Incidence and potential implications of the toxic metabolite methylglyoxal in cell culture: A review.
Methylglyoxal is a toxic metabolite unavoidably produced in mammalian systems as a by-product of glycolysis. Detoxification of this compound occurs principally through the glyoxalase pathway, which consists of glyoxalase I and glyoxalase II, and requires reduced glutathione as a co-enzyme. Recently, it has been demonstrated that variations in glucose, glutamine and fetal bovine serum levels can cause significant changes in the intracellular concentration of methylglyoxal. More importantly, comparative studies involving wild-type Chinese hamster ovary cells and clones overexpressing glyoxalase I indicate that glucose and glutamine, within the range normally found in cell culture media, can cause decreased cell viability mediated solely through increased production of methylglyoxal. In addition, endogenously produced methylglyoxal has been shown to cause apoptosis in cultured HL60 cells. While the exact mechanism of the impact of methylglyoxal on cultured cells is unknown, methylglyoxal is a potent protein and nucleic acid modifying agent at physiological concentrations and under physiological conditions. Protein modification occurs mainly at arginine, lysine and cysteine residues and is believed to be an important signal for the degradation of senescent proteins. Modification of arginine and lysine results in the irreversible formation of advanced glycation endproducts, whereas modification of cysteine results in the formation of a highly reversible hemithioacetal. Methylglyoxal also forms adducts with nucleic acids, principally with guanyl residues. At high extracellular concentrations, it is genotoxic to cells grown in culture. Even at physiological concentrations (100 nM free methylglyoxal), methylglyoxal can modify unprotected plasmid DNA and cause gene mutation and abnormal gene expression.


Modification of nasal epithelial potential differences of individuals with cystic fibrosis consequent to local administration of a normal CFTR cDNA adenovirus gene transfer vector.
Mutations in the cystic fibrosis transmembrane conductance regulator gene (CFTR) manifest on the nasal epithelial surface of individuals with cystic fibrosis (CF) by Na+ hyperabsorption and diminished beta-agonist-induced Cl- conductance, resulting in an abnormal bioelectric phenotype across the nasal epithelium. A clinical trial was conducted to determine if a replication-deficient, recombinant adenovirus vector containing a normal copy of the CFTR cDNA (AdCFTR) could, when administered to the nasal epithelium, correct the abnormal bioelectric phenotype. Nine individuals with CF received 2 x 10(5) to 2 x 10(8.5) plaque forming units of AdCFTR to the epithelium of one nostril. Measurements made included: baseline electrical potential difference (PD) between the surface of the nasal epithelium and the interstitial fluid, change in PD in response to amiloride, which inhibits apical Na+ channels, and change in PD in response to isoproterenol in a low Cl- solution, a measure of cAMP-regulated Cl- conductance. The functional integrity of the epithelium was evaluated by the PD response to ATP. Each individual served as their own control with measurements made in the nostril to be treated before AdCFTR administration, and in the contralateral untreated nostril. On the average, in the treated nostril over 2 weeks after the local administration of the adenovirus vector compared to measurements made in the same nostril before treatment, baseline PD decreased toward normal (-53.3 +/- 4.0 to -34.6 +/- 3.4, p = 0.01), response to amiloride decreased toward normal (36.9 +/- 4.7 to 19.7 +/- 3.0, p = 0.02), and response to low Cl- and isoproterenol increased toward normal (-4.5 +/- 1.5 to -9.1 +/- 2.1, p = 0.05). There were no changes in response to ATP (-15.3 +/- 2.7 to -15.8 +/- 1.9, p = 0.39), suggesting that the epithelium remained functionally intact. Importantly, there were no significant changes in measurements made in the untreated nostril. While limited to the nasal epithelium, these data suggest an adenovirus vector can safely deliver sufficient CFTR cDNA function to improve the abnormal CF bioelectric phenotype.


Spin canting across core/shell Fe3O4/MnxFe3-xO4 nanoparticles.
Magnetic nanoparticles (MNPs) have become increasingly important in biomedical applications like magnetic imaging and hyperthermia based cancer treatment. Understanding their magnetic spin configurations is important for optimizing these applications. The measured magnetization of MNPs can be significantly lower than bulk counterparts, often due to canted spins. This has previously been presumed to be a surface effect, where reduced exchange allows spins closest to the nanoparticle surface to deviate locally from collinear structures. We demonstrate that intraparticle effects can induce spin canting throughout a MNP via the Dzyaloshinskii-Moriya interaction (DMI). We study ~7.4 nm diameter, core/shell Fe3O4/MnxFe3-xO4 MNPs with a 0.5 nm Mn-ferrite shell. Mössbauer spectroscopy, x-ray absorption spectroscopy and x-ray magnetic circular dichroism are used to determine chemical structure of core and shell. Polarized small angle neutron scattering shows parallel and perpendicular magnetic correlations, suggesting multiparticle coherent spin canting in an applied field. Atomistic simulations reveal the underlying mechanism of the observed spin canting. These show that strong DMI can lead to magnetic frustration within the shell and cause canting of the net particle moment. These results illuminate how core/shell nanoparticle systems can be engineered for spin canting across the whole of the particle, rather than solely at the surface.


Scaling of entropy production under coarse graining in active disordered media.
Entropy production plays a fundamental role in the study of nonequilibrium systems by offering a quantitative handle on the degree of time-reversal symmetry breaking. It depends crucially on the degree of freedom considered as well as on the scale of description. How the entropy production at one resolution of the degrees of freedom is related to the entropy production at another resolution is a fundamental question which has recently attracted interest. This relationship is of particular relevance to coarse-grained and continuum descriptions of a given phenomenon. In this work, we derive the scaling of the entropy production under iterative coarse graining on the basis of the correlations of the underlying microscopic transition rates for noninteracting particles in active disordered media. Our approach unveils a natural criterion to distinguish equilibrium-like and genuinely nonequilibrium macroscopic phenomena based on the sign of the scaling exponent of the entropy production per mesostate.


The intrinsic value of choice: The propensity to under-delegate in the face of potential gains and losses.
Human beings are often faced with a pervasive problem: whether to make their own decision or to delegate the decision task to someone else. Here, we test whether people are inclined to forgo monetary rewards in order to retain agency when faced with choices that could lead to losses and gains. In a simple choice task, we show that participants choose to pay in order to control their own payoff more than they should if they were to maximize monetary rewards and minimize monetary losses. This tendency cannot be explained by participants' overconfidence in their own ability, as their perceived ability was elicited and accounted for. Nor can the results be explained by lack of information. Rather, the results seem to reflect an intrinsic value for choice, which emerges in the domain of both gains and of losses. Moreover, our data indicate that participants are aware that they are making suboptimal choices in the normative sense, but do so anyway, presumably for psychological gains.


Yersinia pseudotuberculosis and Yersinia pestis are more resistant to bactericidal cationic peptides than Yersinia enterocolitica.
The action of bactericidal polycationic peptides was compared in Yersinia spp. by testing peptide binding to live cells and changes in outer membrane (OM) morphology and permeability. Moreover, polycation interaction with LPS was studied by measuring the dependence of dansylcadaverine displacement and zeta potential on polycation concentration. When growth at 37 degrees C, Yersinia pestis and Yersinia pseudotuberculosis bound less polymyxin B (PMB) than pathogenic or non-pathogenic Yersinia enterocolitica, regardless of virulence plasmid expression. Y. pseudotuberculosis OMs were unharmed by PMB concentrations causing extensive OM blebbing in Y. enterocolitica. The permeability to lysozyme caused by PMB was greater in Y. enterocolitica than in Y. pseudotuberculosis or Y. pestis and differences increased at 37 degrees C. Similar observations were made with other polycations using a polymyxin/novobiocin permeability assay. With LPS of cells grown at 26 degrees C, polycation binding was highest for Y. pseudotuberculosis and lowest for Y. pestis, with Y. enterocolitica yielding intermediate results which were lower for pathogenic than for non-pathogenic strains. With LPS of cells grown at 37 degrees C, polycation binding remained unchanged for Y. pestis and pathogenic Y. enterocolitica, increased for non-pathogenic Y. enterocolitica and decreased for Y. pseudotuberculosis to Y. pestis levels. Polycation binding related in part to differences in charge density (zeta potential) of LPS aggregates, suggesting similar effects at bacterial surfaces. It is suggested that species and temperature differences in polycation resistance relate to infection route, invasiveness and intracellular multiplication of Yersinia spp.


Robust phase-unwrapping algorithm with a spatial binary-tree image decomposition.
The search for fast and robust phase-unwrapping algorithms remains an important problem in the development of real-time interferometric systems. Our phase-unwrapping approach uses a spatial binary-tree image decomposition to permit maximum parallelism in implementation. At each node in the tree structure, a single unwrapping decision is made between two image blocks. The unwrapping rule is derived from a statistical-estimation framework. Specifically, a maximum-likelihood estimate of the demodulation term is used. This term can be viewed as that which minimizes a discontinuity-penalizing cost function. We show that the algorithm exhibits a high level of robustness. Quantitative measures of performance are provided, and many phase maps are shown for subjective evaluation.


Adverse Events of Oncologic Immunotherapy and Their Management.
Over the past two decades, immunotherapy has emerged as a promising treatment option for patients with cancer. However, newer versions of immunotherapy, such as checkpoint inhibitors, may be associated with unusual adverse effects (AEs) that can range in severity from mild to life-threatening. Unlike common AEs of conventional chemotherapy, which have a predictable nadir or cyclic pattern after administration, AEs of these newer immunotherapies are variable, depending on the type of immunotherapy, route of administration, and mechanism of action. The onset and resolution of these AEs may be present at any time, during administration of treatment, a few weeks after administration of treatment, or several months after completion of treatment. Therefore, improving outcomes in patients undergoing oncologic immunotherapy requires oncology nurses' knowledge and understanding of various immunotherapy agents, as well as early recognition and management of potential AEs, especially AEs associated with checkpoint inhibitors and other therapies that manipulate T-cell activation causing autoimmune toxicity. This article draws upon current evidence from systematic reviews, meta-analyses, and expert consensus guidelines to provide a brief overview of common immunotherapies used in cancer and management of their associated AEs.


