Joint Design with Providers of Clinical Decision Support for Value-Based Advanced Shoulder Imaging.
Provider orders for inappropriate advanced imaging, while rarely altering patient management, contribute enough to the strain on available health care resources, and therefore the United States Congress established the Appropriate Use Criteria Program. To examine whether co-designing clinical decision support (CDS) with referring providers will reduce barriers to adoption and facilitate more appropriate shoulder ultrasound (US) over magnetic resonance imaging (MRI) in diagnosing Veteran shoulder pain, given similar efficacies and only 5% MRI follow-up rate after shoulder US. We used a theory-driven, convergent parallel mixed-methods approach to prospectively (1) determine medical providers' reasons for selecting MRI over US in diagnosing shoulder pain and identify barriers to ordering US, (2) co-design CDS, informed by provider interviews, to prompt appropriate US use, and (3) assess CDS impact on shoulder imaging use. CDS effectiveness in guiding appropriate shoulder imaging was evaluated through monthly monitoring of ordering data at our quaternary care Veterans Hospital. Key outcome measures were appropriate MRI/US use rates and transition to ordering US by both musculoskeletal specialist and generalist providers. We assessed differences in ordering using a generalized estimating equations logistic regression model. We compared continuous measures using mixed effects analysis of variance with log-transformed data. During December 2016 to March 2018, 569 (395 MRI, 174 US) shoulder advanced imaging examinations were ordered by 111 providers. CDS "co-designed" in collaboration with providers increased US from 17% (58/335) to 50% (116/234) of all orders (p < 0.001), with concomitant decrease in MRI. Ordering appropriateness more than doubled from 31% (105/335) to 67% (157/234) following CDS (p < 0.001). Interviews confirmed that generalist providers want help in appropriately ordering advanced imaging. Partnering with medical providers to co-design CDS reduced barriers and prompted appropriate transition to US from MRI for shoulder pain diagnosis, promoting evidence-based practice. This approach can inform the development and implementation of other forms of CDS.


Visual mismatch negativity in the dorsal stream is independent of concurrent visual task difficulty.
The manipulation of attention can produce mismatch negativity-like components that are not necessarily connected to the unintentional sensory registration of the violation of probability-based regularity. For clinical purposes, attentional bias should be quantified because it can vary substantially among subjects and can decrease the specificity of the examination. This experiment targets the role of attention in the generation of visual mismatch negativity (vMMN). The visual regularity was generated by a sequence of two radial motions while subjects focused on visual tasks in the central part of the display. Attentional load was systematically varied and had three levels, no-load, easy, and difficult. Rare, deviant, and frequent standard motions were presented with a 10/60 ratio in oddball sequences. Data from 12 subjects was recorded from 64 channels and processed. vMMN was identified within the interval of 142-198 ms. The mean amplitude was evaluated during the aforementioned interval in the parietal and fronto-central regions. A general linear model for repeated measures was applied to the mean amplitude with a three-factor design and showed a significant difference [F (1, 11) = 17.40, p = 0.002] between standard and deviant stimuli and between regions [F (1, 11) = 8.40, p = 0.01]; however, no significant effect of the task [F (2, 22) = 1.26, p = 0.30] was observed. The unintentional detection of irregularity during the processing of the visual motion was independent of the attentional load associated with handling the central visual task. The experiment did not demonstrate an effect of attentional load manipulation on mismatch negativity (MMN) induced by the motion-sequence, which supports the clinical utility of this examination. However, used stimulation paradigm should be further optimized to generate mismatch negativity that is stable enough to be usable not only for group comparisons but also for a single subject assessment.


Seborrheic dermatitis: an overview.
Seborrheic dermatitis affects the scalp, central face, and anterior chest. In adolescents and adults, it often presents as scalp scaling (dandruff). Seborrheic dermatitis also may cause mild to marked erythema of the nasolabial fold, often with scaling. Stress can cause flare-ups. The scales are greasy, not dry, as commonly thought. An uncommon generalized form in infants may be linked to immunodeficiencies. Topical therapy primarily consists of antifungal agents and low-potency steroids. New topical calcineurin inhibitors (immunomodulators) sometimes are administered.


High Content Image Analysis as a Tool to Morphologically Distinguish Macrophage Activation and Determine Its Importance for Foamy Alveolar Macrophage Responses.
Lung diseases are an increasing global health burden affecting millions of people worldwide. Only a few new inhaled medicines have reached the market in the last 30 years, in part due to foamy alveolar macrophage (FAM) responses observed in pre-clinical rat studies. The induction mechanism and signaling pathways involved in the development of highly vacuolated 'foamy' phenotype is not known. Furthermore, it has not been determined if these observations are adaptive or adverse responses. To determine if high content image analysis techniques can distinguish between alveolar macrophage activation (LPS/IFN-γ activated and IL-4 activated macrophages) and if this could be applied to understanding the generation of 'foamy' macrophage phenotypes. NR8383 rat alveolar macrophages were stimulated with a mix of cytokines (LPS/IFN-γ or IL-4) for 24 h. The cells were further exposed to FAM inducing-compounds amiodarone and staurosporine. Following 24 h incubation, phagocytosis and lipid accumulation were measured using flow cytometry and high content image analysis techniques. The alveolar macrophages responses after exposure to cytokines were assessed by evaluation: (i) cell surface and biochemical markers such as: nitric oxide production, arginase-1 activity and MRC-1 receptor expression (ii) cellular morphology (iii) cellular functionality (phagocytic activity and lipids accumulation). Macrophages activated with LPS/IFN-γ showed distinct morphological (increased vacuolation) features and functionality (increased lipidosis, decreased phagocytic activity). Foamy macrophage phenotypes induced by amiodarone also displayed characteristics of proinflammatory macrophages (significantly increased nitric oxide production, increased vacuolation and lipidosis and decreased phagocytosis). In contrast, staurosporine treatment resulted in increased NO production, as well as arginase-1 activity. High content image analysis was able to determine distinct differences in morphology between non-activated and LPS/IFN-γ activated macrophages, characterized by increased vacuolation and lipidosis. When exposed to compounds that induce a FAM phenotype, healthy non-activated macrophages displayed proinflammatory (amiodarone) or pro-apoptotic (staurosporine) characteristics but these responses were independent of a change in activation status. This technique could be applied in early drug discovery safety assessment to identify immune responses earlier and increase the understanding of alveolar macrophage responses to new molecules challenge in development of new inhalation therapies, which in turn will enhance decision-making in an early safety assessment of novel drug candidates.


