhibiting the phosphorylation and activation of S6K, which requires docking to the PIF-pocket, but not affecting PKB/Akt. This work describes a tool to study the dynamics of PDK1 activity and a potential approach for drug discovery.


Minocycline and Silver Dual-Loaded Polyphosphoester-Based Nanoparticles for Treatment of Resistant Pseudomonas aeruginosa.
Pseudomonas aeruginosa has been detected in the lungs of ∼50% of patients with cystic fibrosis (CF), including 20% of adult CF patients. The majority of these adult patients harbor multi-drug resistant (MDR) strains, limiting the available treatment options. Silver has long been used as a broad-spectrum antimicrobial agent with a low incidence of resistance. Despite low toxicity, poor availability of silver cations mandates a high dosage to effectively eradicate infections. To address this shortcoming of silver, nanoparticles have been used as delivery devices to improve treatment outcomes. Furthermore, studies have demonstrated that synergistic combinations with careful dose calibrations and efficient delivery systems result in superior antimicrobial activity while avoiding potential side effects of both therapeutics. Here 4-epi-minocycline, a metabolite of minocycline, was identified as an active antimicrobial against P. aeruginosa using a high-throughput screen. The antimicrobial activities of 4-epi-minocycline, minocycline, and silver acetate against clinical isolates of P. aeruginosa obtained from CF patients were evaluated in vitro. Next, the synergistic activity of the silver/minocycline combination against P. aeruginosa isolates was investigated using checkerboard assays and identified with end-point colony forming unit determination assays. Finally, nanoparticles coloaded with minocycline and silver were evaluated in vitro for antimicrobial activity. The results demonstrated that both silver and minocycline are potent antimicrobials alone and that the combination allows a reduced dosage of both therapeutics to achieve the same antimicrobial effect. Furthermore, the proposed synergistic silver/minocycline combination can be coloaded into nanoparticles as a next-generation antibiotic to combat the threats presented by MDR pathogens.


[Surgical anatomy of the anterior mediastinum].
The mediastinum is located from the thoracic inlet to the diaphragm between the left and right pleural cavities and contains vital structures of the circulatory, respiratory, digestive, and nervous system. Over the years, since there are no fascial or anatomic planes, anatomists and radiologists have suggested various schemes for subdividing the mediastinum and several anatomical and radiological classifications of the mediastinum are reported in the literature. The most popular of these scheme divides medistinum, for purposes of description, into two parts: an upper portion, above the upper level of the pericardium, which is named the superior mediastinum; and a lower portion, below the upper level of the pericardium. For clinical purposes, the mediastinum may be subdivided into three major areas, i.e. anterior, middle, and posterior compartments. The anterior mediastinum is defined as the region posterior to the sternum and anterior to the heart and brachiocephalic vessels. It extends from the thoracic inlet to the diaphragm and contains the thymus gland, fat, and lymph nodes. This article will review surgical anatomy of the anterior mediastinum and will focus on the surgical approch to anterior mediastinum and thymic diseases.


Are lumbar spine reoperation rates falling with greater use of fusion surgery and new surgical technology?
A retrospective analysis of population-based hospital discharge registry from all nonfederal acute care hospitals in Washington State. We examined the cumulative incidence of second lumbar spine operation following an initial lumbar operation for degenerative conditions. We aimed to determine if the cumulative incidence of a second lumbar spine operation decreased in the 1990s following an increase in the rate of fusion surgery and the introduction of several newer fusion technologies. Repeat lumbar spine operations are generally undesirable, implying persistent symptoms, progression of degenerative changes, or treatment complications. Improved technology is expected to improve alignment, healing, and instability, and to reduce repeat operations. Among the patients who had an inpatient lumbar decompression or lumbar fusion surgery for degenerative spine disorders in 1990 to 1993 (n = 24,882) or in 1997 to 2000 (n = 25,209), we examined rates of subsequent lumbar spine surgery during a 4-year follow-up. We performed a Cox proportional hazards regression to compare the probability of a reoperation between the 2 cohorts, adjusting for age, sex, primary diagnosis, type of insurance, and comorbidity. Among patients who underwent surgery for lumbar degenerative disease, more than twice as many had a fusion procedure in the 1997 to 2000 cohort (19.1%) compared with the 1990 to 1993 cohort (9.4%). However, the 4-year cumulative incidence of reoperation was higher in the 1997 to 2000 cohort compared with the 1990 to 1993 cohort (14.0% vs. 12.4%; hazard ratio, 1.16; 95% confidence interval, 1.11-1.22, P < 0.001). Among fusion patients, those in the 1997 to 2000 cohort were approximately 40% more likely to undergo a reoperation within the first year when compared with fusion patients in the 1990 to 1993 cohort. There was no difference in reoperation probability beyond 1 year. A higher proportion of fusion procedures and the introduction of new spinal implants between 1993 and 1997 did not reduce reoperation rates. Patients who had lumbar surgery for degenerative disease in the late 1990s were more likely to undergo a repeat operation within 4 years than patients who had surgery in the early 1990s.


RE-AIM in the Real World: Use of the RE-AIM Framework for Program Planning and Evaluation in Clinical and Community Settings.
Background: The RE-AIM framework has been widely used in health research but it is unclear the extent to which this framework is also used for planning and evaluating health-related programs in clinical and community settings. Our objective was to evaluate how RE-AIM is used in the "real-world" and identify opportunities for improving use outside of research contexts. Methods: We used purposive and snowball sampling to identify clinical and community health programs that used RE-AIM for planning and/or evaluation. Recruitment methods included surveys with email follow-up to funders, implementers, and RE-AIM working group members. We identified 17 programs and conducted structured in-depth interviews with key informants (n = 18). Across RE-AIM dimensions, respondents described motivations, uses, and measures; rated understandability and usefulness; discussed benefits and challenges, strategies to overcome challenges, and resources used. We used descriptive statistics for quantitative ratings, and content analysis for qualitative data. Results: Program content areas included chronic disease management and prevention, healthy aging, mental health, or multiple, often behavioral health-related topics. During planning, most programs considered reach (n = 9), adoption (n = 11), and implementation (n = 12) while effectiveness (n = 7) and maintenance (n = 6) were considered less frequently. In contrast, most programs evaluated all RE-AIM dimensions, ranging from 13 programs assessing maintenance to 15 programs assessing implementation and effectiveness. On five-point scales, all RE-AIM dimensions were rated as easy to understand (Overall M = 4.7 ± 0.5), but obtaining data was rated as somewhat challenging (Overall M = 3.4 ± 0.9). Implementation was the most frequently used dimension to inform program design (M = 4.7 ± 0.6) relative to the other dimensions (3.0-3.9). All dimensions were considered similarly important for decision-making (average M = 4.1 ± 1.4), with the exception of maintenance (M = 3.4 ± 1.7). Qualitative corresponded to the quantitative findings in that RE-AIM was reported to be a practical, easy to understand, and well-established implementation science framework. Challenges included understanding differences among RE-AIM dimensions and data acquisition. Valuable resources included the RE-AIM website and collaborating with an expert. Discussion: RE-AIM is an efficient framework for planning and evaluation of clinical and community-based projects. It provides structure to systematically evaluate health program impact. Programs found planning for and assessing maintenance difficult, providing opportunities for further refinement.


Injury Response of Resected Human Brain Tissue In Vitro.
Brain injury affects a significant number of people each year. Organotypic cultures from resected normal neocortical tissue provide unique opportunities to study the cellular and neuropathological consequences of severe injury of adult human brain tissue in vitro. The in vitro injuries caused by resection (interruption of the circulation) and aggravated by the preparation of slices (severed neuronal and glial processes and blood vessels) reflect the reaction of human brain tissue to severe injury. We investigated this process using immunocytochemical markers, reverse transcriptase quantitative polymerase chain reaction and Western blot analysis. Essential features were rapid shrinkage of neurons, loss of neuronal marker expression and proliferation of reactive cells that expressed Nestin and Vimentin. Also, microglia generally responded strongly, whereas the response of glial fibrillary acidic protein-positive astrocytes appeared to be more variable. Importantly, some reactive cells also expressed both microglia and astrocytic markers, thus confounding their origin. Comparison with post-mortem human brain tissue obtained at rapid autopsies suggested that the reactive process is not a consequence of epilepsy.