Environmental abundance of Anopheles (Diptera: Culicidae) larval habitats on land cover change sites in Karima Village, Mwea Rice Scheme, Kenya.
A study was carried out at Karima Village in the Mwea Rice Irrigation Scheme in Kenya to assess the impact of rice husbandry and associated land cover change for mosquito larval abundance. A multi-temporal, land use land cover (LULC) classification dataset incorporating distributions of Anopheles arabiensis aquatic larval habitats was produced in ERDAS Imagine version 8.7 using combined images from IKONOS at 4m spatial resolution from 2005 and Landsat Thematic Mapper (TM)trade mark classification data at 30-meters spatial resolution from 1988 for Karima. Of 207 larval habitats sampled, most were either canals (53.4%) or paddies (45.9%), and only one habitat was classified as a seep (0.5%). The proportion of habitats that were poorly drained was 55.1% compared with 44.9% for the habitats that were well drained. An LULC base map was generated. A grid incorporating each rice paddy was overlaid over the LULC maps stratifying each cell based on levels of irrigation. Paddies/grid cells were classified as 1) well irrigated and 2) poorly irrigated. Early stages of rice growth showed peak larval production during the early part of the cropping cycle (rainy season). Total LULC change for Karima over 16 years was 59.8%. Of those areas in which change was detected, the LULC change for Karima was 4.30% for rice field to built environment, 8.74% for fallow to built environment, 7.19% for rice field to fallow, 19.03% built to fallow, 5.52% for fallow to rice field, and 8.35% for built environment to rice field. Of 207 aquatic habitats in Karima, 54.1 (n = 112) were located in LULC change sites and 45.9 (n = 95) were located in LULC non-change sites. Rice crop LULC maps derived from IKONOS and TM data in geographic information systems can be used to investigate the relationship between rice cultivation practices and higher anopheline larval habitat distribution.


Crystal structure of a genomically encoded fosfomycin resistance protein (FosA) at 1.19 A resolution by MAD phasing off the L-III edge of Tl(+).
The fosfomycin resistance protein (FosA) catalyzes the Mn(II)- and K+-dependent addition of glutathione to the oxirane of the antibiotic fosfomycin. The crystal structure of FosA from Pseudomonas aeruginosa was solved at a resolution of 1.19 A by multiwavelength anomalous diffraction at the L-III edge of a Tl+ derivative. The structure solution took advantage of the ability of Tl+ to substitute for K+. The existence of multiple Tl sites in the asymmetric unit suggests that this may be a generally useful technique for phasing protein crystal structures. A 1.35 A resolution structure with phosphate bound in the active site shows that the Mn(II) center has a rare four-coordinate geometry. The structure of the fosfomycin complex at 1.19 A resolution indicates that the Mn(II) center is close to five-coordinate with trigonal bipyramidal geometry and a ligand set consisting of two histidines (H7 and H64) and one phosphonate oxygen occupying the equatorial sites and the carboxylate of E110 at one of the apical sites. The oxirane oxygen of the substrate is located at the other apical site but is 0.2 A beyond the average Mn-O distance for five-coordinate Mn(II). The Mn(II) center is proposed to stabilize the alkoxide in the transition state, while the nearby hydroxyl group of T9 acts as a proton donor in the reaction. The K+ ion located 6.5 A from the Mn(II) appears to help orient the substrate for nucleophilic attack.


Sumatriptan (rectal route of administration) for acute migraine attacks in adults.
Migraine is a highly disabling condition for the individual and also has wide-reaching implications for society, healthcare services, and the economy. Sumatriptan is an abortive medication for migraine attacks, belonging to the triptan family. Rectal administration may be preferable to oral for individuals experiencing nausea and/or vomiting. To determine the efficacy and tolerability of rectal sumatriptan compared to placebo and other active interventions in the treatment of acute migraine attacks in adults. We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, online databases, and reference lists for studies through 13 October 2011. We included randomised, double-blind, placebo- and/or active-controlled studies using rectally administered sumatriptan to treat a migraine headache episode, with at least 10 participants per treatment arm. Two review authors independently assessed trial quality and extracted data. We used numbers of participants achieving each outcome to calculate relative risk (or 'risk ratio') and numbers needed to treat to benefit (NNT) or harm (NNH) compared to placebo or a different active treatment. Three studies (866 participants) compared rectally administered sumatriptan with placebo or an active comparator. Most of the data were for the 12.5 mg and 25 mg doses. For the majority of efficacy outcomes, sumatriptan surpassed placebo. For sumatriptan 12.5 mg versus placebo the NNTs were 5.2 and 3.2 for headache relief at one and two hours, respectively. Results for the 25 mg dose were similar to the 12.5 mg dose, and there were no significant differences between the two doses for any of the outcomes analysed. The NNTs for sumatriptan 25 mg versus placebo were 4.2, 3.2, and 2.4 for pain-free at two hours, headache relief at one hour, and headache relief at two hours, respectively.Relief of functional disability was greater with sumatriptan than with placebo, with NNTs of 8.0 and 4.0 for the 12.5 mg and 25 mg doses, respectively. For the most part, adverse events were transient and mild and were more common with sumatriptan than with placebo, but there were insufficient data to perform any analyses.Direct comparison of sumatriptan with active treatments was limited to one study comparing sumatriptan 25 mg with ergotamine tartrate 2 mg + caffeine 100 mg. Based on limited amounts of data, sumatriptan 25 mg, administered rectally, is an effective treatment for acute migraine attacks, with participants in these studies experiencing a significant reduction in headache pain and functional disability within two hours of treatment. The lack of data on relief of headache-associated symptoms or incidence of adverse events limits any conclusions that can be drawn.


uORF-mediated translational control: recently elucidated mechanisms and implications in cancer.
Protein synthesis is tightly regulated, and its dysregulation can contribute to the pathology of various diseases, including cancer. Increased or selective translation of mRNAs can promote cancer cell proliferation, metastasis and tumor expansion. Translational control is one of the most important means for cells to quickly adapt to environmental stresses. Adaptive translation involves various alternative mechanisms of translation initiation. Upstream open reading frames (uORFs) serve as a major regulator of stress-responsive translational control. Since recent advances in omics technologies including ribo-seq have expanded our knowledge of translation, we discuss emerging mechanisms for uORF-mediated translation regulation and its impact on cancer cell biology. A better understanding of dysregulated translational control of uORFs in cancer would facilitate the development of new strategies for cancer therapy.


Fragmentation of methyl chloride studied by partial positive and negative ion-yield spectroscopy.
The authors present partial-ion-yield experiments on the methyl chloride molecule excited in the vicinity of the Cl2p and C1s inner shells. A large number of fragments, cations produced by dissociation or recombination processes, as well as anionic species, have been detected. Although the spectra exhibit different intensity distributions depending on the core-excited atom, general observations include strong site-selective fragmentation along the C-Cl bond axis and a strong intensity dependence of transitions involving Rydberg series on fragment size.


Efficacy and toxicity of estrogens commonly used to terminate canine pregnancy.
Three groups of bitches were treated with diethylstilbestrol (75 micrograms/kg) orally for 7 days (n = 12), estradiol cypionate intramuscularly once (22 micrograms/kg; n = 12), or estradiol cypionate intramuscularly once (44 micrograms/kg; n = 12). Treatments commenced during late proestrus (n = 4/group), the fourth day of behavioral estrus (n = 4/group), or the second day of diestrus (n = 4/group). All bitches were bred on alternate days throughout estrus to stud dogs of known fertility. Ovariohysterectomies were performed on day 25 of diestrus to diagnose pregnancy and to assess any pathologic changes in the uterus. Eleven bitches treated with diethylstilbestrol, 6 bitches treated with the low dosage of estradiol cypionate, and 4 bitches receiving the high dosage of estradiol cypionate were pregnant at the time of surgery. Ten of the bitches treated with estrogens during proestrus, 6 treated during estrus, and 4 treated during diestrus were pregnant. The serum concentration of progesterone in 2 bitches treated with the high dosage of estradiol cypionate decreased to less than 2 ng/ml by day 25 of diestrus, suggesting premature luteal regression. Diethylstilbestrol appeared to have little efficacy in terminating pregnancy. Estradiol cypionate appeared to have greater efficacy when administered during estrus or early diestrus; however, pyometra developed in 2 bitches treated with this estrogen during diestrus.


Sequential Combination Therapy of CDK Inhibition and Doxorubicin Is Synthetically Lethal in p53-Mutant Triple-Negative Breast Cancer.
Triple-negative breast cancer (TNBC) is an aggressive malignancy in which the tumors lack expression of estrogen receptor, progesterone receptor, and HER2. Hence, TNBC patients cannot benefit from clinically available targeted therapies and rely on chemotherapy and surgery for treatment. While initially responding to chemotherapy, TNBC patients are at increased risk of developing distant metastasis and have decreased overall survival compared with non-TNBC patients. A majority of TNBC tumors carry p53 mutations, enabling them to bypass the G1 checkpoint and complete the cell cycle even in the presence of DNA damage. Therefore, we hypothesized that TNBC cells are sensitive to cell-cycle-targeted combination therapy, which leaves nontransformed cells unharmed. Our findings demonstrate that sequential administration of the pan-CDK inhibitor roscovitine before doxorubicin treatment is synthetically lethal explicitly in TNBC cells. Roscovitine treatment arrests TNBC cells in the G2-M cell-cycle phase, priming them for DNA damage. Combination treatment increased frequency of DNA double-strand breaks, while simultaneously reducing recruitment of homologous recombination proteins compared with doxorubicin treatment alone. Furthermore, this combination therapy significantly reduced tumor volume and increased overall survival compared with single drug or concomitant treatment in xenograft studies. Examination of isogenic immortalized human mammary epithelial cells and isogenic tumor cell lines found that abolishment of the p53 pathway is required for combination-induced cytotoxicity, making p53 a putative predictor of response to therapy. By exploiting the specific biologic and molecular characteristics of TNBC tumors, this innovative therapy can greatly impact the treatment and care of TNBC patients. Mol Cancer Ther; 15(4); 593-607. ©2016 AACR.


Expression of the retinoblastoma-related p107 and Rb2/p130 genes in human placenta: an immunohistochemical study.
It has been proposed that tumor suppressor genes may have a role in the mechanisms of proliferation and differentiation during human placental development. The Retinoblastoma gene family is a well known family of tumor suppressor genes. Many studies have pointed out a role of this family not only in cell cycle progression, but also during development and differentiation. On the light of these observations we have investigated the immunohistochemical expression pattern of the Retinoblastoma family members, p107 and Rb2/p130 in human placenta samples in first trimester and full-term placental sections. p107 and pRb2/p130 showed the most abundant expression levels during the first trimester of gestation and progressively declined to being barely detectable in the placenta by late gestation. These results indicate that the expression of the above genes is modulated during placental development and suggest a mechanism for controlling trophoblast proliferation.