Increasing rural-urban gradients in US suicide mortality, 1970-1997.
This study examined rural-urban gradients in US suicide mortality and the extent to which such gradients varied across time, sex, and age. Using a 10-category rural-urban continuum measure and 1970-1997 county mortality data, we estimated rural-urban differentials in suicide mortality over time by multiple regression and Poisson regression models. Significant rural-urban gradients in age-adjusted male suicide mortality were found in each time period, indicating rising suicide rates with increasing levels of rurality. The gradient increased consistently, suggesting widening rural-urban differentials in male suicides over time. When controlled for geographic variation in divorce rate and ethnic composition, rural men, in each age cohort, had about twice the suicide rate of their most urban counterparts. Observed rural-urban differentials for women diminished over time. In 1995 to 1997, the adjusted suicide rates for young and working-age women were 85% and 22% higher, respectively, in rural than in the most urban areas. The slope of the relationship between rural-urban continuum and suicide mortality varied substantially by time, sex, and age. Widening rural-urban disparities in suicide may reflect differential changes over time in key social integration indicators.


Calcimimetic Use in Familial Hypocalciuric Hypercalcemia-A Perspective in Endocrinology.
Familial hypocalciuric hypercalcemia (FHH) causes lifelong hypercalcemia that even persists after subtotal parathyroidectomy. Symptoms are usually mild. Past recommendations have often been for monitoring and against surgical or pharmacologic treatments. Review of publications about FHH, calcium-sensing receptors (CaSRs), and calcimimetics. FHH reflects heterozygous germline mutation of CASR, GNA11, or AP2S1. These mutations inactivate the CaSRs in the parathyroid cell. Thereby, they shift the serum calcium set point to higher values and cause hypercalcemia. Calcimimetic drugs enhance the effects of calcium on the CaSRs and thereby inhibit the parathyroid cell. Calcimimetic drugs are indicated in adults with primary hyperparathyroidism without a good surgical option. Calcimimetic safety and efficacy are not established in children younger than age 18 years. Recent case reports have described treatment of FHH with calcimimetics. Success was classified as combinations of subjective improvements and decreases of serum calcium levels, but not necessarily into the normal range. Treatment was successful in 14 of 16 cases (88%). Deductions based on these case reports have limitations. For example, failures of therapy may not have been reported. Cost of the drug might be rate limiting. Calcimimetics can be offered to adults with FHH and those in whom the serum calcium level is >0.25 mM (1 mg/dL) beyond the upper limit of normal or with possible symptoms of hypercalcemia. Calcimimetics can now be offered to more adults with FHH.


Infectious bursal disease virus influences the transcription of chicken γc and γc family cytokines during infection.
Infectious bursal disease virus (IBDV) infection causes immunodeficiency in chickens. To understand cell-mediated immunity during IBDV infection, this study perform a detailed analysis of chicken γc chain (chCD132) and γc family cytokines, including interleukins 2, 4, 7, 9, and 15. The mouse anti-chCD132 monoclonal antibody (mAb) was first generated by the E.coli-expressed γc protein. Immunofluorescence assay further showed that γc was a protein located with the anti-chCD132 mAb on the surface of chicken's splenic mononuclear cells. Real-time quantitative RT-PCR revealed that the chCD132 mRNA transcript was persistently downregulated in embryo fibroblasts, spleen and thymus of chickens infected with IBDV. Correspondingly during IBDV infection, the transcription of five γc family cytokines was downregulated in the thymus and presented an imbalance in the spleen. Fluorescence-activated cell sorting analyses also indicated that the percentage of CD132(+)CD8(+) T cells linearly decreased in the bursa of IBDV-infected chickens. These results confirmed that IBDV infection disturbed the in vivo balance of CD132 and γc family cytokine expression and that IBDV-induced immunodeficiency involved cellular networks related to the γc family.


[Pregnancy after isotretinoin use].
Acne is a common problem that is increasingly being effectively treated with isotretinoin, a highly teratogenic agent. According to the pregnancy prevention program, contraception should be used until at least one month after discontinuing the medication. A 29-year-old woman became pregnant shortly after discontinuing isotretinoin. She was referred to the gynaecologist for an abortion. However, since she wished to continue the pregnancy, she received personal advice on the risk of bearing a child with congenital abnormalities. After an uneventful pregnancy she gave birth to a healthy son. Given the teratogenicity of isotretinoin, every woman in the fertile phase of life should use contraception while taking the medication. If she becomes pregnant, however, the individual risk of having a child with congenital abnormalities can be carefully and reliably estimated by determining the precise date of conception through ultrasound and the date that isotretinoin was stopped.


Impaired sperm maturation in conditional Lcn6 knockout mice.
Human LCN6, a lipocalin protein, exhibits predominant expression in epididymis and location on the sperm surface. However, the biological function of LCN6 in vivo remains unknown. Herein, we found that unlike human LCN6, mouse Lcn6 gene encoded two transcript variants that were both upregulated by androgen. Subsequently, we generated a conditional knockout mouse model to disrupt Lcn6 in the adult and investigate its function. In this model, spermatogenesis was normal and Lcn6 deficiency did not affect the natural birth rate of male mice or in vitro fertilization ability of their cauda epididymal sperm. Nevertheless, sperm from the cauda epididymis of the Lcn6 null mice underwent a sustained increase of acrosome reaction frequency whether capacitated or not (P < 0.01). Consistent with premature acrosome reaction, sperm from knockout mice had significantly increased intracellular calcium content when extracellular calcium was supplied (P < 0.01). These results demonstrate an important function of LCN6 in preventing calcium overload and premature acrosome reaction of sperm and suggest a potential risk factor of LCN6 deficiency for sperm maturation.