Gravity-driven microfluidic assay for digital enumeration of bacteria and for antibiotic susceptibility testing.
The alarming dynamics of antibiotic-resistant infections calls for the development of rapid and point-of-care (POC) antibiotic susceptibility testing (AST) methods. Here, we demonstrated the first completely stand-alone microfluidic system that allowed the execution of digital enumeration of bacteria and digital antibiograms without any specialized microfluidic instrumentation. A four-chamber gravity-driven step emulsification device generated ∼2000 monodisperse 2 nanoliter droplets with a coefficient of variation of 8.9% of volumes for 95% of droplets within less than 10 minutes. The manual workload required for droplet generation was limited to the sample preparation, the deposition into the sample inlet of the chip and subsequent orientation of the chip vertically without an additional pumping system. The use of shallow chambers imposing a 2D droplet arrangement provided superior stability of the droplets against coalescence and minimized the leakage of the reporter viability dye between adjacent droplets during long-term culture. By using resazurin as an indicator of the growth of bacteria, we were also able to reduce the assay time to ∼5 hours compared to 20 hours using the standard culture-based test.


Metrics of optical quality derived from wave aberrations predict visual performance.
Wavefront-guided refractive surgery and custom optical corrections have reduced the residual root mean squared (RMS) wavefront error in the eye to relatively low levels (typically on the order of 0.25 microm or less over a 6-mm pupil, a dioptric equivalent of 0.19 D). It has been shown that experimental variation of the distribution of 0.25 microm of wavefront error across the pupil can cause variation in visual acuity of two lines on a standard logMAR acuity chart. This result demonstrates the need for single-value metrics other than RMS wavefront error to quantify the effects of low levels of aberration on acuity. In this work, we present the correlation of 31 single-value metrics of optical quality to high-contrast visual acuity for 34 conditions where the RMS wavefront error was equal to 0.25 microm over a 6-mm pupil. The best metric, called the visual Strehl ratio, accounts for 81% of the variance in high-contrast logMAR acuity.


The α7 nicotinic acetylcholine receptor positive allosteric modulator attenuates lipopolysaccharide-induced activation of hippocampal IκB and CD11b gene expression in mice.
We have reported that 3a,4,5,9b-tetrahydro-4-(1-naphthalenyl)-3H-cyclopentan[c]quinoline-8-sulfonamide (TQS), α7 nicotinic acetylcholine receptor (nAChR) positive allosteric modulator (PAM) reduces lipopolysaccharide (LPS)-induced hyperalgesia and allodynia in mice. The objective of the present study was to determine the effects of TQS on LPS-induced activation of hippocampal inhibitor of κB (IκB) and cluster of differentiation 11b (CD11b) gene expression involving hyperalgesia and allodynia in mice. We also examined the effects of TQS on microglial phenotype following LPS administration. Pretreatment of TQS (4 mg/kg) reduced the expressions of IκB and CD11b mRNA. Pretreatment of methyllycaconitine (3 mg/kg), an α7 nAChR antagonist, reversed TQS-induced decrease in IκB and CD11b mRNA expressions in the hippocampus indicating the involvement of α7 nAChR. In addition, TQS (4 mg/kg) reversed the LPS-induced microglial morphological changes. These results suggest that TQS reduces LPS-induced IκB and CD11b gene expression and microglial activation associated with hyperalgesia and allodynia by targeting microglial α7 nAChR in the hippocampus.


The Efficacy of an In-Service Education Program Designed to Enhance the Effectiveness of Physical Restraints.
Physical restraints are used to enhance the safety of patients and to avoid injury. However, physical restraints may cause injuries if improperly used or if they are used in the absence of continuous monitoring. Nursing staff who use physical restraints often lack sufficient related knowledge, which may increase the risk to patient safety. This study investigates the impact of an in-service education program for nursing staff that is designed to improve physical-restraint-related knowledge, attitudes, behaviors, and techniques. A pretest-posttest design and a quasi-experimental method were employed to evaluate the effectiveness of the in-service education program. One hundred thirty-six nursing staff from four adult intensive care units (ICUs), including two medical ICUs and two surgical ICUs, in a medical center in central Taiwan were enrolled as participants. The experimental group (EG) and the control group (CG) were composed of patients from one randomly assigned medical ICU and one randomly assigned surgical ICU each. The pretest data on physical-restraint-related knowledge, attitudes, behaviors, and techniques were collected before the in-service education program. The EG received 2 hours of classroom education on guidelines and techniques related to physical restraints. The posttest data for the two groups were collected a month after implementation of the in-service education program. General Estimation Equation was used to measure and analyze the data repeatedly. The posttest scores of the EG for knowledge and technique were significantly higher than the pretest scores (p < .0001). However, the posttest scores of the EG for attitudes and behaviors did not significantly differ from the pretest scores. In-service education for physical restraints enhances relevant knowledge and techniques but does not significantly affect attitudes or behaviors. Correct implementation of physical restraints not only promotes the quality of nursing care for patients in the ICU but also reduces the risk of physical-restraint-related complications. This study highlights the importance of changing the thoughts and concepts related to the use of physical restraints within the overall caring strategy of hospitals.


Screening of patients with tuberculosis for diabetes mellitus in China.
There is a high burden of both diabetes (DM) and tuberculosis (TB) in China, and this study aimed to assess feasibility and results of screening patients with TB for DM within the routine healthcare setting of six health facilities. Agreement on how to screen, monitor and record was reached in May 2011 at a stakeholders' meeting, and training was carried out for staff in the six facilities in July 2011. Implementation started in September 2011, and we report on 7 months of activities up to 31 March 2012. There were 8886 registered patients with TB. They were first asked whether they had DM. If the answer was no, they were screened with a random blood glucose (RBG) followed by fasting blood glucose (FBG) in those with RBG ≥ 6.1 mm (one facility) or with an initial FBG (five facilities). Those with FBG ≥ 7.0 mm were referred to DM clinics for diagnostic confirmation with a second FBG. Altogether, 1090 (12.4%) patients with DM were identified, of whom 863 (9.7%) had a known diagnosis of DM. Of 8023 patients who needed screening for DM, 7947 (99%) were screened. This resulted in a new diagnosis of DM in 227 patients (2.9% of screened patients), and of these, 226 were enrolled to DM care. In addition, 575 (7.8%) persons had impaired fasting glucose (FBG 6.1 to <7.0 mm). Prevalence of DM was significantly higher in patients in health facilities serving urban populations (14.0%) than rural populations (10.6%) and higher in hospital patients (13.5%) than those attending TB clinics (8.5%). This pilot project shows that it is feasible to screen patients with TB for DM in the routine setting, resulting in a high yield of patients with known and newly diagnosed disease. Free blood tests for glucose measurement and integration of TB and DM services may improve the diagnosis and management of dually affected patients.


Etiology and clinical outcomes of acute respiratory virus infection in hospitalized adults.
Etiologies and clinical profiles of acute respiratory viral infections need to be clarified to improve preventive and therapeutic strategies. A retrospective observational study at a single, university-affiliated center was performed to evaluate the respiratory viral infection etiologies in children compared to that in adults and to document the clinical features of common viral infections for adults from July 2009 to April 2012. The common viruses detected from children (2,800 total patients) were human rhinovirus (hRV) (31.8%), adenovirus (AdV) (19.2%), respiratory syncytial virus (RSV) A (17.4%), RSV B (11.7%), and human metapneumovirus (hMPV) (9.8%). In comparison, influenza virus A (IFA) had the highest isolation rate (28.5%), followed by hRV (15.5%), influenza virus B (IFB) (15.0%), and hMPV (14.0%), in adults (763 total patients). Multiple viruses were detected in single specimens from 22.4% of children and 2.0% of adults. IFA/IFB, RSV A/B, and hMPV exhibited strong seasonal detection and similar circulating patterns in children and adults. Adult patients showed different clinical manifestations according to causative viruses; nasal congestion and rhinorrhea were more common in hRV and human coronavirus (hCoV) infection. Patients with RSV B, hRV, or AdV tended to be younger, and those infected with RSV A and hMPV were likely to be older. Those with RSV A infection tended to stay longer in hospital, enter the intensive care unit more frequently, and have a fatal outcome more often. The bacterial co-detection rate was 26.5%, and those cases were more likely to have lower respiratory tract involvement (P = 0.001), longer hospital stay (P = 0.001), and higher mortality (P = 0.001). The etiologic virus of an acute respiratory infection can be cautiously inferred based on a patient's age and clinical features and concurrent epidemic data. Large-scale prospective surveillance studies are required to provide more accurate information about respiratory viral infection etiology, which could favorably influence clinical outcomes.


Evidence for selective evolution in codon usage in conserved amino acid segments of human alphaherpesvirus proteins.
The genomes of human viruses herpes simplex 1 (HSV1) and varicella zoster (VZV), although similar in biology, largely concordant in gene order, and identical in many amino acid segments, differ widely in their genomic G + C (abbreviated S) content, which is high in HSV1 (68%) and low in VZV (46%). This paper analyzes several striking codon usage contrasts. The S difference in coding regions is dramatically large in codon site 3, S3, about 42%. The large difference in S3 is maintained at the same level in a subset of closely similar genes and even in corresponding identical amino acid blocks. A similar difference in S levels in silent site 1 (S1) is found in leucine and arginine. The difference in S3 levels occurs in every gene and in every multicodon amino acid form. The S difference also exists in amino acid usage, with HSV1 using significantly more codon types SSN, while VZV uses more codon types WWN (where W stands for A or T). The nonoverlapping and narrow histograms of S3 gene frequencies in both viruses suggest that the difference has arisen and been maintained by a process of selective rather than nonselective effects. This is in sharp contrast to the relatively large variance seen for highly similar genes in the human versus yeast analysis. Interpretations and hypotheses to explain the HSV1 vs VZV codon usage disparity relate to virus-host interactions, to the role of viral genes in DNA metabolism, to availability of molecular resources (molecular Gause exclusion principle), and to differences in genomic structure.