Risk mitigation for travelers: managing endemic and emerging threats.
International travel continues to increase in numbers and complexity. Despite the availability of pretravel health advice, travelers remain at risk for exposure to common organisms as well as emerging pathogens. With low rates of travel clinic utilization, it is important for the general practitioner to remain aware of the importance of travel health, risk factors unique to individual patients, and evolving threats. This review highlights important considerations when evaluating ill travelers, incorporating emerging infectious threats.


In vivo imaging of T cell priming.
The rules by which naïve T cells decide whether and how to respond to antigenic stimuli are incompletely understood. Using multiphoton intravital microscopy (MP-IVM) in lymph nodes (LNs), we have shown that CD8+ T cells are primed by antigen-presenting dendritic cells (DCs) in three consecutive phases. During phase one, T cells undergo brief serial contacts with many DCs for several hours after homing into the LNs. Subsequently, during phase two, T cells engage in prolonged stable interactions with DCs. Finally, in the third phase, T cells return to transient interactions with DCs as they begin to proliferate and eventually leave the LNs. We have examined the influence of antigen dose on the duration of phase one by systematically varying both the number of cognate peptide-major histocompatability (pMHC) complexes per DC and the density of cognate pMHC complex-presenting DCs per LN. The duration of phase one and the kinetics of CD8+ T cell activation were inversely correlated with both parameters. Very few pMHC complexes were needed for full T cell activation and effector differentiation. Furthermore, there was a sharp threshold of antigen dose below which T cells did not transition to phase two but continued to migrate until they exited the LN, unactivated. The stability of peptide binding to MHC was a critical determinant of this threshold antigen dose in vivo. Our results suggest an integrative mechanism that allows T cells to reach an informed decision about whether to respond, based on the overall antigen dose encountered.


Role of systemic chemotherapy in urothelial urinary bladder cancer.
Radical cystectomy is the standard of care for patients with localized muscle-invasive bladder cancer; however, 50 percent of patients still relapse in distant sites following surgery. A systemic approach is needed to improve outcomes in bladder cancer in the metastatic and perioperative settings. We reviewed the literature for use of systemic chemotherapy in bladder cancer and its role in metastatic, neoadjuvant, and adjuvant settings, including patients with comorbidities and renal dysfunction. Current controversies on the role of chemotherapy in neoadjuvant and adjuvant settings as well as the role of novel agents are discussed. First-line cisplatin-based polychemotherapy improves survival in the metastatic setting and is the standard of care. Approved regimens for subsequent-line therapy do not exist. Chemotherapy has a modest benefit in the neoadjuvant setting, but evidence is insufficient to justify its role in the adjuvant setting despite a possible benefit. Carboplatin cannot be substituted for cisplatin in fit patients, and the addition of taxane to a standard regimen cannot be recommended. Systemic chemotherapy plays a central role in the management of invasive bladder cancer in the metastatic and neoadjuvant settings, but its role in the adjuvant setting remains undefined. Neoadjuvant chemotherapy is underutilized and should be routinely used. Pathological downstaging strongly correlates with improved outcomes and may serve as a surrogate end point for survival. An urgent need exists for the development of novel therapeutic agents to improve outcomes.


Antibodies to pathogenic epitopes on type XVII collagen cause skin fragility in a complement-dependent and -independent manner.
In bullous pemphigoid (BP), the most prevalent autoimmune blistering disease, type XVII collagen (COL17) is targeted by circulating autoantibodies. BP is thought to be an autoantibody-mediated complement-fixing blistering disease, and a juxtamembranous noncollagenous 16A (NC16A) domain spanning Glu(490) to Arg(566) was proved to be the main pathogenic region on COL17, although precise pathogenic epitopes within NC16A have not been elucidated. In this study, we showed that injection of rabbit IgG Abs targeting Asp(522) to Gln(545) induced skin fragility associated with in vivo deposition of IgG and complement in neonatal COL17-humanized mice. Notably, immunoadsorption of rabbit anti-NC16A IgG Ab with this epitope (Asp(522) to Gln(545)) or the anti-NC16A IgG administered together with the peptides of this epitope as a decoy ameliorated skin fragility in the injected neonatal COL17-humanized mice compared with the anti-NC16A IgG alone even though all of the mice showed both IgG and complement deposition. These results led us to investigate an additional, complement-independent mechanism of skin fragility in the mice injected with anti-COL17 Abs. The rabbit anti-NC16A IgG depleted the expression of COL17 in cultured normal human keratinocytes, whereas immunoadsorption of the same IgG with this epitope significantly suppressed the depletion effect. Moreover, passive transfer of F(ab')(2) fragments of the human BP or rabbit IgG Abs against COL17 demonstrated skin fragility in neonatal COL17-humanized mice. In summary, this study reveals the importance of Abs directed against distinct epitopes on COL17, which induce skin fragility in complement-dependent as well as complement-independent ways.


Spatially resolved measurement of singlet delta oxygen by radar resonance-enhanced multiphoton ionization.
Coherent microwave Rayleigh scattering (Radar) from resonance-enhanced multiphoton ionization (REMPI) was demonstrated to directly and nonintrusively measure singlet delta oxygen, O(2)(a(1)Δ(g)), with high spatial resolution. Two different approaches, photodissociation of ozone and microwave discharge plasma in an argon and oxygen flow, were utilized for O(2)(a(1)Δ(g)) generation. The d(1)Π(g)←a(1)Δ(g) (3-0) and d(1)Π(g)←a(1)Δ(g) (1-0) bands of O(2)(a(1)Δ(g)) were detected by Radar REMPI for two different flow conditions. Quantitative absorption measurements using sensitive off-axis integrated cavity output spectroscopy (ICOS) was used simultaneously to evaluate the accuracy and sensitivity of the Radar REMPI technique. The detection limit of Radar REMPI was found to be comparable to the ICOS technique with a detection threshold of approximately 10(14) molecules/cm(3) but with a spatial resolution that was 8 orders of magnitude smaller than the ICOS technique.


Time course of histological progression in primary biliary cirrhosis.
Histological staging is used for stratification and assessment of treatment efficacy in therapeutic trials for primary biliary cirrhosis (PBC). Knowledge of the rate of progression of the histological changes would be helpful in the design (duration) and conduct of clinical trials. The histological stages were recorded for liver biopsies performed annually on 222 patients during a randomized, placebo-controlled clinical trial in which therapy with D-penicillamine (DPCA) was shown to be ineffective. These data were analyzed using a Markov model to describe the time course of histological progression in PBC. At study entry, 15 patients were stage I, 56 were stage II, 96 were stage III, and 55 were stage IV. Histological progression was observed after 1 year in 41%, 43%, and 35% of the patients, and after 2 years in 62%, 62%, and 50% of the patients who were stage I, stage II, and stage III at entry, respectively. After 4 years biopsies showed cirrhosis in 31% and 50% of the patients in stage I and stage II at entry, respectively. A minority (20%) of the precirrhotic patients showed histological stability; sustained histological regression was rarely observed (2%). Our data suggest that a majority of patients with PBC will progress histologically within 2 years. The distribution of histological stages over time may be helpful in determining the number of patients and length of time necessary to appreciate a treatment effect on histological progression in clinical trials for PBC.


Prostatic Abscess due to Burkholderia pseudomallei: Facilitating Diagnosis to Optimize Management.
Prostatic involvement is common in men with melioidosis. Indeed, some clinicians recommend radiological screening of all male patients as an undrained prostatic abscess may result in relapse of this potentially fatal disease. However, sophisticated medical imaging is frequently unavailable in the remote and resource-poor locations where patients are managed. In this retrospective study from Queensland, Australia, 22/144 (15%) men with melioidosis had a radiologically confirmed prostatic abscess. The absence of urinary symptoms had a negative predictive value (NPV) (95% confidence interval [CI]) for prostatic abscess of 96% (90-99%), whereas a urinary leukocyte count of < 50 × 106/L had an NPV (95% CI) of 100% (94-100%). A simple clinical history and basic laboratory investigations appear to exclude significant prostatic involvement relatively reliably and might be used to identify patients in whom radiological evaluation of the prostate is unnecessary. This may be particularly helpful in locations where radiological support is limited.


Rice Defense Mechanisms Against Cochliobolus miyabeanus and Magnaporthe grisea Are Distinct.
ABSTRACT Responses of rice to Magnaporthe grisea and Cochliobolus miyabeanus were compared. In Tetep, a rice cultivar resistant to both fungi, pathogen inoculation rapidly triggered the hypersensitive reaction (HR), resulting in microscopic cell death. In rice cv. Nakdong, susceptible to both pathogens, M. grisea did not cause HR, whereas C. miyabeanus caused rapid cell death similar to that associated with HR, which appeared similar to that observed in cv. Tetep, yet failed to block fungal ramification. Treatment with conidial germination fluid (CGF) from C. miyabeanus induced rapid cell death in both cultivars, suggesting the presence of phytotoxins in CGF. Pretreatment of cv. Nakdong with CGF significantly increased resistance to M. grisea, while the same treatment was ineffective against C. miyabeanus. Similarly, in cv. Nakdong, benzothiadiazole (BTH) significantly increased resistance to M. grisea, but was ineffective against C. miyabeanus. Methyl jasmonate (MeJA) treatment appeared to be ineffective against either fungus. Increased resistance of cv. Nakdong to M. grisea by BTH or CCF treatment was correlated with more rapid induction of three monitored PR genes. Application of MeJA resulted in the expression of JAmyb in cv. Nakdong being induced faster than in untreated plants in response to M. grisea infection. In contrast, the expression pattern of the PR and JAmyb genes in response to C. miyabeanus was nearly identical between cvs. Nakdong and Tetep, and neither BTH nor MeJA treatment significantly modified their expression patterns in response to C. miyabeanus infection. Our results suggest that rice employs distinct mechanisms for its defense against M. grisea versus C. miyabeanus.