A genome-wide investigation of copy number variation in patients with sporadic brain arteriovenous malformation.
Brain arteriovenous malformations (BAVM) are clusters of abnormal blood vessels, with shunting of blood from the arterial to venous circulation and a high risk of rupture and intracranial hemorrhage. Most BAVMs are sporadic, but also occur in patients with Hereditary Hemorrhagic Telangiectasia, a Mendelian disorder caused by mutations in genes in the transforming growth factor beta (TGFβ) signaling pathway. To investigate whether copy number variations (CNVs) contribute to risk of sporadic BAVM, we performed a genome-wide association study in 371 sporadic BAVM cases and 563 healthy controls, all Caucasian. Cases and controls were genotyped using the Affymetrix 6.0 array. CNVs were called using the PennCNV and Birdsuite algorithms and analyzed via segment-based and gene-based approaches. Common and rare CNVs were evaluated for association with BAVM. A CNV region on 1p36.13, containing the neuroblastoma breakpoint family, member 1 gene (NBPF1), was significantly enriched with duplications in BAVM cases compared to controls (P = 2.2×10(-9)); NBPF1 was also significantly associated with BAVM in gene-based analysis using both PennCNV and Birdsuite. We experimentally validated the 1p36.13 duplication; however, the association did not replicate in an independent cohort of 184 sporadic BAVM cases and 182 controls (OR = 0.81, P = 0.8). Rare CNV analysis did not identify genes significantly associated with BAVM. We did not identify common CNVs associated with sporadic BAVM that replicated in an independent cohort. Replication in larger cohorts is required to elucidate the possible role of common or rare CNVs in BAVM pathogenesis.


Spontaneous rupture of the spleen with EMATOMA.
A case of spontaneous rupture of the spleen with hematoma is presented. The spleen presumably with ruptured while the patient was in the hospital, during a severe coughing episode secondary to viral pneumonia. The time interval between splenic rupture and operation was 31 days. Along with pertinent diagnostic acids, difficulty in making the diagnosis is stressed. Knowledge that this condition exists is a major factor in making the diagnosis.


Tobacco-Nicotine Education and Training for Health-Care Professional Students and Practitioners: A Systematic Review.
The objective of this systematic review was to investigate what education and training characteristics prepares and supports health-care professionals (HCPs) in the delivery of competent and effective care to clients who use tobacco-nicotine. A search of eight bibliographic databases for English-language peer-reviewed publications from January 2006 to March 2015. Studies were included if they met the a priori inclusion criteria, which consisted of: (1) quantitative study design and (2) focus on tobacco-nicotine education or training for HCP students and practitioners. All studies were independently screened for inclusion by two reviewers. Data from included studies were extracted for study characteristics and key outcomes then critically appraised for methodological quality. Fifty-nine studies were included for narrative synthesis. Two categories emerged: (1) curriculum characteristics (n = 10) and (2) education and training interventions (n = 49). Included curriculum studies identified the following themes: content, intensity, competencies evaluation, and barriers. Study findings about education and training interventions were grouped by level of education (prelicensure, post-licensure, and faculty training), teaching modality, health discipline, and the associated HCP and client outcomes. This comprehensive review suggests that there is a lack of consistency in HCP tobacco-nicotine education and training characteristics. This paper provides valuable categorization of the most frequently utilized components of academic curriculum and discusses the interventions in relation to HCP and client outcomes. Gaps in the literature are highlighted, and the need for standardization of tobacco-nicotine training competencies and evaluation is discussed. Future research investigating the most effective approaches to training is needed. This systematic review summarizes existing tobacco-related curriculum components (content, intensity, competency evaluation, and barriers) and training interventions for health-care professionals worldwide and demonstrates that they are associated with positive health-care professional outcomes (knowledge, attitudes, behaviors, and skills) and client outcomes (quit attempts and smoking abstinence).


Estimation of distance related probability of animal movements between holdings and implications for disease spread modeling.
Between holding contacts are more common over short distances and this may have implications for the dynamics of disease spread through these contacts. A reliable estimation of how contacts depend on distance is therefore important when modeling livestock diseases. In this study, we have developed a method for analyzing distant dependent contacts and applied it to animal movement data from Sweden. The data were analyzed with two competing models. The first model assumes that contacts arise from a purely distance dependent process. The second is a mixture model and assumes that, in addition, some contacts arise independent of distance. Parameters were estimated with a Bayesian Markov Chain Monte Carlo (MCMC) approach and the model probabilities were compared. We also investigated possible between model differences in predicted contact structures, using a collection of network measures. We found that the mixture model was a much better model for the data analyzed. Also, the network measures showed that the models differed considerably in predictions of contact structures, which is expected to be important for disease spread dynamics. We conclude that a model with contacts being both dependent on, and independent of, distance was preferred for modeling the example animal movement contact data.


Single-channel recordings of three K+-selective currents in cultured chick ciliary ganglion neurons.
Multiple distinct K+-selective channels may contribute to action potential repolarization and afterpotential generation in chick ciliary neurons. The channel types are difficult to distinguish by traditional voltage-clamp methods, primarily because of coactivation during depolarization. I have used the extracellular patch-clamp technique to resolve single-channel K+ currents in cultured chick ciliary ganglion (CG) neurons. Three unit currents selective for K+ ions were observed. The channels varied with respect to unit conductance, sensitivity to Ca2+ ions and voltage, and steady-state gating parameters. The first channel, GK1, was characterized by a unit conductance of 14 pico-Siemens (pS) under physiological recording conditions, gating that was relatively independent of membrane potential and intracellular Ca2+ ions, and single-component open-time distributions with time constants of approximately 9 msec. The second channel, GK2, was characterized by a unit conductance of 64 pS under physiological recording conditions and gating that was affected by membrane potential but was not dependent on the activity of intracellular Ca2+ ions. Open-time distributions indicated 2 open states, with open-time constants of 0.09 (61%) and 0.35 (39%) msec, at +40 mV membrane potential. The third channel, GKCa2+, was identified in isolated patch recordings in which the concentration of internal Ca2+ was 10(-7) M or greater, which was an absolute prerequisite for channel opening. GKCa2+ was characterized by a unit conductance of 193 pS in symmetrical 0.15 M KCl solutions, an open-state probability that was a function not only of [Ca2+]i, but also of membrane potential, and single-component open-time distribution with a time constant of 1.11 msec at -10 mV patch potential. These results suggest the presence of at least 3 distinct K+ channel populations in the membrane of cultured chick CG neurons.