A positive feedback loop stabilizes the guanine-nucleotide exchange factor Cdc24 at sites of polarization.
In Saccharomyces cerevisiae, activation of Cdc42 by its guanine-nucleotide exchange factor Cdc24 triggers polarization of the actin cytoskeleton at bud emergence and in response to mating pheromones. The adaptor protein Bem1 localizes to sites of polarized growth where it interacts with Cdc42, Cdc24 and the PAK-like kinase Cla4. We have isolated Bem1 mutants (Bem1-m), which are specifically defective for binding to Cdc24. The mutations map within the conserved PB1 domain, which is necessary and sufficient to interact with the octicos peptide repeat (OPR) motif of Cdc24. Although Bem1-m mutant proteins localize normally, bem1-m cells are unable to maintain Cdc24 at sites of polarized growth. As a consequence, they are defective for apical bud growth and the formation of mating projections. Localization of Bem1 to the incipient bud site requires activated Cdc42, and conversely, expression of Cdc42-GTP is sufficient to accumulate Bem1 at the plasma membrane. Thus, our results suggest that Bem1 functions in a positive feedback loop: local activation of Cdc24 produces Cdc42-GTP, which recruits Bem1. In turn, Bem1 stabilizes Cdc24 at the site of polarization, leading to apical growth.


Alterations in the gut bacterial microbiome in fungal Keratitis patients.
Dysbiosis in the gut microbiome has been implicated in several diseases including auto-immune diseases, inflammatory diseases, cancers and mental disorders. Keratitis is an inflammatory disease of the eye significantly contributing to corneal blindness in the developing world. It would be worthwhile to investigate the possibility of dysbiosis in the gut microbiome being associated with Keratitis. Here, we have analyzed fungal and bacterial populations in stool samples through high-throughput sequencing of the ITS2 region for fungi and V3-V4 region of 16S rRNA gene for bacteria in healthy controls (HC, n = 31) and patients with fungal keratitis (FK, n = 32). Candida albicans (2 OTUs), Aspergillus (1 OTU) and 3 other denovo-OTUs were enriched in FK samples and an unclassified denovo-OTU was enriched in HC samples. However, the overall abundances of these 'discriminatory' OTUs were very low (< 0.001%) and not indicative of significant dysbiosis in the fungal community inhabiting the gut of FK patients. In contrast, the gut bacterial richness and diversity in FK patients was significantly decreased when compared to HC. 52 OTUs were significantly enriched in HC samples whereas only 5 OTUs in FK. The OTUs prominently enriched in HC were identified as Faecalibacterium prausnitzii, Bifidobacterium adolescentis, Lachnospira, Mitsuokella multacida, Bacteroides plebeius, Megasphaera and Lachnospiraceae. In FK samples, 5 OTUs affiliated to Bacteroides fragilis, Dorea, Treponema, Fusobacteriaceae, and Acidimicrobiales were significantly higher in abundance. The functional implications are that Faecalibacterium prausnitzii, an anti-inflammatory bacterium and Megasphaera, Mitsuokella multacida and Lachnospira are butyrate producers, which were enriched in HC patients, whereas Treponema and Bacteroides fragilis, which are pathogenic were abundant in FK patients, playing a potential pro-inflammatory role. Heatmap, PCoA plots and functional profiles further confirm the distinct patterns of gut bacterial composition in FK and HC samples. Our study demonstrates dysbiosis in the gut bacterial microbiomes of FK patients compared to HC. Further, based on inferred functions, it appears that dysbiosis in the gut of FK subjects is strongly associated with the disease phenotype with decrease in abundance of beneficial bacteria and increase in abundance of pro-inflammatory and pathogenic bacteria.


[Effects of sperminated pullulans on the pulmonary absorption of insulin].
Sperminated pullulans (SP) having different molecular weights (MWs) were prepared, and the enhancing effect on the pulmonary absorption of insulin in rats was examined. SP acted as enhancers of insulin absorption when a 0.1% solution was applied with insulin simultaneously and their enhancing effects depended on the MW of the SP; the same solutions exhibited low toxicity in the in vivo LDH leaching test. In the in vitro experiments using Calu-3 cells, tight junction-opening effects and a toxic effect of SP in the MTT assay were observed at lower concentrations compared with the in vivo experiments. A mucus layer might interfere with the interaction between SP and the cell surface and might suppress both these effects and toxicity. SP having a high MW will be useful for preparing safe and efficient formulations of peptide and protein drugs. The change in the localization of the tight junction proteins may be related to the permeation-enhancing mechanism of SP.


Modal amplification in active waveguides with hyperbolic dispersion at telecommunication frequencies.
We present a method for studying amplification of electromagnetic modes in active, circularly symmetric waveguides with hyperbolic dispersion. Using this method, we obtain a closed-form expression for the modal threshold condition. We find that modal amplification is possible in a region of the radius-wavelength phase-space with small enough radius so that propagation of the mode is permitted while modal energy and phase counter-propagate. At telecommunication frequencies, such a situation is achievable only when the absolute value of the real metal permittivity exceeds that of the active dielectric. We validate our theoretical conclusions with numerical simulations that explain the threshold condition in terms of an energy balance between the longitudinal and radial components of the electric field.


The BIOSAFEPAPER project for in vitro toxicity assessments: preparation, detailed chemical characterisation and testing of extracts from paper and board samples.
Nineteen food contact papers and boards and one non-food contact board were extracted following test protocols developed within European Union funded project BIOSAFEPAPER. The extraction media were either hot or cold water, 95% ethanol or Tenax, according to the end use of the sample. The extractable dry matter content of the samples varied from 1200 to 11,800 mg/kg (0.8-35.5 mg/dm2). According to GC-MS the main substances extracted into water were pulp-derived natural products such as fatty acids, resin acids, natural wood sterols and alkanols. Substances extracted into ethanol particularly, were diisopropylnaphthalenes, alkanes and phthalic acid esters. The non-food contact board showed the greatest number and highest concentrations of GC-MS detectable compounds. The extracts were subjected to a battery of in vitro toxicity tests measuring both acute and sublethal cytotoxicity and genotoxic effects. None of the water or Tenax extracts was positive in cytotoxicity or genotoxicity assays. The ethanol extract of the non-food contact board gave a positive response in the genotoxicity assays, and all four ethanol extracts gave positive response(s) in the cytotoxicity assays to some extent. These responses could not be pinpointed to any specific compound, although there appeared a correlation between the total amount of extractables and toxicity.


National health workforce in discrete Indigenous communities.
To identify areas of relative need and inform future planning of health workforce and health services in discrete Indigenous communities. Descriptive analysis of relevant variables from the 1999 Community Housing and Infrastructure Needs Survey (CHINS), including all discrete Indigenous communities in Australia. Almost 90% of the Indigenous population of the Northern Territory live in discrete communities. The corresponding figure for Queensland, South Australia and Western Australia is around 25%, for New South Wales 8% and Victoria 1%. Just over 4000 people (5% of the population surveyed) live 100 kilometres or more from the nearest community health centre and almost 60,000 (54%) live 100 kilometres or more from the nearest hospital. Approximately 4000 Indigenous people (6% of population surveyed) have little or no access to a registered nurse or a doctor in their community. Access to Indigenous health workers is also limited, with more than 26,000 people (40%) having almost no access to a male Indigenous health worker and about 10,400 (16%) having almost no access to a female Indigenous health worker. More than 13,000 people (20%) have no access to a dentist and many thousands (30-50%) have no access to allied health or mental health care workers. An obstetrician or ENT/respiratory physician never visited the communities of almost 40,000 people (55% and 59%, respectively) and about 24,000 people (36%) have no access to an ophthalmologist. CHINS data provide a unique source of information to monitor the status of health services and the workforce in discrete Indigenous communities.