Primary amebic meningoencephalomyelitis caused by Naegleria fowleri in a south-central black rhinoceros (Diceros bicornis minor).
CASE DESCRIPTION A 20-year-old female south-central black rhinoceros (Diceros bicornis minor) was evaluated because of an acute onset of CNS deficits. CLINICAL FINDINGS The rhinoceros had no history of illness. Clinical signs included acute lethargy, ataxia, and decreased appetite. Hematologic abnormalities included leukocytosis with neutrophilia and a profound left shift. Results of serum biochemical analysis revealed hypophosphatemia but no other abnormalities. Results of a quantitative PCR assay for West Nile virus and an assay for anti-Neosporum caninum antibodies in serum were negative; the patient was seropositive for multiple Leptospira serovars. TREATMENT AND OUTCOME Antimicrobials and anti-inflammatory agents were administered, but the condition of the rhinoceros worsened overnight; despite treatment with additional anti-inflammatory and antimicrobial agents, IV fluids, and thiamine, it became obtunded and died of respiratory arrest ≤ 24 hours later. Necropsy revealed severe, diffuse, suppurative, and histiocytic meningo-encephalomyelitis involving the cerebrum, cerebellum, and spinal cord. Amebic trophozoites were observed on histologic examination of affected tissue. Infection with Naegleria fowleri was confirmed by results of immuno-histochemical analysis and a multiplex real-time PCR assay. CLINICAL RELEVANCE Findings suggested that south-central black rhinoceros are susceptible to the free-living ameba N fowleri. Ameba-induced meningoencephalomyelitis should be considered as a differential diagnosis for rhinoceros that have an acute onset of neurologic signs. Diagnosis of N fowleri infection in an animal has a profound public health impact because of potential human exposure from the environment and the high fatality rate in people with N fowleri infection.


Effects of Exercise on Cognitive Performance in Children and Adolescents with ADHD: Potential Mechanisms and Evidence-based Recommendations.
Attention Deficit Hyperactivity Disorder (ADHD) is a neurodevelopmental disorder with a complex symptomatology, and core symptoms as well as functional impairment often persist into adulthood. Recent investigations estimate the worldwide prevalence of ADHD in children and adolescents to be ~7%, which is a substantial increase compared to a decade ago. Conventional treatment most often includes pharmacotherapy with central nervous stimulants, but the number of non-responders and adverse effects call for treatment alternatives. Exercise has been suggested as a safe and low-cost adjunctive therapy for ADHD and is reported to be accompanied by positive effects on several aspects of cognitive functions in the general child population. Here we review existing evidence that exercise affects cognitive functions in children with and without ADHD and present likely neurophysiological mechanisms of action. We find well-described associations between physical activity and ADHD, as well as causal evidence in the form of small to moderate beneficial effects following acute aerobic exercise on executive functions in children with ADHD. Despite large heterogeneity, meta-analyses find small positive effects of exercise in population-based control (PBC) children, and our extracted effect sizes from long-term interventions suggest consistent positive effects in children and adolescents with ADHD. Paucity of studies probing the effect of different exercise parameters impedes finite conclusions in this regard. Large-scale clinical trials with appropriately timed exercise are needed. In summary, the existing preliminary evidence suggests that exercise can improve cognitive performance intimately linked to ADHD presentations in children with and without an ADHD diagnosis. Based on the findings from both PBC and ADHD children, we cautiously provide recommendations for parameters of exercise.


Localization of organic anion transporting polypeptide 4 (Oatp4) in rat liver and comparison of its substrate specificity with Oatp1, Oatp2 and Oatp3.
Organic anion transporting polypeptides (rodents: Oatps; human: OATPs) are involved in the absorption and elimination of a wide variety of structurally unrelated amphipathic organic compounds. Several members of this protein family mediate the uptake of substrates across the basolateral membrane of hepatocytes as the first step in hepatic elimination. In contrast to the well-characterized Oatp1 and Oatp2, the localization and substrate specificity of the recently cloned Oatp4 have not been investigated in detail. Therefore, we raised an antibody against the C-terminal end of Oatp4 and localized this 85-kDa protein to the basolateral membrane of rat hepatocytes. Similar to Oatp1 and Oatp2, Oatp4 is a multispecific transporter with high affinities for bromosulfophthalein, dehydroepiandrosterone sulfate, leukotriene C4, and anionic peptides. In addition, we compared the substrate specificity of Oatp4 to that of Oatp3, which so far has mainly been shown to mediate intestinal bile acid transport. Oatp3 had a similar broad substrate specificity, but in general much lower affinities than Oatp4. Thus, while Oatp4 seems to work in concert with Oatp1 and Oatp2 in the basolateral membrane of rat hepatocytes, Oatp3 is a multispecific transport system in the small intestine.


Extract of Ginkgo biloba EGb 761 facilitates fear conditioning measured by fear-potentiated startle.
Extract of Ginkgo biloba EGb 761 has been used in the treatment of various common geriatric complaints including vertigo, short-term memory loss, hearing loss, lack of attention, vigilance and cerebral vascular disorder. Recent results suggest that it can serve as a cognitive enhancer and anti-stress buffer. It raises a possibility that EGb 761 may be involved in the fear conditioning. In this study, we used fear-potentiated startle (FPS) to evaluate the possible effects of EGb 761 on the acquisition stage of fear conditioning. Our results showed that administration of EGb 761 30 min prior to the conditioning facilitated acquisition of conditioned fear in a dose dependent manner. No significant differences had been observed in either basal startle response or shock activity. These results indicated that the facilitation effect of EGb 761 was not the result of impaired basal startle response or enhanced pain perception. Subsequent control experiment results indicated that the facilitation effect of EGb 761 on the acquisition was not due to anxiogenic effect or non-specific effect. Our data present the first evidence that EGb 761 can enhance fear memory formation rather than serve as an anti-stress buffer.


Collagen XV inhibits epithelial to mesenchymal transition in pancreatic adenocarcinoma cells.
Collagen XV (COLXV) is a secreted non-fibrillar collagen found within basement membrane (BM) zones of the extracellular matrix (ECM). Its ability to alter cellular growth in vitro and to reduce tumor burden and increase survival in vivo support a role as a tumor suppressor. Loss of COLXV during the progression of several aggressive cancers precedes basement membrane invasion and metastasis. The resultant lack of COLXV subjacent to the basement membrane and subsequent loss of its interactions with other proteins in this zone may directly impact tumor progression. Here we show that COLXV significantly reduces invasion of pancreatic adenocarcinoma cells through a collagen I (COLI) matrix. Moreover, we demonstrate that epithelial to mesenchymal transition (EMT) in these cells, which is recapitulated in vitro by cell scattering on a COLI substrate, is inhibited by over-expression of COLXV. We identify critical collagen-binding surface receptors on the tumor cells, including the discoidin domain receptor 1 (DDR1) and E-Cadherin (E-Cad), which interact with COLXV and appear to mediate its function. In the presence of COLXV, the intracellular redistribution of E-Cad from the cell periphery, which is associated with COLI-activated EMT, is inhibited and concurrently, DDR1 signaling is suppressed. Furthermore, continuous exposure of the pancreatic adenocarcinoma cells to high levels of COLXV suppresses endogenous levels of N-Cadherin (N-Cad). These data reveal a novel mechanism whereby COLXV can function as a tumor suppressor in the basement membrane zone.


The effect of arm position on the dosimetry of thoracic stereotactic ablative radiation therapy using volumetric modulated arc therapy.
Patient comfort and positioning stability may be improved in the arms down (AD) compared with the typical arms up (AU) position in thoracic stereotactic ablative radiation therapy (SABR). We compared plan quality for AD vs AU when using volumetric modulated arc therapy (VMAT), and evaluated the sensitivity of AD plans to arm positioning variability. We took plans of 14 patients with 17 lung tumors treated with thoracic SABR using VMAT in the AD position and simulated the same treatments in the AU position by re-optimizing after digitally removing the ipsilateral arm. To evaluate the sensitivity of AD plans to arm positioning variability, all plans were recalculated without re-optimization after assigning water density to the ipsilateral arm (AD-W) and then digitally shifting the arm 2.5 cm anterolaterally (AD-WS). Between AD and AU plans, statistically significant but clinically insignificant (all original planning constraints met) differences were found for the following parameters: mean planning target volume maximum dose, difference of 2.3% of prescription dose (P = .049); mean intermediate dose conformity index, difference of 0.27 (P = .012); median percent lung volume receiving a minimum of 10, 20, and 30 Gy (V10, V20, and V30), differences of 0.5%, 0.2%, and 0.1%, respectively (P = .040, .007, and .001); and median spinal cord maximum dose, difference of 33.5 cGy (P = .017). Similarly, between AD-W and AD-WS plans, statistically significant but clinically insignificant differences were found for median lung V20 and V30, difference of 0.0% for both (P = .034 and .016, by matched pair analysis). Our exploratory planning study suggests that when using VMAT for lung tumor SABR, AD and AU positioning achieve clinically equivalent plan quality, and AD plans are insensitive to relatively large variability in arm position.


Effects of Fabric Counts and Weave Designs on the Properties of Laminated Woven Kenaf/Carbon Fibre Reinforced Epoxy Hybrid Composites.
The effects of different fabric materials namely weave designs (plain and satin) and fabric counts (5 × 5 and 6 × 6) on the properties of laminated woven kenaf/carbon fibre reinforced epoxy hybrid composites were evaluated. The hybrid composites were fabricated from two types of fabric, i.e., woven kenaf that was made from a yarn of 500tex and carbon fibre, by using vacuum infusion technique and epoxy resin as matrix. The panels were tested for tensile, flexural, and impact strengths. The results have revealed that plain fabric is more suitable than satin fabric for obtaining high tensile and impact strengths. Using a fabric count of 5 × 5 has generated composites that are significantly higher in flexural modulus as compared to 6 × 6 which may be attributed to their structure and design. The scanned electron micrographs of the fractured surfaces of the composites demonstrated that plain woven fabric composites had better adhesion properties than satin woven fabric composites, as indicated by the presence of notably lower amount of fibre pull out.


Outcome evaluations in gout.
Methods to measure outcomes in gout still require consensus and validation. This Special Interest Group was assembled to identify domains of interest and is now evaluating a series of outcomes for features of acute gouty arthritis and chronic gout. To accomplish this, working groups have been formed and domains identified. Delphi methodology has been used to address gouty flares as an outcome of greatest interest. Studies addressing other outcome measures were reported at the OMERACT 8 meeting and validation has begun on some outcomes. There has been progress on developing a definition of a flare, and validating reproducibility of some chronic gout outcome measures in some domains, such as tophus size and patient perceptions. Use of these outcomes as well as a health-related quality of life measure are being studied in clinical trials. Pain on a Likert scale appears to be a valid outcome in acute gout. Final validation of these outcomes has not yet been achieved. In summary, the unique problems of evaluating outcomes in gout are finally being addressed. While no measures are available for use yet, an agenda has been developed.