Construction and biological characterization of a cDNA infectious clone of wheat umbra-like virus in wheat and Nicotiana benthamiana.
Umbravirus-like associated RNAs (ulaRNAs) are a new group of subviral RNAs associated with plants. Little is known about the biology of ulaRNAs. We recently reported wheat umbra-like virus (WULV) from Kansas fields. In this work, we generated a full-length cDNA clone of WULV which systemically infected N. benthamiana. While agroinfiltrated leaves demonstrated severe necrosis, upper leaves were symptomless. We also showed that WULV is capable of infecting wheat in the absence of a helper virus. Furthermore, and through sap inoculation, we demonstrated that WULV is transmissible in the form of free RNA. This is the first report of a mechanically transmissible ulaRNA. Together, these findings contribute to advancing our knowledge of the biology of WULV. Moreover, the construction of the WULV infectious clone provides a valuable research tool for further investigations including the role of WULV in symptom development in interaction with other wheat viruses.


Clinical histopathological characteristics of basal cell carcinoma in Japanese patients.
Basal cell carcinoma (BCC) does not commonly occur in the African and Asian races. Thus, there have been few reviews that have described the clinical overview of BCC among Japanese patients. There have been no large-scale analyses, including the histopathological features of BCC, except for the white race. We retrospectively examined 243 Japanese patients with BCC and analyzed the color of the tumors, the patient's age at which these tumors occurred, the site of the lesions, the histopathological patterns, and the incidence of recurrence and metastasis of the tumors; we compared these features with those in the other races statistically. Clinically, approximately 75% of the tumors were pigmented and showed a so-called black pearly appearance. The male-female ratio was 0.97, and the average age of the patients was 59 years. Seventy-five percent of the tumors occurred on the head and neck. Fifty-four percent of the tumors showed a solid type of histopathological pattern. The incidence of recurrence and metastasis is extremely rare. The high incidence of hyperpigmentation in the lesional skin of BCC is the most characteristic feature of BCCs in Japanese patients.


Human energy - optimal control of disturbance rejection during constrained standing.
An optimal control system that enables a subject to stand without hand support in the sagittal plane was designed. The subject was considered as a double inverted pendulum structure with a voluntarily controlled degree of freedom in the upper trunk and artificially controlled degree of freedom in the ankle joints. The control system design was based on a minimization of cost function that estimated the effort of the ankle joint muscles through observation of the ground reaction force position relative to the ankle joint axis. By maintaining the centre of pressure close to the ankle joint axis the objective of the upright stance is fulfilled with minimal ankle muscle energy cost. The performance of the developed controller was evaluated in a simulation-based study. The results were compared with the responses of an unimpaired subject to different disturbances in the sagittal plane. The proposed cost function was shown to produce a reasonable approximation of human natural behaviour.


Induction of p53 by urokinase in lung epithelial cells.
Urokinase plasminogen activator (uPA) is a serine protease that catalyzes the conversion of plasminogen to plasmin. The plasminogen/plasmin system includes the uPA, its receptor, and its inhibitor (plasminogen activator inhibitor-1). Interactions between these molecules regulate cellular proteolysis as well as adhesion, cellular proliferation, and migration, processes germane to the pathogenesis of lung injury and neoplasia. In previous studies, we found that uPA regulates cell surface fibrinolysis by regulating its own expression as well as that of the uPA receptor and plasminogen activator inhibitor-1. In this study, we found that uPA alters expression of the tumor suppressor protein p53 in Beas2B airway epithelial cells in both a time- and concentration-dependent manner. These effects do not require uPA catalytic activity because the amino-terminal fragment of uPA lacking catalytic activity was as potent as two chain active uPA. Single chain uPA also enhanced p53 expression to the same extent as intact two chain active uPA and the amino-terminal fragment. Pretreatment of cells with anti-beta1 integrin antibody blocked uPA-induced p53 expression. uPA-induced p53 expression occurs without increased p53 mRNA expression. However, uPA induced oncoprotein MDM2 in a concentration-dependent manner. uPA-induced p53 expression does not require activation of tyrosine kinases. Inactivation of protein-tyrosine phosphatase SHP-2 inhibits both basal and uPA-induced p53 expression. Plasmin did not alter uPA-mediated p53 expression. The induction of p53 expression by exposure of lung epithelial cells to uPA is a newly recognized pathway by which urokinase may influence the proliferation of lung epithelial cells. This pathway could regulate pathophysiologic alterations of p53 expression in the setting of lung inflammation or neoplasia.


Aryl hydrocarbon receptor modulates NADPH oxidase activity via direct transcriptional regulation of p40phox expression.
A member of the NADPH oxidase subunits, p40(phox) plays an important role in the regulation of NADPH oxidase activity and the subsequent production of reactive oxygen species (ROS). In this study, we show that mouse p40(phox) is a novel transcriptional target of the aryl hydrocarbon receptor (AhR), known as a dioxin receptor or xenobiotic receptor, in the liver. Treatment of mice with 3-methylcholanthrene (3MC) increased p40(phox) gene expression in the liver, but this induction of p40(phox) gene expression was diminished by the deletion of the AhR gene in the liver. Consistent with the in vivo results, the expression of the p40(phox) gene was increased in 3MC-treated Hepa1c1c7 cells in an AhR-dependent manner. In addition, promoter analysis established p40(phox) as a transcriptional target of AhR. Studies using the RNA-interference technique revealed that p40(phox) is involved in the increase of NADPH oxidase activity and the subsequent ROS production in AhR-activated Hepa1c1c7 cells. Consequently, the results obtained here may provide a novel molecular mechanism for ROS production after exposure to dioxins.