Effects of external feedback about body tilt: Influence on the Subjective Proprioceptive Horizon.
The present study investigated a cognitive aspect upon spatial perception, namely the impact of a true or false verbal feedback (FB) about the magnitude of body tilt on Subjective Proprioceptive Horizon (SPH) estimates. Subjects were asked to set their extended arm normal to gravity for different pitch body tilts up to 9 degrees . True FB were provided at all body tilt angles, whereas false FB were provided only at 6 degrees backward and 6 degrees forward body tilts for half of the trials. Our data confirmed previous results about the egocentric influence of body tilt itself upon SPH: estimates were linearly lowered with forward tilts and elevated with backward tilts. In addition, results showed a significant effect of the nature of the external FB provided to the subjects. When subjects received a false FB inducing a 3 degrees forward bias relative to physical body tilt, they set their SPH consequently higher than when they received a false FB inducing a 3 degrees backward bias. These findings clearly indicated that false cognitive information about body tilt might significantly modify the judgement of a geocentric direction of space, such as the SPH. This may have deleterious repercussions in aeronautics when pilots have to localize external objects relative to earth-based directions in darkened environments.


Homologous and cross-reactive precipitins in anti-pneumococcal sera raised in mules.
Serial bleedings were obtained from two mules during prolonged immunization, one with type XXV the other with type VIII pneumococcal vaccine. IgGa, IgGb, IgGc, IgB, IgG(T) and IgM present among purified Pn anti-XXV and Pn anti-VIII immunoglobulin isolated from various bleedings were identified by use of rabbit anti-equine heavy chain specific reagents. Radioimmunodiffusion with 14C-labelled type XXV pneumococcal capsular polysaccharide and horse and donkey reagents with species specificity directed against donkey or horse IgGa respectively, demonstrated both parental horse and donkey IgGa heavy chain isotypes among the anti-PnXXV antibodies of the interspecies hybrid. Qualtitative and quantitative examination of the cross-precipitation of mule anti-PnXXV sera with the capsular polysaccharides of pneumococcal types IV, X and XA, with birch sap, ketha gum, and with polysaccharides of E. coli, Klebsiella and Rhizobium was carried out and compared with data obtained with anti-PnXXV raised in a horse. Analysis of supernatants from the cross-reactions showed that distinct subfractions had reacted. indicating a marked heterogeneity of the antibodies.


Drug-drug interaction between cannabidiol and phenobarbital in healthy dogs.
To assess drug-drug interactions between cannabidiol (CBD) and phenobarbital (PB) when simultaneously administered to healthy dogs. 9 healthy, purpose bred Beagles. A 3-phase prospective, randomized pharmacokinetic (PK) interaction study of CBD and PB was performed as follows: phase 1, CBD PK determination and evaluation of CBD tolerability by 3 single-dose CBD (5 mg/kg, 10 mg/kg, and 20 mg/kg) protocols followed by 2-week CBD dosing; phase 2, a single-dose, 3-way, crossover PK study of CBD (10 mg/kg), PB (4 mg/kg), or CBD (10 mg/kg) administration plus PB (4 mg/kg); and phase 3, evaluation of chronic PB (4 mg/kg, q 30 d) administration followed by single-dose CBD (10 mg/kg) PK study. Although there were variations in CBD PK variables in dogs receiving CBD alone or in conjunction with PB, significance differences in CBD PK variables were not found. No significant difference was observed in PB PK variables of dogs receiving PB alone or with CBD. During chronic CBD administration, mild gastrointestinal signs were observed in 5 dogs. At daily CBD doses of 10 to 20 mg/kg/d, hypoxia was observed in 5 dogs and increased serum alkaline phosphatase (ALP) activities (range, 301 to 978 U/L) was observed in 4 dogs. A significant increase in ALP activity was observed with chronic administration of CBD during phase 1 between day 0 and day 14. No significant PK interactions were found between CBD and PB. Dose escalation of CBD or adjustment of PB in dogs is not recommended on the basis of findings of this study.


Asthma risk factor assessment: what are the needs of inner-city families?
A complex array of risk factors contributes to sustained high levels of asthma morbidity in inner-city children. To describe risk factors for asthma morbidity in a national sample of inner-city children with persistent asthma. This study examined baseline questionnaire results from 1,772 children ages 5 to 11 years old with moderate to severe persistent asthma who enrolled in the Centers for Disease Control and Prevention-funded Inner-City Asthma Intervention between April 2001 and March 2004. Risk for asthma morbidity was assessed in 9 domains using the Child Asthma Risk Assessment Tool. The domains included environmental exposures, parental stress, medication adherence, pessimistic asthma beliefs, smoke exposure, aeroallergen exposure, child psychological well-being, responsibility for medication administration, and medical care. A total of 51% of families demonstrated high risk of asthma morbidity in 3 or more domains. High risk of asthma morbidity was suggested based on household environmental exposures (47.7%), high parental stress (38.5%), poor medication adherence (38.3%), pessimistic asthma beliefs (31.8%), environmental tobacco smoke (24.4%), sensitization to aeroallergens in the home (24.8%), child behavioral or emotional concerns (22.9%), child assigned responsibility for medication administration (21.2%), and poor medical care (20.7%). Allergy testing was completed for 40% of the participating children. Of these children, 61% were exposed to aeroallergens in their home to which they were sensitized. In this national sample of inner-city children, multiple risk factors for asthma morbidity were identified. Asthma programs that provide multilevel support and intervention are needed to reduce the burden of asthma on inner-city families.


Integrating Sphere Microscopy for Direct Absorption Measurements of Single Nanostructures.
Nanoscale materials are promising for optoelectronic devices because their physical dimensions are on the order of the wavelength of light. This leads to a variety of complex optical phenomena that, for instance, enhance absorption and emission. However, quantifying the performance of these nanoscale devices frequently requires measuring absolute absorption at the nanoscale, and remarkably, there is no general method capable of doing so directly. Here, we present such a method based on an integrating sphere but modified to achieve submicron spatial resolution. We explore the limits of this technique by using it to measure spatial and spectral absorptance profiles on a wide variety of nanoscale systems, including different combinations of weakly and strongly absorbing and scattering nanomaterials (Si and GaAs nanowires, Au nanoparticles). This measurement technique provides quantitative information about local optical properties that are crucial for improving any optoelectronic device with nanoscale dimensions or nanoscale surface texturing.


Modelling the distributions of Culicoides bluetongue virus vectors in Sicily in relation to satellite-derived climate variables.
Surveillance data from 268 sites in Sicily are used to develop climatic models for prediction of the distribution of the main European bluetongue virus (BTV) vector Culicoides imicola Kieffer (Diptera: Ceratopogonidae) and of potential novel vectors, Culicoides pulicaris Linnaeus, Culicoides obsoletus group Meigen and Culicoides newsteadi Austen. The models containing the 'best' climatic predictors of distribution for each species, were selected from combinations of 40 temporally Fourier-processed remotely sensed variables and altitude at a 1 km spatial resolution using discriminant analysis. Kappa values of around 0.6 for all species models indicated substantial levels of agreement between model predictions and observed data. Whilst the distributions of C. obsoletus group and C. newsteadi were predicted by temperature variables, those of C. pulicaris and C. imicola were determined mainly by normalized difference vegetation index (NDVI), a variable correlated with soil moisture and vegetation biomass and productivity. These models were used to predict species presence in unsampled pixels across Italy and for C. imicola across Europe and North Africa. The predicted continuous presence of C. pulicaris along the appenine mountains, from north to south Italy, suggests BTV transmission may be possible in a large proportion of this region and that seasonal transhumance (seasonal movement of livestock between upland and lowland pastures) even in C. imicola-free areas should not generally be considered safe. The predicted distribution of C. imicola distribution shows substantial agreement with observed surveillance data from Greece and Iberia (including the Balearics) and parts of mainland Italy (Lazio, Tuscany and areas of the Ionian coast) but is generally much more restricted than the observed distribution (in Sardinia, Corsica and Morocco). The low number of presence sites for C. imicola in Sicily meant that only a restricted range of potential C. imicola habitats were included in the training set and that predictions could only be made within this range. Future modelling exercises will use abundance data collected according to a standardized protocol across the Mediterranean and, for Sicily in particular, should include non-climatic environmental variables that may influence breeding site suitability such as soil type.


Proneness to decreased negative emotions in major depressive disorder when blaming others rather than oneself.
One widespread view holds that vulnerability to major depressive disorder (MDD) is linked to overall increases in negative emotionality. In contrast, cognitive attribution theories emphasize the importance of blaming oneself rather than others for negative events. Thus far, the contrasting predictions of these models have not been directly compared. Following the attributional perspective, we tested the hypothesis that people with remitted MDD show no overall bias towards negative emotions, but a selective bias towards self-blaming emotions relative to those emotions associated with blaming others. We compared a remitted MDD and a control group on a novel experimental test that allowed us to directly compare proneness to specific emotions associated with different types of self-blame (guilt, shame, self-contempt/disgust) and blame of others (other-indignation/anger, other-contempt/disgust) whilst controlling for negative valence and medication status, and excluding comorbidity. In agreement with our hypothesis, individuals with remitted MDD exhibited an increased self-contempt bias (difference between contempt/disgust towards self and others) but no increased proneness to any other negative emotion or overall increases in perceived negative valence of stimuli. Moreover, the remitted MDD group exhibited reduced contempt/disgust towards others. Our results corroborate the prediction that vulnerability to MDD is associated with an imbalance of specific self- and other-blaming emotions rather than a general increase in negative emotions. Based on the composition of our sample, we speculate that self-contempt bias may be particularly characteristic of melancholic MDD subtypes and could be useful for stratification of depression in the future.