Transforming growth factor-beta 1 and -beta 2 positively regulate TGF-beta 1 mRNA expression in trabecular cells.
To determine whether transforming growth factor (TGF)-beta 1 and -beta 2 and basic fibroblast growth factor (bFGF) induce the gene expression of TGF-beta 1 in the first-passage trabecular meshwork cells of the eye. Trabecular meshwork cells were cultured from fresh porcine eyes and treated with 1 ng/ml of TGF-beta 1, TGF-beta 2, or bFGF for 1 hour. Cells maintained in serum-free medium were used as controls. Total cellular RNA was extracted, and the first-strand cDNA was synthesized. Multiplex polymerase chain reaction (PCR) and competitive PCR were performed on aliquots of the cDNAs by using either endogenous (glyceraldehyde-3-phosphate dehydrogenase [G3PDH]) or exogenous sequence (PCR mimic for TGF-beta 1) as internal standards, respectively. The obtained products were quantitated by laser densitometry, and statistical analysis was performed. The findings show that trabecular cells in vitro express the TGF-beta 1 messenger RNA constitutively. Both the techniques of multiplex PCR and competitive PCR demonstrated that the addition of either TGF-beta 1 or TGF-beta 2 at a concentration present in normal aqueous humor increased the mRNA levels of TGF-beta 1 by 2.82-to 3.07-fold over the controls, and these results were statistically significant (P < 0.01). Basic fibroblast growth factor did not have an effect on TGF-beta 1 expression (P < 0.05). Transforming growth factor-beta 1 activates its own gene expression in trabecular cells. Considering the multifunctional property of this cytokine, which includes increased deposition of extracellular matrix material and growth inhibition of trabecular cells, a change in its concentration within the eye would have a profound effect because of this autoinductive activity. Transforming growth factor-beta 2 treatment of trabecular cells also increased their expression of the TGF-beta 1 gene. The authors previously showed that the level of TGF-beta 2 in the aqueous humor of glaucomatous eyes is significantly higher than that of age-matched nonglaucomatous controls. The current finding suggests that this growth modulator may exert its effects directly on the trabecular cells or that it may act indirectly through upregulating the production of TGF-beta 1.


Three-dimensional quasi-periodic shifted Green function throughout the spectrum, including Wood anomalies.
This work, part II in a series, presents an efficient method for evaluation of wave scattering by doubly periodic diffraction gratings at or near what are commonly called 'Wood anomaly frequencies'. At these frequencies, there is a grazing Rayleigh wave, and the quasi-periodic Green function ceases to exist. We present a modification of the Green function by adding two types of terms to its lattice sum. The first type are transversely shifted Green functions with coefficients that annihilate the growth in the original lattice sum and yield algebraic convergence. The second type are quasi-periodic plane wave solutions of the Helmholtz equation which reinstate certain necessary grazing modes without leading to blow-up at Wood anomalies. Using the new quasi-periodic Green function, we establish, for the first time, that the Dirichlet problem of scattering by a smooth doubly periodic scattering surface at a Wood frequency is uniquely solvable. We also present an efficient high-order numerical method based on this new Green function for scattering by doubly periodic surfaces at and around Wood frequencies. We believe this is the first solver able to handle Wood frequencies for doubly periodic scattering problems in three dimensions. We demonstrate the method by applying it to acoustic scattering.


Whole-genome comparisons of Penicillium spp. reveals secondary metabolic gene clusters and candidate genes associated with fungal aggressiveness during apple fruit decay.
Blue mold is a postharvest rot of pomaceous fruits caused by Penicillium expansum and a number of other Penicillium species. The genome of the highly aggressive P. expansum strain R19 was re-sequenced and analyzed together with the genome of the less aggressive P. solitum strain RS1. Whole genome scale similarities and differences were examined. A phylogenetic analysis of P. expansum, P. solitum, and several closely related Penicillium species revealed that the two pathogens isolated from decayed apple with blue mold symptoms are not each other's closest relatives. Among a total of 10,560 and 10,672 protein coding sequences respectively, a comparative genomics analysis revealed 41 genes in P. expansum R19 and 43 genes in P. solitum RS1 that are unique to these two species. These genes may be associated with pome fruit-fungal interactions, subsequent decay processes, and mycotoxin accumulation. An intact patulin gene cluster consisting of 15 biosynthetic genes was identified in the patulin producing P. expansum strain R19, while only a remnant, seven-gene cluster was identified in the patulin-deficient P. solitum strain. However, P. solitum contained a large number of additional secondary metabolite gene clusters, indicating that this species has the potential capacity to produce an array of known as well as not-yet-identified products of possible toxicological or biotechnological interest.


A retrospective analysis of self-reported symptoms from 761 consecutive new patients presenting to a Neuro Emotional Technique chiropractic clinic.
To describe the profile of patients presenting to a private chiropractic clinic specialising in Neuro Emotional Technique (NET) and to identify trends in the presentation of symptoms from these patients. 761 consecutive new patients presented to a large, multi-doctor chiropractic clinic in which practitioners all adopt a similar philosophical paradigm and practice NET From January 2005 to December 2005, self-referred patients completed a new patient questionnaire, in which they self-reported one primary complaint for why they were visiting the practitioner. Predetermined patient information was entered manually into a database and basic descriptive statistics extracted. 67.3% of participants were female and 32.6% of the participants were between the ages of 31 and 40. 54.8% of patients presented with a primary musculoskeletal complaint and 36.0% a non-musculoskeletal complaint. Of the musculoskeletal complaints, 40.8% of patients presented with back pain, 20.9% with neck pain and 11.5% with shoulder pain. The most common form of non-musculoskeletal complaint was immune and recurrent infections (13.9%), stress and anxiety (12.8%) and depression (10.9%). 41.4% of participants reported a first time complaint, however, of the patients who had had the presenting complaint before 60.7% reported as having the complaint for greater than 1 year. Musculoskeletal and non-musculoskeletal participants had similar pain profiles. This retrospective analysis is the first comprehensive description of the scope of NET patients and their presenting complaints. The patient profile of this NET clinic has a higher degree of non-musculoskeletal patients than that usually reported in non-NET chiropractic offices, and other forms of chiropractic previously described in the literature. Further cross sectional research is required to determine if this particular clinic is indicative of all NET practices and whether the presenting symptoms, especially the non-musculoskeletal, are resolved with NET.


Management and prevention of mycotoxins in peanuts.
Contamination of peanuts with mycotoxins, particularly aflatoxins, is a worldwide problem that affects both food safety and agricultural economies. Most countries have adopted regulations that limit the quantity of aflatoxins in food and feed to 20 microg kg(-1) or less; however, environmental conditions in most of the world where peanuts are produced and stored often make it difficult or impossible to attain such low concentrations. In addition to aflatoxins, peanuts are often contaminated with cyclopiazonic acid (CPA). Both mycotoxins are produced by Aspergillus flavus, a ubiquitous fungus that can infect and grow in peanuts under both pre- and post-harvest conditions. Management of mycotoxin contamination in peanuts generally involves removal of high-risk components from shelled lots or the removal of individual, highly contaminated nuts. This is accomplished by various processes such as screening, kernel sizing, electronic colour sorting, hand sorting, and blanching followed by electronic colour sorting. Recently, biological control technology has been developed that prevents much of the contamination that might otherwise occur. Biocontrol is based on competitive exclusion whereby a dominant population of a non-toxigenic strain of A. flavus is established in the soil before peanuts are subjected to conditions favouring contamination. The applied strain competes with toxigenic strains for infection sites, resulting in significantly reduced concentrations of aflatoxins in peanuts. Monitoring of the first commercial use of the technology showed that aflatoxins were reduced by an average of 85% in farmers' stock peanuts and by as much as 98% in shelled, edible grade peanuts.


Maintenance bevacizumab-pemetrexed after first-line cisplatin-pemetrexed-bevacizumab for advanced nonsquamous nonsmall-cell lung cancer: updated survival analysis of the AVAPERL (MO22089) randomized phase III trial.
The randomized, phase III AVAPERL trial evaluated the safety and efficacy of bevacizumab maintenance with or without pemetrexed in nonsquamous nonsmall-cell lung cancer (nsNSCLC). Progression-free survival (PFS) was significantly prolonged with bevacizumab-pemetrexed, but overall survival (OS) data were immature. In this article, we report an independent, updated analysis of survival outcomes in AVAPERL. Patients with advanced nsNSCLC received first-line bevacizumab (7.5 mg/kg), cisplatin (75 mg/m(2)), and pemetrexed (500 mg/m(2)) every 3 weeks (q3w) for four cycles. Nonprogressing patients were randomized to maintenance bevacizumab (7.5 mg/kg) or bevacizumab-pemetrexed (500 mg/m(2)) q3w until progression or consent withdrawal. The primary end point of the trial was PFS; in this independent OS analysis, participating study centers were contacted to collect survival data on patients still alive at the time of the first analysis. A total of 376 patients received induction treatment. Disease control was confirmed in 71.9% of patients; 253 patients were randomized to maintenance treatment with bevacizumab (n = 125) or bevacizumab-pemetrexed (n = 128). At a median follow-up of 14.8 months, patients allocated to bevacizumab-pemetrexed had significantly improved PFS versus those on bevacizumab when measured from randomization [7.4 versus 3.7 months, hazard ratio (HR), 0.57, 95% confidence interval (CI) 0.44-0.75); P < 0.0001]. OS events occurred in 58% of all patients. OS was numerically longer with bevacizumab-pemetrexed versus bevacizumab when measured from randomization [17.1 versus 13.2 months, HR 0.87 (0.63-1.21); P = 0.29]. Second-line therapy was administered in 77% and 70% of patients in the bevacizumab and bevacizumab-pemetrexed arms, respectively. No new adverse events were reported during this updated analysis. In an unselected population of nsNSCLC patients achieving disease control on platinum-based induction therapy, maintenance with bevacizumab-pemetrexed was associated with a nonsignificant increase in OS over bevacizumab alone.