HIV-associated nephropathy: outcome and prognosis factors. Groupe d' Etudes Néphrologiques d'Ile de France.
Records of 102 patients with biopsy-proven HIV-associated nephropathy (HIVAN) admitted to 18 hospitals in the Paris area from 1984 through 1996 were retrospectively reviewed. Demographics and clinical and laboratory features of the cohort were determined, and prognostic factors of renal and patient survival were analyzed. Renal and patient survival curves were estimated with the actuarial method. Prognostic factors were assessed by uni- and multidimensional analyses based on Cox regression models. Values were expressed as median with interquartile. The total population (median age 34) included 97% blacks and 71.5% males. Median patient follow-up was 165 d (range, 43 to 493). At the time of renal biopsy, median values of serum creatinine, proteinuria, and CD4+ cell count were 496 micromol/L, 6.5 g/24 h, and 48.5 cells/mm3, respectively. Fifteen patients were given steroids after the onset of HIVAN. Overall patient survival at 0.5, 1, and 3 yr was 73 +/- 5, 55 +/- 6, and 38 +/- 7%, respectively. The proportion of patients free of dialysis at 0.5, 1, and 3 yr was 73 +/- 5, 60 +/- 7, and 18 +/- 10%, respectively. Predictors of poor patient prognosis were a low CD4+ cell count (relative risk [RR; per 50 cells/mm3 decrease] 1.35; confidence interval [CI], 1.13 to 1.6) and antiretroviral therapy before the onset of HIVAN (RR 1.9; CI, 1.05 to 3.6). Main independent factors associated with better renal outcome were: steroid therapy (RR 0.29; CI, 0.1 to 0.9); low proteinuria level (RR [per 50% decrease] 0.7; CI, 0.5 to 0.98); low serum creatinine (RR [per 1.1 mg/dl decrease] 0.78; CI, 0.7 to 0.87); and hemoglobin level (RR [per g/dl increase] 0.76; CI, 0.58 to 1.00). HIVAN is not a rare nephropathy in Paris and its suburbs. Renal prognosis and patient survival are better than what was reported previously. Steroids may delay the downward course of HIVAN. It is not certain that in the new era of HIV therapy, the possible renal benefits of corticosteroids outweigh their potential risks. The only reliable predictor of patient survival is the intensity of immunodeficiency.


Agrobacterium tumefaciens mediated transformation of the aquatic carnivorous plant Utricularia gibba.
The genus Utricularia belongs to Lentibulariaceae, the largest family of carnivorous plants, which includes terrestrial, epiphytic and aquatic species. The development of specialized structures that evolved for carnivory is a feature of this genus that has been of great interest to biologists since Darwin's early studies. Utricularia gibba is itself an aquatic plant with sophisticated bladder traps having one of the most complex suction mechanisms for trapping prey. However, the molecular characterization of the mechanisms that regulate trap development and the biophysical processes involved in prey trapping are still largely unknown due to the lack of a simple and reproducible gene transfer system. Here, we report the establishment of a simple, fast and reproducible protocol for genetic transformation of U. gibba based on the T-DNA of Agrobacterium tumefaciens. An in vitro selection system using Phosphinotricin as a selective agent was established for U. gibba. Plant transformation was confirmed by histochemical GUS assays and PCR and qRT-PCR analyses. We report on the expression pattern of the 35S promoter and of the promoter of a trap-specific ribonuclease gene in transgenic U. gibba plants. The genetic transformation protocol reported here is an effective method for studying developmental biology and functional genomics of this genus of carnivorous plants and advances the utility of U. gibba as a model system to study developmental processes involved in trap formation.


Acoustic droplet vaporization is initiated by superharmonic focusing.
Acoustically sensitive emulsion droplets composed of a liquid perfluorocarbon have the potential to be a highly efficient system for local drug delivery, embolotherapy, or for tumor imaging. The physical mechanisms underlying the acoustic activation of these phase-change emulsions into a bubbly dispersion, termed acoustic droplet vaporization, have not been well understood. The droplets have a very high activation threshold; its frequency dependence does not comply with homogeneous nucleation theory and localized nucleation spots have been observed. Here we show that acoustic droplet vaporization is initiated by a combination of two phenomena: highly nonlinear distortion of the acoustic wave before it hits the droplet and focusing of the distorted wave by the droplet itself. At high excitation pressures, nonlinear distortion causes significant superharmonics with wavelengths of the order of the droplet size. These superharmonics strongly contribute to the focusing effect; therefore, the proposed mechanism also explains the observed pressure thresholding effect. Our interpretation is validated with experimental data captured with an ultrahigh-speed camera on the positions of the nucleation spots, where we find excellent agreement with the theoretical prediction. Moreover, the presented mechanism explains the hitherto counterintuitive dependence of the nucleation threshold on the ultrasound frequency. The physical insight allows for the optimization of acoustic droplet vaporization for therapeutic applications, in particular with respect to the acoustic pressures required for activation, thereby minimizing the negative bioeffects associated with the use of high-intensity ultrasound.


[Health care access and receptivity to users in a unit in Porto Alegre, Rio Grande do Sul, Brazil].
This study focuses on users' views of factors influencing quality of care at a health care unit in the city of Porto Alegre, relating to access and receptivity. The data were collected using a semi-structured interview and participatory observation and treated using thematic analysis. The results compare ease and difficulties in geographic, economic, and functional access. Organization of services and professional competency were determinant factors in ease of reception, leading to user satisfaction. Poor reception and unsatisfactory professional performance were identified as difficulties. The study concluded that there is a need to increase the professional staff, train them in receiving users, implement a complementary modality for dental care, open the facility earlier for scheduling appointments, and prioritize care for residents of the catchment area.


European consensus-based recommendations for the diagnosis and treatment of rare paediatric vasculitides - the SHARE initiative.
The European initiative Single Hub and Access point for paediatric Rheumatology in Europe (SHARE) aimed to optimize care for children with rheumatic diseases. Systemic vasculitides are very rare in children. Consequently, despite recent advances, paediatric-specific information is sparse. The lack of evidence-based recommendations is an important, unmet need. This study aimed to provide recommendations for diagnosing and treating children with rare forms of childhood systemic vasculitis. Recommendations were developed by a consensus process in accordance with the European League Against Rheumatism standard operating procedures. A systematic literature review informed the recommendations, which were devised and evaluated by a panel of experts via an online survey, and two consensus meetings using nominal group technique. Recommendations were accepted when ⩾ 80% of experts agreed. Ninety-three relevant articles were found, and 78 recommendations were accepted in the two consensus meetings. General, cross-cutting recommendations and disease-specific statements regarding the diagnosis and treatment of childhood-onset PAN, granulomatosis with polyangiitis, microscopic polyangiitis, eosinophilic granulomatosis with polyangiitis, and Takayasu arteritis are provided. These Single Hub and Access point for paediatric Rheumatology in Europe recommendations were formulated through an evidence-based consensus process to support uniform, high-quality standard of care for children with rare forms of paediatric systemic vasculitis.