Investigating Therapeutic Potential of Trigonella foenum-graecum L. as Our Defense Mechanism against Several Human Diseases.
Current lifestyle, stress, and pollution have dramatically enhanced the progression of several diseases in human. Globally, scientists are looking for therapeutic agents that can either cure or delay the onset of diseases. Medicinal plants from time immemorial have been used frequently in therapeutics. Of many such plants, fenugreek is one of the oldest herbs which have been identified as an important medicinal plant by the researchers around the world. It is potentially beneficial in a number of diseases such as diabetes, hypercholesterolemia, and inflammation and probably in several kinds of cancers. It has industrial applications such as synthesis of steroidal hormones. Its medicinal properties and their role in clinical domain can be attributed to its chemical constituents. The 3 major chemical constituents which have been identified as responsible for principle health effects are galactomannan, 4-OH isoleucine, and steroidal saponin. Numerous experiments have been carried out in vivo and in vitro for beneficial effects of both the crude chemical and of its active constituent. Due to its role in health care, the functional food industry has referred to it as a potential nutraceutical. This paper is about various medicinal benefits of fenugreek and its potential application as therapeutic agent against several diseases.


Origin of the conformational heterogeneity of cardiolipin-bound cytochrome C.
Interactions of cytochrome c (cyt c) with cardiolipin (CL) partially unfold the protein, activating its peroxidase function, a critical event in the execution of apoptosis. However, structural features of the altered protein species in the heterogeneous ensemble are difficult to probe with ensemble averaging. Analyses of the dye-to-heme distance distributions P(r) from time-resolved FRET (TR-FRET) have uncovered two distinct types of CL-bound cyt c conformations, extended and compact. We have combined TR-FRET, fluorescence correlation spectroscopy (FCS), and biolayer interferometry to develop a systematic understanding of the functional partitioning between the two conformations. The two subpopulations are in equilibrium with each other, with a submillisecond rate of conformational exchange reflecting the protein folding into a compact non-native state, as well as protein interactions with the lipid surface. Electrostatic interactions with the negatively charged lipid surface that correlate with physiologically relevant changes in CL concentrations strongly affect the kinetics of cyt c binding and conformational exchange. A predominantly peripheral binding mechanism, rather than deep protein insertion into the membrane, provides a rationale for the general denaturing effect of the CL surface and the large-scale protein unfolding. These findings closely relate to cyt c folding dynamics and suggest a general strategy for extending the time window in monitoring the kinetics of folding.


Friendship network characteristics and psychological well-being in late adolescence: exploring differences by gender and gender composition.
The aim of the present study was to examine the association between friendship networks and psychological well-being among 19-year-olds. The data used was a random sample of Swedish individuals born in 1990 who answered a questionnaire in 2009-2010. Friendship networks were considered in terms of three measures of emotional support. Six statements about the individual's emotional state were used to create a summary measure of psychological well-being. Gender and gender composition were included as potentially moderating factors. The association between friendship networks and psychological well-being was analysed by means of linear regression analysis (n = 1289). The results indicate that males' and females' friendship networks were similar with regard to quality and trust, whereas males' networks were characterized by less self-disclosure and a stronger preference for same-gender friendships. Gender composition did not matter for the support levels. Emotional support was associated with psychological well-being but there were gender differences: females seemed to benefit more health-wise from having high-quality (and trusting) networks. Moreover, whereas self-disclosure among males was positively linked to well-being, this was not the case among females. None of these associations were moderated by gender composition. In sum, friendship networks are beneficial for the psychological well-being among late adolescents, but there are some important differences according to gender.


Cholera toxin pretreatment protects against tumor necrosis factor lethality without compromising tumor response to therapy.
Antitumor therapy with tumor necrosis factor is limited by systemic toxic effects. We studied whether cholera toxin, a bacterial exotoxin that adenosine diphosphate-ribosylates the alpha-subunit of Gs proteins, could separate the lethal from the antitumor effects of tumor necrosis factor. A single dose of intravenous cholera toxin protected non-tumor-bearing mice from a lethal dose of Escherichia coli endotoxin administered 6 or 24 hours later. On the basis of these results, tumor-bearing mice were randomized to receive either cholera toxin or saline, followed 6 hours later by either human tumor necrosis factor (400 micrograms/kg) or saline. Tumor-bearing mice pretreated with cholera toxin had (1) reduced treatment-related mortality (0/11 vs 5/11 for saline controls) and (2) tumor regression similar to that of controls. In a separate experiment in tumor-bearing mice, intravenous human tumor necrosis factor treatment induced an increase in serum levels of murine tumor necrosis factor to a peak of 500 pg/mL at 1 hour in saline-pretreated controls, while a similar increase could not be detected in those mice pretreated with cholera toxin. These results suggest that pretreatment with cholera toxin can reduce the endogenous tumor necrosis factor response to administered tumor necrosis factor and separate the lethal from the antitumor effects. Cholera toxin may prove to be a useful tool for investigating the mechanisms underlying the varied effects of tumor necrosis factor.


Cancers attributable to excess body weight in Canada in 2010.
Excess body weight (body mass index [BMI] ≥ 25.00 kg/m2) is an established risk factor for diabetes, hypertension and cardiovascular disease, but its relationship to cancer is lesser-known. This study used population attributable fractions (PAFs) to estimate the cancer burden attributable to excess body weight in Canadian adults (aged 25+ years) in 2010. We estimated PAFs using relative risk (RR) estimates from the World Cancer Research Fund International Continuous Update Project, BMI-based estimates of overweight (25.00 kg/m2-29.99 kg/m2) and obesity (30.00+ kg/m2) from the 2000-2001 Canadian Community Health Survey, and cancer case counts from the Canadian Cancer Registry. PAFs were based on BMI corrected for the bias in self-reported height and weight. In Canada in 2010, an estimated 9645 cancer cases were attributable to excess body weight, representing 5.7% of all cancer cases (males 4.9%, females 6.5%). When limiting the analysis to types of cancer associated with high BMI, the PAF increased to 14.9% (males 17.5%, females 13.3%). Types of cancer with the highest PAFs were esophageal adenocarcinoma (42.2%), kidney (25.4%), gastric cardia (20.7%), liver (20.5%), colon (20.5%) and gallbladder (20.2%) for males, and esophageal adenocarcinoma (36.1%), uterus (35.2%), gallbladder (23.7%) and kidney (23.0%) for females. Types of cancer with the greatest number of attributable cases were colon (1445), kidney (780) and advanced prostate (515) for males, and uterus (1825), postmenopausal breast (1765) and colon (675) for females. Irrespective of sex or type of cancer, PAFs were highest in the Prairies (except Alberta) and the Atlantic region and lowest in British Columbia and Quebec. The cancer burden attributable to excess body weight is substantial and will continue to rise in the near future because of the rising prevalence of overweight and obesity in Canada.


Low-frequency oscillations employ a general coding of the spatio-temporal similarity of dynamic faces.
Brain networks use neural oscillations as information transfer mechanisms. Although the face perception network in occipitotemporal cortex is well-studied, contributions of oscillations to face representation remain an open question. We tested for links between oscillatory responses that encode facial dimensions and the theoretical proposal that faces are encoded in similarity-based "face spaces". We quantified similarity-based encoding of dynamic faces in magnetoencephalographic sensor-level oscillatory power for identity, expression, physical and perceptual similarity of facial form and motion. Our data show that evoked responses manifest physical and perceptual form similarity that distinguishes facial identities. Low-frequency induced oscillations (< 20Hz) manifested more general similarity structure, which was not limited to identity, and spanned physical and perceived form and motion. A supplementary fMRI-constrained source reconstruction implicated fusiform gyrus and V5 in this similarity-based representation. These findings introduce a potential link between "face space" encoding and oscillatory network communication, which generates new hypotheses about the potential oscillation-mediated mechanisms that might encode facial dimensions.