Human Papillomavirus Vaccination and Age at First Sexual Activity, National Health and Nutrition Examination Survey.
The National Health and Nutrition Examination Survey (NHANES) collects information on human papillomavirus (HPV) vaccination history as well as sexual activity. We evaluated data from NHANES to assess report of HPV vaccination with ≥1 dose and 3 doses among females and males aged 11 to 26 years during 2007-2014. We also examined age at first vaccine dose and age at first sexual activity among females aged 14 to 26 years. Vaccination significantly increased in females aged 13 to 26 years, but not among 11- to 12-year-old girls, and remained low for both females and males. In NHANES 2011-2014, among females with known age at first vaccine dose, 43.1% reported having had sex before or in the same year they received their first HPV vaccine, and this varied by race/ethnicity. Clinicians should provide strong recommendations consistent with guidelines, including routine vaccination of girls and boys at age 11 or 12 years.


Regional cerebral blood flow alterations in unipolar depression.
Forty-seven symptomatic inpatients and outpatients with major depression (13 nonendogenous, 23 endogenous, and 11 psychotic by Research Diagnostic Criteria) were compared with 138 normal control subjects. Absolute regional cerebral blood flow (rCBF, ml/minute/100 g) was measured with 133Xe single photon emission computed tomography. Flow ratios (region of interest/global flow) and residual scores (the difference between patient flow ratios and expected normal flow ratios, as derived from the control population) were also computed. Results revealed significant age x region x depression subtype interactions for absolute, ratio, and residual flow data. Consequently, a test of group means (or analysis of covariance) could not be used to examine between-group differences. Multiple regression analyses were employed to study the effects of age on rCBF. This analysis revealed that different, though sometimes overlapping, regions exhibited different age effects on rCBF in different depressive subtypes. Thus, diagnostic-subtype-dependent age effects on rCBF precluded comparisons of mean values within or across regions for subject groups, but distinguished between symptomatic depressed patients and control subjects and among patient groups. Possible causes of such effects include variations in duration of illness or medication history or sensitization phenomena.


An open source based high content screening method for cell biology laboratories investigating cell spreading and adhesion.
Adhesion dependent mechanisms are increasingly recognized to be important for a wide range of biological processes, diseases and therapeutics. This has led to a rising demand of pharmaceutical modulators. However, most currently available adhesion assays are time consuming and/or lack sensitivity and reproducibility or depend on specialized and expensive equipment often only available at screening facilities. Thus, rapid and economical high-content screening approaches are urgently needed. We established a fully open source high-content screening method for identifying modulators of adhesion. We successfully used this method to detect small molecules that are able to influence cell adhesion and cell spreading of Swiss-3T3 fibroblasts in general and/or specifically counteract Nogo-A-Δ20-induced inhibition of adhesion and cell spreading. The tricyclic anti-depressant clomipramine hydrochloride was shown to not only inhibit Nogo-A-Δ20-induced cell spreading inhibition in 3T3 fibroblasts but also to promote growth and counteract neurite outgrowth inhibition in highly purified primary neurons isolated from rat cerebellum. We have developed and validated a high content screening approach that can be used in any ordinarily equipped cell biology laboratory employing exclusively freely available open-source software in order to find novel modulators of adhesion and cell spreading. The versatility and adjustability of the whole screening method will enable not only centers specialized in high-throughput screens but most importantly also labs not routinely employing screens in their daily work routine to investigate the effects of a wide range of different compounds or siRNAs on adhesion and adhesion-modulating molecules.


A unified model for the G1/S cell cycle transition.
Efficient S phase entry is essential for development, tissue repair, and immune defences. However, hyperactive or expedited S phase entry causes replication stress, DNA damage and oncogenesis, highlighting the need for strict regulation. Recent paradigm shifts and conflicting reports demonstrate the requirement for a discussion of the G1/S transition literature. Here, we review the recent studies, and propose a unified model for the S phase entry decision. In this model, competition between mitogen and DNA damage signalling over the course of the mother cell cycle constitutes the predominant control mechanism for S phase entry of daughter cells. Mitogens and DNA damage have distinct sensing periods, giving rise to three Commitment Points for S phase entry (CP1-3). S phase entry is mitogen-independent in the daughter G1 phase, but remains sensitive to DNA damage, such as single strand breaks, the most frequently-occurring lesions that uniquely threaten DNA replication. To control CP1-3, dedicated hubs integrate the antagonistic mitogenic and DNA damage signals, regulating the stoichiometric cyclin: CDK inhibitor ratio for ultrasensitive control of CDK4/6 and CDK2. This unified model for the G1/S cell cycle transition combines the findings of decades of study, and provides an updated foundation for cell cycle research.


Skin-related Quality of Life among Midwestern US Community-based Women with Breast Cancer Experiencing Radiodermatitis.
Little is known about the skin-related quality of life (QOL) among women receiving external radiotherapy (EBT) and who experience breast radiodermatitis. This pilot study aimed to describe the thoughts and experiences of women experiencing breast radiodermatitis of the breast at a comprehensive community cancer program. A printed survey was used to solicit feedback on the Dermatology Life Quality Index (DLQI) during the 5th week of EBT. An open-ended question inquired which DLQI-related issue was most important and why. A directed qualitative content analysis was conducted on the narrative responses. Twenty-eight women provided a response to the "most important" question. Sixty narratives led to the identification of 35 codes and six themes during content analysis. Themes included perspectives on having radiodermatitis, sensations caused by radiodermatitis, knowledge, and preparation for radiotherapy, prevention of radiodermatitis, emotions induced by skin changes, and physical appearance of the breast skin. The study results provide a glimpse into the perceptions of skin-related QOL among community-dwelling women who experienced breast radiodermatitis. Some women expressed that radiodermatitis had a profound impact on their QOL while other were surprised that EBT was easy compared to chemotherapy. Our findings parallel those found in a previous study conducted in an urban setting. Results provide insight into the thoughts and needs of women undergoing breast EBT. Assessing individual differences in skin-related QOL can provide needed information for tailoring care to the unique needs of each woman. Additional studies focusing specifically on skin-related QOL are needed.


Assessing nursing students' basic communication and interviewing skills: the development and testing of a rating scale.
This study explores the communication skills of a group of nursing students who were required to interview a simulated client as part of their studies. In order to assess the students and to improve the process of learning discrete skills, an instrument was developed and tested as part of this process. The subjects were 212 nurses enrolled in a bachelor of nursing programme, in New South Wales, Australia, who were studying a problem-based learning package the focus of which was 'alcohol early intervention'. The sub-groups within the sample included registered nurses, a significant percentage of whom had completed their basic nursing education in overseas countries. The Simulated Client Interview Rating Scale (SCIRS) was developed to assess basic humanistic communication skills as well as beginning motivational interviewing skills. The students were required to interview a simulated client and demonstrate competence in interviewing. This was assessed by the SCIRS which was completed by the students and the simulated clients. The instrument proved to be a reliable and valid means of assessing student interview technique as well as a flexible educational tool, while valuable insights into students' interviewing techniques were gained.


How children affect the mother/victim's process in intimate partner violence.
Witnessing intimate partner violence (IPV) causes physical and mental health problems for children. Children are one of multiple factors that a victim weighs as she manages the abusive relationship. Little has been written about how children affect the mother's decisions about the abuse or what assistance a mother wants from the children's physician in creating a nonabusive home. To consider the role children play in their mothers' management of abusive partners. Thirty-two mothers living in midwestern IPV shelters or participating in support groups were interviewed about their abuse stories, perceptions about the effects of the abuse on their children, and desires about IPV management in the health care setting. The interviews were audiotaped, transcribed, and analyzed by a team of researchers using thematic analysis. Children were an integral factor in the mothers' management of their abusive relationships. For more than half of the participants, something the children did or said catalyzed their seeking help. For some, the children's attachment to the abuser was a reason to delay seeking assistance. Based on these findings, we explored what mothers wanted from their children's physicians regarding their abusive relationships. Mothers talked about the delicate balance between education and blame, between offering help and becoming too intrusive, and between wanting the best for their children and fearing the involvement of child protective services. Children play an important role in mothers' management of their abusive relationships. From their children's physicians, participants wanted IPV screening and IPV resources. Some wanted the physician to educate them about how the IPV affected the children in a nonblaming manner.


Standards for the management of sickle cell disease in children.
Sickle cell disease is now the commonest genetic condition in England with a birth prevalence of 1 in 2000. It causes significant morbidity and mortality particularly in the early years unless prophylactic measures have been put in place. By the end of 2006 newborn screening had been introduced all over England, its prime aim being to minimise mortality from pneumococcal sepsis in the first few years of life. Plans are currently underway to roll out programmes in Scotland and Wales. Standards and guidelines for the care of children with sickle cell disease were written to accompany the newborn screening programme and to offer a management plan for those working in areas of low prevalence where resources and expertise were possibly less well developed. This article describes the existing evidence base for treatment and the current consensus of good practice. It is acknowledged that more work needs to be carried out to develop guidelines further.


Current strategies for monitoring men with localised prostate cancer lack a strong evidence base: observational longitudinal study.
The UK National Institute for Health and Clinical Excellence (NICE) guidance recommends conservative management of men with 'low-risk' localised prostate cancer, monitoring the disease using prostate-specific antigen (PSA) kinetics and re-biopsy. However, there is little evidence of the changes in PSA level that should alert to the need for clinical re-assessment. This study compares the alerts resulting from PSA kinetics and a novel longitudinal reference range approach, which incorporates age-related changes, during the monitoring of 408 men with localised prostate cancer. Men were monitored by regular PSA tests over a mean of 2.9 years, recording when a man's PSA doubling time fell below 2 years, PSA velocity exceeded 2 ng ml(-1) per year, or when his upper 10% reference range was exceeded. Prostate-specific antigen doubling time and PSA velocity alerted a high proportion of men initially but became unresponsive to changes with successive tests. Calculating doubling time using recent PSA measurements reduced the decline in response. The reference range method maintained responsiveness to changes in PSA level throughout the monitoring. The increasing unresponsiveness of PSA kinetics is a consequence of the underlying regression model. Novel methods are needed for evaluation in cohorts currently being managed by monitoring. Meanwhile, the NICE guidance should be cautious.