Gene expression profiling of Burkholderia cenocepacia at the time of cepacia syndrome: loss of motility as a marker of poor prognosis?
Cepacia syndrome (CS) is a fatal septic condition that develops in approximately 20% of cystic fibrosis (CF) patients chronically infected with the Burkholderia cepacia complex (Bcc). The most common causative agent is Burkholderia cenocepacia, a clinically dominant Bcc species that contains the globally distributed epidemic strain sequence type 32 (ST32). Using microarrays, we compared the transcriptomes of ST32 isolates from the bloodstream at the time of CS with their sputum counterparts recovered 1 to 2 months prior to the development of CS. Global gene expression profiles of blood isolates revealed greater activities of the virulence genes involved in the type III secretion system, the bacterial exopolysaccharide cepacian, and quorum sensing, while reduced expression was demonstrated for flagellar genes. Furthermore, a nonmotile phenotype (as evaluated by a swimming motility assay) was identified in blood isolates from 6 out of 8 patients with CS; this phenotype was traceable to 24 months prior to the onset of CS. Loss of motility was not observed in any of the 89 ST32 isolates recovered over the course of chronic infection from 17 patients without CS. In conclusion, the gene expression of Bcc bacteria disseminated during CS has been elucidated for the first time. This study demonstrated marked differences at the transcriptome level between isogenic ST32 isolates that are attributable to the stage and site of infection. The finding of a nonmotile B. cenocepacia isolate may serve as a warning sign for the development of CS in the near future.


Trauma Non-Technical Training (TNT-2): the development, piloting and multilevel assessment of a simulation-based, interprofessional curriculum for team-based trauma resuscitation.
Medical error is common during trauma resuscitations. Most errors are nontechnical, stemming from ineffective team leadership, nonstandardized communication among team members, lack of global situational awareness, poor use of resources and inappropriate triage and prioritization. We developed an interprofessional, simulation-based trauma team training curriculum for Canadian surgical trainees. Here we discuss its piloting and evaluation.


Talc-induced inflammation in the pleural cavity.
Talc administration into the pleural cavity induces pleurodesis. To obtain further insight into the inflammatory process that causes pleurodesis, the cellular kinetics in the pleural space after the administration of talc was studied, along with its relation to chemokine concentrations in the pleural fluid. Thirteen consecutive patients with idiopathic spontaneous pneumothorax and eight patients with malignant pleural effusions received talc pleurodesis. The first group was treated with talc poudrage, whereas the second group was treated with talc slurry. Pleural fluids were isolated before talc administration as well as 3, 6, 24, 48 and 72 h afterwards. The talc induced a rapid polymorphonuclear neutrophil (PMN) influx followed by an accumulation of macrophages. In addition, increased production of interleukin (IL)-8 and monocyte chemotactic protein (MCP)-1 was observed. The talc-induced PMN influx reached its maximum after 3-24 h, and was related to the IL-8 concentration. In contrast, the MCP-1 was not related to the macrophage accumulation. Talc-induced inflammation in patients with idiopathic spontaneous pneumothorax and malignant pleural effusion is characterized by an influx of polymorphonuclear neutrophils related to interleukin-8, followed by an accumulation of monocytes.


Experimental evidence that source genetic variation drives pathogen emergence.
A pathogen can readily mutate to infect new host types, but this does not guarantee successful establishment in the new habitat. What factors, then, dictate emergence success? One possibility is that the pathogen population cannot sustain itself on the new host type (i.e. host is a sink), but migration from a source population allows adaptive sustainability and eventual emergence by delivering beneficial mutations sampled from the source's standing genetic variation. This idea is relevant regardless of whether the sink host is truly novel (host shift) or whether the sink is an existing or related, similar host population thriving under conditions unfavourable to pathogen persistence (range expansion). We predicted that sink adaptation should occur faster under range expansion than during a host shift owing to the effects of source genetic variation on pathogen adaptability in the sink. Under range expansion, source migration should benefit emergence in the sink because selection acting on source and sink populations is likely to be congruent. By contrast, during host shifts, source migration is likely to disrupt emergence in the sink owing to uncorrelated selection or performance tradeoffs across host types. We tested this hypothesis by evolving bacteriophage populations on novel host bacteria under sink conditions, while manipulating emergence via host shift versus range expansion. Controls examined sink adaptation when unevolved founding genotypes served as migrants. As predicted, adaptability was fastest under range expansion, and controls did not adapt. Large, similar and similarly timed increases in fitness were observed in the host-shift populations, despite declines in mean fitness of immigrants through time. These results suggest that source populations are the origin of mutations that drive adaptive emergence at the edge of a pathogen's ecological or geographical range.


Suitability and Potential Nutrient Contribution of Underutilized Foods in Community-Based Infant Foods in Northern Ghana.
In rural Ghana, infant feeding is largely home-based or community-based yet less is known about the kinds of community-based infant foods and the ability of families to create a range of recipes for baby feeding using context-specific ingredients particularly in northern Ghana which has a high prevalence of malnutrition. In this explorative study on mothers (15-49 years; n = 46), we investigated community-based infant foods' food group composition, enrichment, nutrient contribution, and acceptability. The identified community-based infant foods were mainly made of either corn or millet porridges in northern Ghana and had three nutrients with % RNI ≥ 70. We developed 38 recipes of enriched community-based infant foods adding underutilized foods (orange-fleshed sweet potato, pawpaw, cowpea, moringa, groundnut, Bambara beans, and soya beans) to increase the number of nutrients from three to at least five and at most nine nutrients with % RNI ≥ 70 based on the recipe combinations. The enriched community-based infant food recipes provided adequate caloric amounts and modest improvements in micronutrient content for infants (6-12 months). All recipes tested were deemed appropriate and acceptable for infants by mothers. Moringa and pawpaw emerged as the lowest-cost ingredients to add among the underutilized foods. Future research is necessary to assess the effectiveness of the new recipes at promoting linear growth and improving micronutrient status during the complementary feeding period.