A synthesis of the theories and concepts of early human evolution.
Current evidence suggests that many of the major events in hominin evolution occurred in East Africa. Hence, over the past two decades, there has been intensive work undertaken to understand African palaeoclimate and tectonics in order to put together a coherent picture of how the environment of Africa has varied over the past 10 Myr. A new consensus is emerging that suggests the unusual geology and climate of East Africa created a complex, environmentally very variable setting. This new understanding of East African climate has led to the pulsed climate variability hypothesis that suggests the long-term drying trend in East Africa was punctuated by episodes of short alternating periods of extreme humidity and aridity which may have driven hominin speciation, encephalization and dispersals out of Africa. This hypothesis is unique as it provides a conceptual framework within which other evolutionary theories can be examined: first, at macro-scale comparing phylogenetic gradualism and punctuated equilibrium; second, at a more focused level of human evolution comparing allopatric speciation, aridity hypothesis, turnover pulse hypothesis, variability selection hypothesis, Red Queen hypothesis and sympatric speciation based on sexual selection. It is proposed that each one of these mechanisms may have been acting on hominins during these short periods of climate variability, which then produce a range of different traits that led to the emergence of new species. In the case of Homo erectus (sensu lato), it is not just brain size that changes but life history (shortened inter-birth intervals, delayed development), body size and dimorphism, shoulder morphology to allow thrown projectiles, adaptation to long-distance running, ecological flexibility and social behaviour. The future of evolutionary research should be to create evidence-based meta-narratives, which encompass multiple mechanisms that select for different traits leading ultimately to speciation.


Engineering a mevalonate pathway in Escherichia coli for production of terpenoids.
Isoprenoids are the most numerous and structurally diverse family of natural products. Terpenoids, a class of isoprenoids often isolated from plants, are used as commercial flavor and fragrance compounds and antimalarial or anticancer drugs. Because plant tissue extractions typically yield low terpenoid concentrations, we sought an alternative method to produce high-value terpenoid compounds, such as the antimalarial drug artemisinin, in a microbial host. We engineered the expression of a synthetic amorpha-4,11-diene synthase gene and the mevalonate isoprenoid pathway from Saccharomyces cerevisiae in Escherichia coli. Concentrations of amorphadiene, the sesquiterpene olefin precursor to artemisinin, reached 24 microg caryophyllene equivalent/ml. Because isopentenyl and dimethylallyl pyrophosphates are the universal precursors to all isoprenoids, the strains developed in this study can serve as platform hosts for the production of any terpenoid compound for which a terpene synthase gene is available.


Anti-ICAM-2 monoclonal antibody synergizes with intratumor gene transfer of interleukin-12 inhibiting activation-induced T-cell death.
Systemic treatment with an anti-ICAM-2 monoclonal antibody (mAb; EOL4G8) eradicates certain established mouse tumors through a mechanism dependent on the potentiation of a CTL-mediated response. However, well-established tumors derived from the MC38 colon carcinoma cell line were largely refractory to this treatment as well as to intratumor injection of a recombinant adenovirus encoding interleukin-12 (IL-12; AdCMVIL-12). We sought to design combined therapy strategies with AdCMVIL-12 plus anti-ICAM-2 mAbs and to identify their mechanism of action. Analysis of antitumor and toxic effects were performed with C57BL/6 mice bearing established MC38 tumors. Anti-ovalbumin T-cell receptor transgenic mice and tumors transfected with this antigen were used for in vitro and in vivo studies on activation-induced cell death (AICD) of CD8(+) T cells. Combined treatment with various systemic doses of EOL4G8 mAb plus intratumor injection of AdCMVIL-12 induced complete regression of MC38 tumors treated 7 days after implantation. Unfortunately, most of such mice succumbed to a systemic inflammatory syndrome that could be prevented if IFN-gamma activity were neutralized once tumors had been rejected. Importantly, dose reduction of EOL4G8 mAb opened a therapeutic window (complete cure of 9 of 18 cases without toxicity). We also show that ICAM-2 ligation by EOL4G8 mAb on activated CTLs prevents AICD, thus extending IFN-gamma production. Combination of intratumor gene transfer of IL-12and systemic anti-ICAM-2 mAb display synergistic therapeutic and toxic effects. CTL life extension resulting from AICD inhibition by anti-ICAM-2 mAbs is the plausible mechanism of action.


Determination of the T-DNA transfer and the T-DNA integration frequencies upon cocultivation of Arabidopsis thaliana root explants.
Using the Cre/lox recombination system, we analyzed the extent to which T-DNA transfer to the plant cell and T-DNA integration into the plant genome determine the transformation and cotransformation frequencies of Arabidopsis root cells. Without selection for transformation competence, the stable transformation frequency of shoots obtained after cocultivation and regeneration on nonselective medium is below 0.5%. T-DNA transfer and expression occur in 5% of the shoots, indicating that the T-DNA integrates in less than 10% of the transiently expressing plant cells. A limited fraction of root cells, predominantly located at the wounded sites and in the pericycle, are competent for interaction with agrobacteria and the uptake of a T-DNA, as demonstrated by histochemical GUS staining. When selection for transformation competence is applied, the picture is completely different. Then, approximately 50% of the transformants show transient expression of a second, nonselected T-DNA and almost 50% of these cotransferred T-DNAs are integrated into the plant genome. Our results indicate that both T-DNA transfer and T-DNA integration limit the transformation and cotransformation frequencies and that plant cell competence for transformation is based on these two factors.


Randomized controlled trial of EUS-guided fine needle aspiration techniques for the detection of malignant lymphadenopathy.
EUS-guided fine needle aspiration (EUS-FNA) is a highly accurate method of detecting malignant lymphadenopathy. The optimal methods for performing EUS-FNA to maximize sensitivity and to minimize the number of needle passes necessary are unknown. This is a report of the results of a prospective randomized controlled trial to determine the effect of suction, the site of FNA (edge or center of lymph node), and the method of preparation of cytologic specimens on accuracy, number of needle passes needed, and specimen quality. Consecutive patients with lymphadenopathy detected by EUS underwent FNA. Each lymph node was sampled with or without suction and from the edge or center in a 2 x 2 factorial design. The samples were expressed onto slides for cytology, and the residual material in the needle was analyzed by the cytospin-cellblock technique. Each aspirate was individually characterized for a diagnosis of malignancy, cellularity, and bloodiness. Forty-three patients with a total of 46 lymph nodes were evaluated. The final lymph node diagnosis was benign in 22 (48%), "suspicious for malignancy" in 6 (13%), and malignant in 18 (39%). The use of suction was associated with an increase in the cellularity of the specimen, but did not improve the likelihood of obtaining a correct diagnosis (OR 1.52: 95% CI [0.81, 2.85]). Samples obtained with suction were of worse quality because of excessive bloodiness (OR 4.7: 95% CI [1.99, 11.24]). Aspiration from the edge of the lymph node (compared with the center) did not increase the likelihood of a correct diagnosis (OR 1.16: 95% CI [0.42, 3.21]). For 78% of malignant lymph nodes, the correct diagnosis was obtained on the first needle pass and for 100% by the third pass. Cytospin-cellblock methods did not add any additional diagnostic information compared with direct smear cytology. The traditional method of applying suction during EUS-FNA does not improve diagnostic accuracy and worsens specimen bloodiness compared with FNA without suction. The site of FNA within the lymph node does not affect accuracy. When EUS-FNA is necessary, our recommendation is up to 3 FNAs without suction from the most convenient and safe location within abnormal-appearing lymph nodes.


Cardiotoxicity of cancer therapeutics: current issues in screening, prevention, and therapy.
In the context of modern cancer chemotherapeutics, cancer survivors are living longer and being exposed to potential comorbidities related to non-cancer side effects of such treatments. With close monitoring of cancer patients receiving potentially cardiotoxic medical therapies, oncologists, and cardiologists alike are identifying patients in both clinical and subclinical phases of cardiovascular disease related to such chemotherapies. Specifically, cardiotoxicity at the level of the myocardium and potential for the development of heart failure are becoming a growing concern with increasing survival of cancer patients. Traditional chemotherapeutic agents used commonly in the treatment of breast cancer and hematologic malignancies, such as anthracyclines and HER-2 antagonists, are well known to be associated with cardiovascular sequelae. Patients often present without symptoms and an abnormal cardiac imaging study performed as part of routine evaluation of patients receiving cardiotoxic therapies. Additionally, patients can present with signs and symptoms of cardiovascular disease months to years after receiving the chemotherapies. As the understanding of the physiology underlying the various cancers has grown, therapies have been developed that target specific molecules that represent key aspects of physiologic pathways responsible for cancer growth. Inhibition of these pathways, such as those involving tyrosine kinases, has lead to the potential for cardiotoxicity as well. In view of the potential cardiotoxicity of specific chemotherapies, there is a growing interest in identifying patients who are at risk of cardiotoxicity prior to becoming symptomatic or developing cardiotoxicity that may limit the use of potentially life-saving chemotherapy agents. Serological markers and novel cardiac imaging techniques have become the source of many investigations with the goal of screening patients for pre-clinical cardiotoxicity. Additionally, studies have been performed.