The development and evaluation of a geriatric emergency medicine curriculum. The SAEM Geriatric Emergency Medicine Task Force.
To summarize the processes used to develop a curriculum and model of care for the emergency medical treatment of elder patients and to assess the efficacy of the teaching material in a pilot course. A survey of emergency medicine (EM) residency directors and geriatric fellowship directors was used to identify key topics for inclusion in the didactic material. An interdisciplinary consensus process was used to develop didactic as well as teaching material in geriatric EM. Pretests and posttests were administered to 46 participants in the initial course to assess knowledge gain. Subjective course evaluations were also done. Test scores significantly increased from 54% correct on the pretest to 77% correct on the posttest (p < 0.001). Significant improvement in knowledge as judged by pretest and posttest results occurred in 6 of the 7 teaching modules. Subjective evaluations demonstrated good to excellent ratings for each module as well as the overall workshop. The process of developing a curriculum for geriatric EM is described. The initial training of instructors was effective in improving participants' knowledge of geriatric issues in EM. Participants considered the training to be effective. The effect of the training on the emergency care of elder persons remains to be determined.


Alpha lipoic acid possess dual antioxidant and lipid lowering properties in atherosclerotic-induced New Zealand White rabbit.
There is accumulating data demonstrated hypercholesterolemia and oxidative stress play an important role in the development of atherosclerosis. In the present study, a protective activity of alpha-lipoic acid; a metabolic antioxidant in hypercholesterolemic-induced animals was investigated. Eighteen adult male New Zealand White (NZW) rabbit were segregated into three groups labelled as group K, AT and ALA (n=6). While group K was fed with normal chow and acted as a control, the rest fed with 100 g/head/day with 1% high cholesterol diet to induce hypercholesterolemia. 4.2 mg/body weight of alpha lipoic acid was supplemented daily to the ALA group. Drinking water was given ad-libitum. The study was designed for 10 weeks. Blood sampling was taken from the ear lobe vein at the beginning of the study, week 5 and week 10 and plasma was prepared for lipid profile estimation and microsomal lipid peroxidation index indicated with malondialdehyde (MDA) formation. Animals were sacrificed at the end of the study and the aortas were excised for intimal lesion analysis. The results showed a significant reduction of lipid peroxidation index indicated with low MDA level (p<0.05) in ALA group compared to that of the AT group. The blood total cholesterol (TCHOL) and low density lipoprotein (LDL) levels were found to be significantly low in ALA group compared to that of the AT group (p<0.05). Histomorphometric intimal lesion analysis of the aorta showing less of atheromatous plaque formation in alpha lipoic acid supplemented group (p<0.05) compared to that of AT group. These findings suggested that apart from its antioxidant activity, alpha lipoic acid may also posses a lipid lowering effect indicated with low plasma TCHOL and LDL levels and reduced the athero-lesion formation in rabbits fed a high cholesterol diet.


Histone modification analysis by chromatin immunoprecipitation from a low number of cells on a microfluidic platform.
Histone modifications are important epigenetic mechanisms involved in eukaryotic gene regulation. Chromatin immunoprecipitation (ChIP) assay serves as the primary technique to characterize the genomic locations associated with histone modifications. However, traditional tube-based ChIP assays rely on large numbers of cells as well as laborious and time-consuming procedures. Here we demonstrate a novel microfluidics-based native ChIP assay which dramatically reduces the required cell number and the assay time by conducting cell collection, lysis, chromatin fragmentation, immunoprecipitation, and washing on a microchip. Coupled with real-time PCR, our assay permits the analysis of histone modifications from as few as ~50 cells within 8.5 h. We envision that our method will provide a new approach for the analysis of epigenetic regulations and protein-DNA interactions in general, based on scarce cell samples such as those derived from animals and patients.


An evaluation tool for Myofascial Adhesions in Patients after Breast Cancer (MAP-BC evaluation tool): Concurrent, face and content validity.
To investigate the concurrent, face and content validity of an evaluation tool for Myofascial Adhesions in Patients after Breast Cancer (MAP-BC evaluation tool). 1) Concurrent validity of the MAP-BC evaluation tool was investigated by exploring correlations (Spearman's rank Correlation Coefficient) between the subjective scores (0 -no adhesions to 3 -very strong adhesions) of the skin level using the MAP-BC evaluation tool and objective elasticity parameters (maximal skin extension and gross elasticity) generated by the Cutometer Dual MPA 580. Nine different examination points on and around the mastectomy scar were evaluated. 2) Face and content validity were explored by questioning therapists experienced with myofascial therapy in breast cancer patients about the comprehensibility and comprehensiveness of the MAP-BC evaluation tool. 1) Only three meaningful correlations were found on the mastectomy scar. For the most lateral examination point on the mastectomy scar a moderate negative correlation (-0.44, p = 0.01) with the maximal skin extension and a moderate positive correlation with the resistance versus ability of returning or 'gross elasticity' (0.42, p = 0.02) were found. For the middle point on the mastectomy scar an almost moderate positive correlation with gross elasticity was found as well (0.38, p = 0.04) 2) Content and face validity have been found to be good. Eighty-nine percent of the respondent found the instructions understandable and 98% found the scoring system obvious. Thirty-seven percent of the therapists suggested to add the possibility to evaluate additional anatomical locations in case of reconstructive and/or bilateral surgery. The MAP-BC evaluation tool for myofascial adhesions in breast cancer patients has good face and content validity. Evidence for good concurrent validity of the skin level was found only on the mastectomy scar itself.


Guillain-Barré Syndrome Following Zika Virus Infection Is Associated With a Diverse Spectrum of Peripheral Nerve Reactive Antibodies.
Recent outbreaks of Zika virus (ZIKV) in South and Central America have highlighted significant neurologic side effects. Concurrence with the inflammatory neuropathy Guillain-Barré syndrome (GBS) is observed in 1:4,000 ZIKV cases. Whether the neurologic symptoms of ZIKV infection are immune mediated is unclear. We used rodent and human live cellular models to screen for anti-peripheral nerve reactive IgG and IgM autoantibodies in the sera of patients with ZIKV with and without GBS. In this study, 52 patients with ZIKV-GBS were compared with 134 ZIKV-infected patients without GBS and 91 non-ZIKV controls. Positive sera were taken forward for target identification by immunoprecipitation and mass spectrometry, and candidate antigens were validated by ELISA and cell-based assays. Autoantibody reactions against glycolipid antigens were also screened on an array. Overall, IgG antibody reactivities to rat Schwann cells (SCs) (6.5%) and myelinated cocultures (9.6%) were significantly higher, albeit infrequent, in the ZIKV-GBS group compared with all controls. IgM antibody immunoreactivity to dorsal root ganglia neurones (32.3%) and SCs (19.4%) was more frequently observed in the ZIKV-GBS group compared with other controls, whereas IgM reactivity to cocultures was as common in ZIKV and non-ZIKV sera. Strong axonal-binding ZIKV-GBS serum IgG antibodies from 1 patient were confirmed to react with neurofascin 155 and 186. Serum from a ZIKV-infected patient without GBS displayed strong myelin-binding and putative antilipid antigen reaction characteristics. There was, however, no significant association of ZIKV-GBS with any known antiglycolipid antibodies. Autoantibody responses in ZIKV-GBS target heterogeneous peripheral nerve antigens suggesting heterogeneity of the humoral immune response despite a common prodromal infection.


A new weapon against an old target.
High expression of folate receptors is characteristic for the effector cell population of synovial macrophages in synovial inflammation. A new drug conjugate, EC0746, targets the folate inhibitor aminopterin to folate receptors. In vitro studies show that this conjugate acts antiproliferatively and inhibits cytokine production by macrophage cell lines. Moreover, it shows strong anti-arthritic effects in the rat model of adjuvant-induced arthritis in vivo. Toxicity of aminopterin was reduced 40-fold by using equimolar doses of the drug conjugate. In conclusion, this new treatment approach has the potential to further improve the most successful principle of folate inhibition in the treatment of arthritis.


Constitutive activation of G-proteins by polycystin-1 is antagonized by polycystin-2.
Polycystin-1 (PC1), a 4,303-amino acid integral membrane protein of unknown function, interacts with polycystin-2 (PC2), a 968-amino acid alpha-type channel subunit. Mutations in their respective genes cause autosomal dominant polycystic kidney disease. Using a novel heterologous expression system and Ca(2+) and K(+) channels as functional biosensors, we found that full-length PC1 functioned as a constitutive activator of G(i/o)-type but not G(q)-type G-proteins and modulated the activity of Ca(2+) and K(+) channels via the release of Gbetagamma subunits. PC1 lacking the N-terminal 1811 residues replicated the effects of full-length PC1. These effects were independent of regulators of G-protein signaling proteins and were lost in PC1 mutants lacking a putative G-protein binding site. Co-expression with full-length PC2, but not a C-terminal truncation mutant, abrogated the effects of PC1. Our data provide the first experimental evidence that full-length PC1 acts as an untraditional G-protein-coupled receptor, activity of which is physically regulated by PC2. Thus, our study strongly suggests that mutations in PC1 or PC2 that distort the polycystin complex would initiate abnormal G-protein signaling in autosomal dominant polycystic kidney disease.


Evaluating plant immunity using mass spectrometry-based metabolomics workflows.
Metabolic processes in plants are key components of physiological and biochemical disease resistance. Metabolomics, the analysis of a broad range of small molecule compounds in a biological system, has been used to provide a systems-wide overview of plant metabolism associated with defense responses. Plant immunity has been examined using multiple metabolomics workflows that vary in methods of detection, annotation, and interpretation, and the choice of workflow can significantly impact the conclusions inferred from a metabolomics investigation. The broad range of metabolites involved in plant defense often requires multiple chemical detection platforms and implementation of a non-targeted approach. A review of the current literature reveals a wide range of workflows that are currently used in plant metabolomics, and new methods for analyzing and reporting mass spectrometry (MS) data can improve the ability to translate investigative findings among different plant-pathogen systems.