Long-term survival following radiofrequency ablation of colorectal liver metastases: A retrospective study.
To retrospectively evaluate the long-term survival of patients that received radiofrequency ablation (RFA) therapies of colorectal liver metastases. In 2005 to 2008, RFA of 105 colorectal liver metastases (CRLM) were performed on 49 patients in our institution. The liver metastases were evaluated, both before and after ablation therapies, with contrast enhanced computerised tomography and contrast enhanced ultrasonography. Histological evidence of malignant liver metastases was obtained in the few instances where contrast enhanced ultrasonography gave equivocal results. Accesses to the CRLM were guided ultrasonically in all patients. The data obtained from records of these ablations were retrospectively analysed and survival data were compared with existing studies in the literature. 1-, 2-, 3-, 4- and 5-year survival rates, when no stringent selection criteria were applied, were 92%, 65%, 51%, 41% and 29% respectively. To explore the impact of the number and size of CRLM on patients' survival, an exclusion of 13 patients (26.5%) with number of CRLM ≥ 5 and tumour size ≥ 40 mm resulted in 1-, 2-, 3-, 4- and 5-year survival rates improving to 94%, 69%, 53%, 42% and 31% respectively. It is of note that 9 of 49 patients developed extra-hepatic metastases, not visible or seen on pre-treatment scans, just after RFA treatment. These patients had poorer survival. The development of extra-hepatic metastases in nearly 20% of the patients included in our study can partly account for modestly lower survival rates as compared with earlier studies in the literature. Our study underscores the fact that optimum patients' selection before embarking on RFA treatment is vitally important to achieving a superior outcome.


Time-frequency spectral analysis of TMS-evoked EEG oscillations by means of Hilbert-Huang transform.
A single pulse of Transcranial Magnetic Stimulation (TMS) generates electroencephalogram (EEG) oscillations that are thought to reflect intrinsic properties of the stimulated cortical area and its fast interactions with other cortical areas. Thus, a tool to decompose TMS-evoked oscillations in the time-frequency domain on a millisecond timescale and on a broadband frequency range may help to understand information transfer across cortical oscillators. Some recent studies have employed algorithms based on the Wavelet Transform (WT) to study TMS-evoked EEG oscillations in healthy and pathological conditions. However, these methods do not allow to describe TMS-evoked EEG oscillations with high resolution in time and frequency domains simultaneously. Here, we first develop an algorithm based on Hilbert-Huang Transform (HHT) to compute statistically significant time-frequency spectra of TMS-evoked EEG oscillations on a single trial basis. Then, we compared the performances of the HHT-based algorithm with the WT-based one by applying both of them to a set of simulated signals. Finally, we applied both algorithms to real TMS-evoked potentials recorded in healthy or schizophrenic subjects. We found that the HHT-based algorithm outperforms the WT-based one in detecting the time onset of TMS-evoked oscillations in the classical EEG bands. These results suggest that the HHT-based algorithm may be used to study the communication between different cortical oscillators on a fine time scale.


Structural Elucidation and Biological Activity of a Highly Regular Fucosylated Glycosaminoglycan from the Edible Sea Cucumber Stichopus herrmanni.
Edible sea cucumbers are widely used as a health food and medicine. A fucosylated glycosaminoglycan (FG) was purified from the high-value sea cucumber Stichopus herrmanni. Its physicochemical properties and structure were analyzed and characterized by chemical and instrumental methods. Chemical analysis indicated that this FG with a molecular weight of ∼64 kDa is composed of N-acetyl-d-galactosamine, d-glucuronic acid (GlcA), and l-fucose. Structural analysis clarified that the FG contains the chondroitin sulfate E-like backbone, with mostly 2,4-di-O-sulfated (85%) and some 3,4-di-O-sulfated (10%) and 4-O-sulfated (5%) fucose side chains that link to the C3 position of GlcA. This FG is structurally highly regular and homogeneous, differing from the FGs of other sea cucumbers, for its sulfation patterns are simpler. Biological activity assays indicated that it is a strong anticoagulant, inhibiting thrombin and intrinsic factor Xase. Our results expand the knowledge on structural types of FG and illustrate its biological activity as a functional food material.


Enhanced osseointegration of titanium implant through the local delivery of transcription factor SATB2.
Titanium implants are widely used in dentistry and orthopedic surgery. Nevertheless, bone regeneration around the implant is a relatively slow process, after placement. This study assessed whether SATB2 can enhance osseointegration of a titanium implant. To determine the effect of SATB2 in implant integration, two different viruses encoding SATB2 (PBABE-Satb2 virus or RCAS-Satb2 virus) were locally administered to the bone defect prior to titanium implant placement in our established transgenic TVA mice. Seven and 21 days post implantation, the femurs were isolated for quantitative real-time RT-PCR, H&E staining, immunohistochemical (IHC) staining, and microcomputed tomography (microCT) analysis. Quantitative real-time RT-PCR results demonstrated that the in vivo overexpression of SATB2 enhanced expression levels of potent osteogenic transcription factors and bone matrix proteins. We also found that 21 days after implantation, there were no significant differences in the expression levels of SATB2, Osx, Runx2, COLI, OC, and BSP between the RCAS-Satb2 group and the RCAS group. Histological analysis showed that SATB2 overexpression significantly enhanced new bone formation and bone-to-implant contact after implantation. IHC staining analysis revealed that forced expression of SATB2 increased the number of BSP-positive cells surrounding the implant. MicroCT analysis demonstrated that in vivo overexpression of SATB2 significantly increased the density of the newly formed bone surrounding the implant. These results conclude that in vivo overexpression of SATB2 significantly accelerates osseointegration of titanium implants and SATB2 can serve as a potent molecule in promoting tissue regeneration.


Role of nerve growth factor in the development of rat sympathetic neurons in vitro. I. Survival, growth, and differentiation of catecholamine production.
To study the effect of nerve growth factor (NGF) on neuronal survival, growth, and differentiation, cultures of dissociated neonatal rat sympathetic neurons virtually free of other cell types were maintained for 3-4 wk. In the absence of NGF, the neurons did not survive for more than a day. Increased levels of NGF increased neuronal survival and growth (total protein and total lipid phosphate); saturation occurred at 0.5 microgram/ml 7S NGF. Neuronal differentiation examined by measuring catecholamine (CA) production from tyrosine also depended on the level of NGF in the culture medium. As the NGF concentration was raised, CA production per neuron, per nanogram protein, or per picomole lipid phosphate increased until saturation was achieved between 1 and 5 microgram/ml 7S NGF. Thus, NGF induces neuronal survival, growth, and differentiation of CA production in a dose-dependent fashion. Neuronal growth and differentiation were quantitatively compared in the presence of the high and low molecular weight forms of NGF; no significant functional differences were found.