Canine leishmaniasis: the key points for qPCR result interpretation.
Diagnosis and follow up of CanL is difficult since the range of clinical signs is varied and seroprevalence is high in endemic areas. The aims of this study were: i) demonstrate the advantages of Leishmania qPCR to diagnose and control CanL and highlight its prognostic value and ii) propose guidelines for tissue selection and infection monitoring. This study included 710 dogs living in an endemic area of leishmaniasis. Forty percent (285/710) exhibited clinical signs consistent with CanL. Infection was detected in 36.3% (258/710) of the dogs of which 4.5% (32/710) were detected by qPCR, 16.2% (115/710) detected by ELISA and 15.6% (111/710) tested positive for both tests. Only 17.9% (127/710) of the dogs were classified sick (affected) with CanL. All symptomatic dogs with medium or high ELISA titers were qPCR-positive in blood samples. All dogs with inconclusive or low ELISA results with high or medium qPCR parasitemia values developed the disease. Seventy one percent of asymptomatic ELISA-positive dogs confirmed by qPCR (medium to high parasitemia) developed the disease. Bone marrow or lymph node aspirate should be selected to ensure the absence of the parasite in asymptomatic dogs: 100-1,000 parasites/ml in bone marrow are detectable in blood, whereas lower parasite loads are usually negative. Almost 10% of negative samples in blood were positive in conjunctival swabs. Because qPCR allows parasite quantification, it is an effective tool to confirm a diagnosis of CanL in (i) cases of inconclusive ELISA results, (ii) when the dog has not yet seroconverted, or (iii) for treatment monitoring.


Deciphering OPDA Signaling Components in the Momilactone-Producing Moss Calohypnum plumiforme.
Jasmonic acid (JA) and its biologically active form jasmonoyl-L-isoleucine (JA-Ile) regulate defense responses to various environmental stresses and developmental processes in plants. JA and JA-Ile are synthesized from α-linolenic acids derived from membrane lipids via 12-oxo-phytodienoic acid (OPDA). In the presence of JA-Ile, the COI1 receptor physically interacts with JAZ repressors, leading to their degradation, resulting in the transcription of JA-responsive genes by MYC transcription factors. Although the biosynthesis of JA-Ile is conserved in vascular plants, it is not recognized by COI1 in bryophytes and is not biologically active. In the liverwort Marchantia polymorpha, dinor-OPDA (dn-OPDA), a homolog of OPDA with two fewer carbons, and its isomer dn-iso-OPDA accumulate after wounding and are recognized by COI1 to activate downstream signaling. The moss Calohypnum plumiforme produces the antimicrobial-specialized metabolites, momilactones. It has been reported that JA and JA-Ile are not detected in C. plumiforme and that OPDA, but not JA, can induce momilactone accumulation and the expression of these biosynthetic genes, suggesting that OPDA or its derivative is a biologically active molecule in C. plumiforme that induces chemical defense. In the present study, we investigated the biological functions of OPDA and its derivatives in C. plumiforme. Searching for the components potentially involving oxylipin signaling from transcriptomic and genomic data revealed that two COI1, three JAZ, and two MYC genes were present. Quantification analyses revealed that OPDA and its isomer iso-OPDA accumulated in larger amounts than dn-OPDA and dn-iso-OPDA after wounding. Moreover, exogenously applied OPDA, dn-OPDA, or dn-iso-OPDA induced the transcription of JAZ genes. These results imply that OPDA, dn-OPDA, and/or their isomers potentially act as biologically active molecules to induce the signaling downstream of COI1-JAZ. Furthermore, co-immunoprecipitation analysis showed the physical interaction between JAZs and MYCs, indicating the functional conservation of JAZs in C. plumiforme with other plants. These results suggest that COI1-JAZ-MYC mediated signaling is conserved and functional in C. plumiforme.


Antioxidant cosupplementation therapy with vitamin C, vitamin E, and coenzyme Q10 in patients with oligoasthenozoospermia.
Overproduction of reactive oxygen species results in oxidative stress, a deleterious process that damages cell structure as well as lipids, proteins, and DNA. Oxidative stress plays a major role in various human diseases, such as oligoasthenozoospermia syndrome. We evaluated the effectiveness of antioxidant co-supplementation therapy using vitamin C, vitamin E, and coenzyme Q10 in men with oligoasthenozoospermia. Overall, 169 infertile men with oligoasthenozoospermia received antioxidant therapy with 80 mg/day vitamin C, 40 mg/day vitamin E, and 120 mg/day coenzyme Q10. We evaluated spermiogram parameters at baseline and at 3 and 6 months of follow-up. Significant improvements were evident in sperm concentration and motility following coenzyme Q10 therapy. Treatment resulted in 48 (28.4%) partner pregnancies, of which 16 (9.5%) were spontaneous. Significant improvements in sperm cell concentration and sperm motility were observed after 3 and 6 months of treatment. Vitamin C, vitamin E, and coenzyme Q10 supplementation resulted in a significant improvement in certain semen parameters. However, further studies are needed to empirically determine the effect of supplementation on pregnancy rate.


Orb and a long poly(A) tail are required for efficient oskar translation at the posterior pole of the Drosophila oocyte.
During Drosophila oogenesis, the posterior determinant, Oskar, is tightly localized at the posterior pole of the oocyte. The exclusive accumulation of Oskar at this site is ensured by localization-dependent translation of oskar mRNA: translation of oskar mRNA is repressed during transport and activated upon localization at the posterior cortex. Previous studies have suggested that oskar translation is poly(A)-independent. We show that a long poly(A) tail is required for efficient oskar translation, both in vivo and in vitro, but is not sufficient to overcome BRE-mediated repression. Moreover, we show that accumulation of Oskar activity requires the Drosophila homolog of Cytoplasmic Polyadenylation Element Binding protein (CPEB), Orb. As posterior localization of oskar mRNA is an essential prerequisite for its translation, it was critical to identify an allele of orb that does localize oskar mRNA to the posterior pole of the oocyte. We show that flies bearing the weak mutation orb(mel) localize oskar transcripts with a shortened poly(A) that fails to enhance oskar translation, resulting in reduced Oskar levels and posterior patterning defects. We conclude that Orb-mediated cytoplasmic polyadenylation stimulates oskar translation to achieve the high levels of Oskar protein necessary for posterior patterning and germline differentiation.


Nutritional Risk in Cancer Patients 65 and Older Undergoing Systemic Phase I Treatment.
Malnutrition is common in cancer patients and recognized as an important component of adverse outcomes, including increased morbidity, mortality, and decreased quality of life (QOL). Quality of life is an overarching term for quality of various domains of life. It is a standard level that consists of the expectations of an individual for a good life. These expectations are guided by values, goals, and sociocultural context. It is a subjective, multidimensional concept defining a standard level for emotional, physical, material, and social well-being (Bottomley, 2012). Nutritional risk is not consistently assessed in the older adult cancer patient population. The purpose of this secondary analysis was to identify variables related to nutritional risk in the cancer patient 65 years and older receiving systemic treatments. The study described the relationship between nutritional risk and four domains of QOL (physical, social, emotional, and functional). A sample of 73 patients was accrued for this study from an NCI-funded RO1 aimed at integrating supportive care for cancer patients. The Mini Nutritional Assessment-Short Form instrument was used to assess for nutritional risk. Findings revealed the strongest correlation with nutritional risk was BMI status (r = .47, p < .0001). Multiple regression analysis demonstrated factors associated with nutritional risk included BMI, previous chemotherapy, and physical subscale of the Functional Assessment of Cancer Therapy-General QOL instrument. Descriptive data reinforced the importance of assessment and intervention to support nutritional status. Nutrition impacts all dimensions of QOL and is even more important in an aging population. Advanced practitioners can contribute greatly to advancing this area of practice.


Inborn-like errors of metabolism are determinants of breast cancer risk, clinical response and survival: a study of human biochemical individuality.
Breast cancer remains a leading cause of morbidity and mortality worldwide yet methods for early detection remain elusive. We describe the discovery and validation of biochemical signatures measured by mass spectrometry, performed upon blood samples from patients and controls that accurately identify (>95%) the presence of clinical breast cancer. Targeted quantitative MS/MS conducted upon 1225 individuals, including patients with breast and other cancers, normal controls as well as individuals with a variety of metabolic disorders provide a biochemical phenotype that accurately identifies the presence of breast cancer and predicts response and survival following the administration of neoadjuvant chemotherapy. The metabolic changes identified are consistent with inborn-like errors of metabolism and define a continuum from normal controls to elevated risk to invasive breast cancer. Similar results were observed in other adenocarcinomas but were not found in squamous cell cancers or hematologic neoplasms. The findings describe a new early detection platform for breast cancer and support a role for pre-existing, inborn-like errors of metabolism in the process of breast carcinogenesis that may also extend to other glandular malignancies. Statement of Significance: Findings provide a powerful tool for early detection and the assessment of prognosis in breast cancer and define a novel concept of breast carcinogenesis that characterizes malignant transformation as the clinical manifestation of underlying metabolic insufficiencies.