Oxidative metabolism of omeprazole in human liver microsomes: cosegregation with S-mephenytoin 4'-hydroxylation.
Oxidative metabolism of omeprazole (OPZ) was studied in 14 human liver microsomes in relation to the 4'-hydroxylation capacity of S-mephenytoin. The formation of 5-hydroxyomeprazole and omeprazole sulfone (OPZ-SFN) from OPZ exhibited a biphasic kinetic behavior, indicating that at least two distinct enzymes are involved in either of the metabolic pathways of OPZ. By using a two-enzyme kinetic approach, the activities were described by high (Km1 and Vmax1)- and low-affinity components (Km2 and Vmax2). The respective mean (+/- S.D.) kinetic parameters for 5-hydroxylation and sulfoxidation were: Km1 = 6.0 +/- 2.4 and 10.2 +/- 7.2 microM and Vmax1 = 88.0 +/- 70.2 and 66.9 +/- 53.9 pmol/mg/min; Km2 = 106 +/- 127 and 482 +/- 472 microM and Vmax2 = 116 +/- 88 and 299 +/- 131 pmol/mg/min. Among these kinetic parameters, only the Vmax1 of 5-hydroxylation gave a close correlation with the corresponding parameter of S-mephenytoin (rs = 0.911, P < .01). In addition, OPZ and S-mephenytoin inhibited competitively each other's metabolism with the respective Ki values of 2.0 and 162 microM. Interestingly, OPZ-SFN also inhibited competitively 4'-hydroxylation of S-mephenytoin with a Ki value of 8.2 microM. Moreover, polyclonal antibodies raised against S-mephenytoin 4'-hydroxylase (P450 form) partially inhibited the 5-hydroxylation of OPZ, whereas no inhibition was observed for the sulfoxidation. These findings suggest that S-mephenytoin 4'-hydroxylase is an enzyme primarily responsible for the 5-hydroxylation of OPZ and further metabolism of OPZ-SFN, but not for the sulfoxidation of OPZ in human liver microsomes.(ABSTRACT TRUNCATED AT 250 WORDS)


Trade-offs and synergies between carbon storage and livelihood benefits from forest commons.
Forests provide multiple benefits at local to global scales. These include the global public good of carbon sequestration and local and national level contributions to livelihoods for more than half a billion users. Forest commons are a particularly important class of forests generating these multiple benefits. Institutional arrangements to govern forest commons are believed to substantially influence carbon storage and livelihood contributions, especially when they incorporate local knowledge and decentralized decision making. However, hypothesized relationships between institutional factors and multiple benefits have never been tested on data from multiple countries. By using original data on 80 forest commons in 10 countries across Asia, Africa, and Latin America, we show that larger forest size and greater rule-making autonomy at the local level are associated with high carbon storage and livelihood benefits; differences in ownership of forest commons are associated with trade-offs between livelihood benefits and carbon storage. We argue that local communities restrict their consumption of forest products when they own forest commons, thereby increasing carbon storage. In showing rule-making autonomy and ownership as distinct and important institutional influences on forest outcomes, our results are directly relevant to international climate change mitigation initiatives such as Reduced Emissions from Deforestation and Forest Degradation (REDD) and avoided deforestation. Transfer of ownership over larger forest commons patches to local communities, coupled with payments for improved carbon storage can contribute to climate change mitigation without adversely affecting local livelihoods.


Evidence of Change of Intention in Picking Situations.
Intending to perform an action and then immediately executing it is a mundane process. The cognitive and neural mechanisms involved in this process of "proximal" intention formation and execution, in the face of multiple options to choose from, are not clear, however. Especially, it is not clear how intentions are formed when the choice makes no difference. Here we used behavioral and electrophysiological measures to investigate the temporal dynamics of proximal intention formation and "change of intention" in a free picking scenario, in which the alternatives are on a par for the participant. Participants pressed a right or left button following either an instructive visible arrow cue or a visible neutral "free-choice" cue, both preceded by a masked arrow prime. The goal of the prime was to induce a bias toward pressing the left or right button. Presumably, when the choice is arbitrary, such bias should determine the decision. EEG lateralized readiness potentials and EMG measurements revealed that the prime indeed induced an intention to move in one direction. However, we discovered a signature of "change of intention" in both the Instructed and Free-choice decisions. These results suggest that, even in arbitrary choices, biases present in the neural system for choosing one or another option may be overruled and point to a curious "picking deliberation" phenomenon. We discuss a possible neural scenario that could explain this phenomenon.


Positive and cost-effectiveness effect of spa therapy on the resumption of occupational and non-occupational activities in women in breast cancer remission: a French multicentre randomised controlled trial.
The main aim was to assess the effects of a spa treatment on the resumption of occupational and non-occupational activities and the abilities of women in breast cancer remission. A cost-effectiveness analysis (CEA) was also performed. A multicentre randomised controlled trial was carried out between 2008 and 2010 in the University Hospital of Auvergne and two private hospitals in Clermont-Ferrand, France. Eligible patients were women in complete breast cancer remission without contraindication for physical activities or cognitive disorders and a body mass index between 18.5 and 40 kg/m(2). The intervention group underwent spa treatment combined with consultation with dietician whereas the control underwent consultations with the dietician only. Of the 181 patients randomised, 92 and 89 were included in the intervention and the control groups, respectively. The CEA involved 90 patients, 42 from the intervention group and 48 from the control group. The main results showed a higher rate of resumption of occupational activities in the intervention group (p = 0.0025) and a positive effect of the intervention on the women's ability to perform occupational activities 12 months after the beginning of the study (p = 0.0014), and on their ability to perform family activities (p = 0.033). The stay in a thermal centre was cost-effective at 12 months. Spa treatment is a cost-effective strategy to improve resumption of occupational and non-occupational activities and the abilities of women in breast cancer remission.


Proteomic Differences between Developmental Stages of Toxoplasma gondii Revealed by iTRAQ-Based Quantitative Proteomics.
Toxoplasma gondii has a complex two-host life-cycle between intermediate host and definitive host. Understanding proteomic variations across the life-cycle stages of T. gondii may improve the understanding of molecular adaption mechanism of T. gondii across life-cycle stages, and should have implications for the development of new treatment and prevention interventions against T. gondii infection. Here, we utilized LC-MS/MS coupled with iTRAQ labeling technology to identify differentially expressed proteins (DEPs) specific to tachyzoite (T), bradyzoites-containing cyst (C) and sporulated oocyst (O) stages of the cyst-forming T. gondii Prugniuad (Pru) strain. A total of 6285 proteins were identified in the three developmental stages of T. gondii. Our analysis also revealed 875, 656, and 538 DEPs in O vs. T, T vs. C, and C vs. O, respectively. The up- and down-regulated proteins were analyzed by Gene Ontology enrichment, KEGG pathway and STRING analyses. Some virulence-related factors and ribosomal proteins exhibited distinct expression patterns across the life-cycle stages. The virulence factors expressed in sporulated oocysts and the number of up-regulated virulence factors in the cyst stage were about twice as many as in tachyzoites. Of the 79 ribosomal proteins identified in T. gondii, the number of up-regulated ribosomal proteins was 33 and 46 in sporulated oocysts and cysts, respectively, compared with tachyzoites. These results support the hypothesis that oocyst and cystic stages are able to adapt to adverse environmental conditions and selection pressures induced by the host's immune response, respectively. These findings have important implications for understanding of the developmental biology of T. gondii, which may facilitate the discovery of novel therapeutic targets to better control toxoplasmosis.


Identification of the LIM protein FHL2 as a coactivator of beta-catenin.
Beta-catenin is a key mediator of the Wnt pathway, which plays a critical role in embryogenesis and oncogenesis. As a transcriptional activator, beta-catenin binds the transcription factors, T-cell factor and lymphoid enhancer factor, and regulates gene expression in response to Wnt signaling. Abnormal activation of beta-catenin has been linked to various types of cancer. In a yeast two-hybrid screen, we identified the four and a half of LIM-only protein 2 (FHL2) as a novel beta-catenin-interacting protein. Here we show specific interaction of FHL2 with beta-catenin, which requires the intact structure of FHL2 and armadillo repeats 1-9 of beta-catenin. FHL2 cooperated with beta-catenin to activate T-cell factor/lymphoid enhancer factor-dependent transcription from a synthetic reporter and the cyclin D1 and interleukin-8 promoters in kidney and colon cell lines. In contrast, coexpression of beta-catenin and FHL2 had no synergistic effect on androgen receptor-mediated transcription, whereas each of these two coactivators independently stimulated AR transcriptional activity. Thus, the ability of FHL2 to stimulate the trans-activating function of beta-catenin might be dependent on the promoter context. The detection of increased FHL2 expression in hepatoblastoma, a liver tumor harboring frequent beta-catenin mutations, suggests that FHL2 might enforce beta-catenin transactivation activity in cancer cells. These findings reveal a new function of the LIM coactivator FHL2 in transcriptional activation of Wnt-responsive genes.


Pulmonary infiltrates in immunocompromised patients: diagnosis by cytological examination of bronchoalveolar lavage fluid.
Thirty pulmonary infiltrates in 26 patients were investigated by bronchoalveolar lavage. Sixteen of the patients were on therapeutic immunosuppression for renal disease or transplant and 10 had leukaemia, lymphoma, or allied conditions. A rapid specific diagnosis was made in 21 (70%) episodes by cytological examination of the fluid and in 28 (93%) by a combination of cytology and microbiology. No complications from haemorrhage or pneumothorax ensued. Pneumonia due to Pneumocystis carinii was the most common diagnosis (27%), but opportunistic infections from cytomegalovirus, candida, aspergillus, zygomycetes, and acid fast bacilli were also identified by cytology. Two episodes were caused by occult pulmonary haemorrhage and five patients had malignant infiltration of the lung from leukaemia, myeloma, Hodgkin's disease, and lymphoplasmacytoid lymphoma. In two of these there was also evidence of infection. In seven cases with non-diagnostic cytology infections due to Staphylococcus aureus, Pseudomonas aeruginosa, pneumococcus, micrococcus, and Aspergillus fumigatus were identified on culture. In two patients (7%) no specific diagnosis was established by lavage: one had serological evidence of legionella infection and the second had P aeruginosa septicaemia. Twelve (75%) of the renal patients and six (60%) of those with leukaemia, lymphoma, and allied conditions recovered.


Adolescents and young adults' perceptions of electronic cigarettes as a gateway to smoking: a qualitative study in Switzerland.
Electronic cigarettes (ECs) acting as a gateway to smoking traditional cigarettes (TCs) is a growing public health concern of EC use among youths. To gather the opinions and perceptions of adolescents and young adults (AYAs) on whether and how EC can act as a gateway to smoking TC among youths. A qualitative method included 42 AYAs. Participants identified a significant risk of EC acting as a gateway to TC use, several factors contributing to this phenomenon such as a facilitated transition to smoking or its perception as a harmless toy. Participants considered an even greater gateway threat regarding very young adolescents. A minority did not identify the gateway risk and some believed that it was nicotine-dependent. This potential gateway effect brought forth several recommendations: health professionals should screen adolescents (even very young ones) for EC use and inform consumers of the potential gateway effect; this possible effect should be acknowledged to end the harmless perception many might have; there is an urge for better preventive and regulatory policies directed at protecting adolescents and children who never smoked and support those who have quit.