Phylogeography of a Tertiary relict plant, Meconopsis cambrica (Papaveraceae), implies the existence of northern refugia for a temperate herb.
The perennial herb Meconopsis cambrica, a western European endemic, is the only European species of the otherwise Himalayan genus Meconopsis and has been interpreted as a Tertiary relict species. Using rbcL and ITS sequence variation, we date the split between M. cambrica and its sister clade Papaver s.str. to the Middle to Upper Miocene (12.8 Myr, 6.4-19.2 Myr HPD). Within M. cambrica, cpDNA sequence variation reveals the existence of two groups of populations with a comparable level of genetic variation: a northern group from Great Britain, the Massif Central, the western Pyrenees and the Iberian System, and a southern group from the central and eastern Pyrenees. Populations from the Cantabrian Mountains were placed in both groups. Based on ITS sequence variation, the divergence between these two groups can be dated to 1.5 Myr (0.4-2.8 Myr HPD), and the age of the British populations is estimated as 0.37 Myr (0.0-0.9 Myr HPD). Amplified fragment length polymorphism results confirm the distinctive nature of the populations from Britain, the Massif Central and the central and eastern Pyrenees. These patterns of latitudinal variation of M. cambrica differ from patterns of longitudinal differentiation found in many other temperate species and imply glacial survival of the northern populations in northerly refugia. The primary differentiation into northern and southern cpDNA groups dates to near the onset of the Quaternary and suggests that an ancient phylogeographic pattern has survived through several glacial periods. Our data provide evidence that the species has persisted for a long period with a highly fragmented and probably very localized distribution.


Characterizing the role of disparity information in alleviating visual crowding.
The ability to identify a target is reduced by the presence of nearby objects, a phenomenon known as visual crowding. The extent to which crowding impairs our perception is generally governed by the degree of similarity between a target stimulus and its surrounding flankers. Here we investigated the influence of disparity differences between target and flankers on crowding. Orientation discrimination thresholds for a parafoveal target were first measured when the target and flankers were presented at the same depth to establish a flanker separation that induced a significant elevation in threshold for each individual. Flankers were subsequently fixed at this spatial separation while the disparity of the flankers relative to the target was altered. For all participants, thresholds showed a systematic decrease as flanker-target disparity increased. The resulting tuning function was asymmetric: Crowding was lower when the target was perceived to be in front of the flankers rather than behind. A series of control experiments confirmed that these effects were driven by disparity, as opposed to other factors such as flanker-target separation in three-dimensional (3-D) space or monocular positional offsets used to create disparity. When flankers were distributed over a range of crossed and uncrossed disparities, such that the mean was in the plane of the target, there was an equivalent or greater release of crowding compared to when all flankers were presented at the maximum disparity of that range. Overall, our results suggest that depth cues can reduce the effects of visual crowding, and that this reduction is unlikely to be caused by grouping of flankers or positional shifts in the monocular image.


Identification of a contact site for residue 19 of parathyroid hormone (PTH) and PTH-related protein analogs in transmembrane domain two of the type 1 PTH receptor.
Recent functional studies have suggested that position 19 in PTH interacts with the portion of the PTH-1 receptor (P1R) that contains the extracellular loops and seven transmembrance helices (TMs) (the J domain). We tested this hypothesis using the photoaffinity cross-linking approach. A PTHrP(1-36) analog and a conformationally constrained PTH(1-21) analog, each containing para-benzoyl-l-phenylalanine (Bpa) at position 19, each cross-linked efficiently to the P1R expressed in COS-7 cells, and digestive mapping analysis localized the cross-linked site to the interval (Leu232-Lys240) at the extracellular end of TM2. Point mutation analysis identified Ala234, Val235, and Lys240 as determinants of cross-linking efficiency, and the Lys240-->Ala mutation selectively impaired the binding of PTH(1-21) and PTH(1-19) analogs, relative to that of PTH(1-15) analogs. The findings support the hypothesis that residue 19 of the receptor-bound ligand contacts, or is close to, the P1R J domain-specifically, Lys240 at the extracellular end of TM2. The findings also support a molecular model in which the 1-21 region of PTH binds to the extracellular face of the P1R J domain as an alpha-helix.


Evaluation of Jeffamine® Core PAMAM Dendrimers for Simultaneous Removal of Divalent Heavy Metal Ions from Aqueous Solutions by Polymer Assisted Ultrafiltration.
Different generations (G3-G4) of amine-terminated Jeffamine® T-403 core poly(amidoamine) PAMAM dendrimers (JCPDs) were used as new macromolecular heavy metal chelating agent templates in polymer assisted ultrafiltration (PAUF) for the investigation of their removal ability for some of the divalent metal ions: Cu, Co, Ni, Cd, and Zn from aqueous solutions under competitive conditions. The effects of pH and generation size of JCPDs were also investigated. Extent of binding (EOB) data can be appropriately expressed by a tetradentate coordination for JCPDs at pH 9 where the maximum removal of metal ions was observed. At pH 9.0, the affinity of both generations towards heavy metal ions was also observed in the decreasing order of Zn(II) > Co(II) > Ni(II) > Cu(II) > Cd(II). Results revealed that the highest total binding capacity was observed for G3-NH2 (262.79 ± 1.62 mg/g) as a little bit higher than that of G4-NH2 (257.27 ± 2.57 mg/g). EOB studies also proved the active contribution of amide groups to metal binding ability of PAMAMs. Both generations were selective towards Cu(II) ions at lower pH 5 and pH 7. From these results, it was concluded that studied JCPDs have the desired technical properties to be used for the removal of toxic metals from wastewaters.


Rapid eye movement sleep and hippocampal theta oscillations precede seizure onset in the tetanus toxin model of temporal lobe epilepsy.
Improved understanding of the interaction between state of vigilance (SOV) and seizure onset has therapeutic potential. Six rats received injections of tetanus toxin (TeTX) in the ventral hippocampus that resulted in chronic spontaneous seizures. The distribution of SOV before 486 seizures was analyzed for a total of 19 d of recording. Rapid eye movement sleep (REM) and exploratory wake, both of which express prominent hippocampal theta rhythm, preceded 47 and 34%, for a total of 81%, of all seizures. Nonrapid eye movement sleep (NREM) and nonexploratory wake, neither of which expresses prominent theta, preceded 6.8 and 13% of seizures. We demonstrate that identification of SOV yields significant differentiation of seizure susceptibilities, with the instantaneous seizure rate during REM nearly 10 times higher than baseline and the rate for NREM less than half of baseline. Survival analysis indicated a shorter duration of preseizure REM bouts, with a maximum transition to seizure at ∼90 s after the onset of REM. This study provides the first analysis of a correlation between SOV and seizure onset in the TeTX model of temporal lobe epilepsy, as well as the first demonstration that hippocampal theta rhythms associated with natural behavioral states can serve a seizure-promoting role. Our findings are in contrast with previous studies suggesting that the correlations between SOV and seizures are primarily governed by circadian oscillations and the notion that hippocampal theta rhythms inhibit seizures. The documentation of significant SOV-dependent seizure susceptibilities indicates the potential utility of SOV and its time course in seizure prediction and control.


Biomarker-guided preemption of steroid-refractory graft-versus-host disease with α-1-antitrypsin.
Steroid-refractory (SR) acute graft-versus-host disease (GVHD) remains a major cause of nonrelapse mortality (NRM) after allogeneic hematopoietic cell transplantation (HCT), but its occurrence is not accurately predicted by pre-HCT clinical risk factors. The Mount Sinai Acute GVHD International Consortium (MAGIC) algorithm probability (MAP) identifies patients who are at high risk for developing SR GVHD as early as 7 days after HCT based on the extent of intestinal crypt damage as measured by the concentrations of 2 serum biomarkers, suppressor of tumorigenesis 2 and regenerating islet-derived 3α. We conducted a multicenter proof-of-concept "preemptive" treatment trial of α-1-antitrypsin (AAT), a serine protease inhibitor with demonstrated activity against GVHD, in patients at high risk for developing SR GVHD. Patients were eligible if they possessed a high-risk MAP on day 7 after HCT or, if initially low risk, became high risk on repeat testing at day 14. Thirty high-risk patients were treated with twice-weekly infusions of AAT for a total of 16 doses, and their outcomes were compared with 90 high-risk near-contemporaneous MAGIC control patients. AAT treatment was well tolerated with few toxicities, but it did not lower the incidence of SR GVHD compared with controls (20% vs 14%, P = .56). We conclude that real-time biomarker-based risk assignment is feasible early after allogeneic HCT but that this dose and schedule of AAT did not change the incidence of SR acute GVHD. This trial was registered at www.clinicaltrials.gov as #NCT03459040.