[Chronic abdominal complaints and perceptions].
A recent study by Weijenborg et al. on the progress of patients with abdominal complaints actually replicates in a gynaecological outpatient clinic an earlier study carried out in an internal medicine outpatient clinic. Both studies show that only 9% of the patients are symptom-free at follow up. Perceptions of the complaints seemed related to improvement. What can a physician do for these patients? It is known from other publications that correct perception by the physician of patients' somatic attributions and anxiety leads to a decrease at the next consultation. Reduced somatic attributions and anxiety are also favourable for the outcome at six months follow up: patients have less severe abdominal pain and less healthcare-seeking behaviour. The Weijenborg et al. study is therefore a useful repeat study and reiterates that physicians' behaviour can be helpful during consultations.


Dissociation of tubular cell detachment and tubular cell death in clinical and experimental "acute tubular necrosis".
"Acute tubular necrosis" in humans is often characterized histologically by tubular cell loss and nonreplacement rather than by frank cellular necrosis. It has been assumed that tubular cells detach from the tubular basement membrane after the occurrence of irreversible cell injury and/or cell death. We examined voided urine from patients with native kidney and transplant kidney "acute tubular necrosis" and found significant tubular cell shedding with up to 100% viability of voided tubular cells, as determined by exclusion of trypan blue. Histochemical stains for the brush border enzyme gamma-glutamyl transpeptidase and electron microscopy of cell isolates demonstrated that 30 to 100% of the voided cells were of proximal tubule origin. Moreover, epithelial cell cultures were readily established from noncontaminated voided specimens, and more than 80% of the cells in monolayers expressed gamma-glutamyl transpeptidase activity in vitro. Electron microscopy of cell monolayers confirmed proximal tubular differentiation in the majority of cells in the monolayer, with the remainder having an appearance more consistent with distal origin. In rabbit models of acute ischemic (pedicle clamp) and toxic (HgCl2 and glycerol-induced) injury, tubular cell shedding was also significant, with a mean of 25 to 30% of the voided cells viable. Epithelial cells were successfully cultured and passed from animals with ischemic renal injury. In vitro studies were carried out in cultured human proximal tubular cells to assess suitability of this system for more direct study of the phenomenon of cell detachment in response to injury. We found significant changes in cell shape in response to exposure to anoxia and HgCl2 in these cultures, with extensive cell rounding and retraction at times when cell viability remained high. This in vitro system could prove useful for defining mechanisms of renal tubular cell detachment in response to injury and for examining potential interventions to prevent epithelial disruption and loss of functional integrity. These strategies may ultimately have clinical relevance in patients with "acute tubular necrosis".


Nonparaxial rogue waves in optical Kerr media.
We consider the inhomogeneous nonparaxial nonlinear Schrödinger (NLS) equation with varying dispersion, nonlinearity, and nonparaxiality coefficients, which governs the nonlinear wave propagation in an inhomogeneous optical fiber system. We present the similarity and Darboux transformations and for the chosen specific set of parameters and free functions, the first- and second-order rational solutions of the nonparaxial NLS equation are generated. In particular, the features of rogue waves throughout polynomial and Jacobian elliptic functions are analyzed, showing the nonparaxial effects. It is shown that the nonparaxiality increases the intensity of rogue waves by increasing the length and reducing the width simultaneously, by the way it increases their speed and penalizes interactions between them. These properties and the characteristic controllability of the nonparaxial rogue waves may give another opportunity to perform experimental realizations and potential applications in optical fibers.


Metagenomic Analysis of Plant Virus Occurrence in Common Bean (Phaseolus vulgaris) in Central Kenya.
Two closely related potyviruses, bean common mosaic virus (BCMV) and bean common mosaic necrosis virus (BCMNV), are regarded as major constraints on production of common bean (Phaseolus vulgaris L.) in Eastern and Central Africa, where this crop provides a high proportion of dietary protein as well as other nutritional, agronomic, and economic benefits. Previous studies using antibody-based assays and indicator plants indicated that BCMV and BCMNV are both prevalent in bean fields in the region but these approaches cannot distinguish between these potyviruses or detect other viruses that may threaten the crop. In this study, we utilized next generation shotgun sequencing for a metagenomic examination of viruses present in bean plants growing at two locations in Kenya: the University of Nairobi Research Farm in Nairobi's Kabete district and at sites in Kirinyaga County. RNA was extracted from leaves of bean plants exhibiting apparent viral symptoms and sequenced on the Illumina MiSeq platform. We detected BCMNV, cucumber mosaic virus (CMV), and Phaseolus vulgaris alphaendornaviruses 1 and 2 (PvEV1 and 2), with CMV present in the Kirinyaga samples. The CMV strain detected in this study was most closely related to Asian strains, which suggests that it may be a recent introduction to the region. Surprisingly, and in contrast to previous surveys, BCMV was not detected in plants at either location. Some plants were infected with PvEV1 and 2. The detection of PvEV1 and 2 suggests these seed transmitted viruses may be more prevalent in Eastern African bean germplasm than previously thought.


Treatment of Generalized Pustular Psoriasis of Pregnancy With Infliximab.
Generalized pustular psoriasis of pregnancy (GPPP) is a rare and severe condition that may impair the health of the mother and fetus. Effective treatment is essential, as treatment options for GPPP are limited due to concerns about unfavorable pregnancy outcomes. We report the case of a 22-year-old woman with GPPP that was unresponsive to systemic corticosteroids. We effectively treated the condition with infliximab at 30 weeks' gestation with an immediate clinical response and no detected serious adverse events except for an oral herpes infection in the patient and low birth weight in the neonate. Future studies are necessary to evaluate the safety and efficacy of infliximab treatment in GPPP.


A randomized, controlled trial of financial incentives for smoking cessation.
Smoking is the leading preventable cause of premature death in the United States. Previous studies of financial incentives for smoking cessation in work settings have not shown that such incentives have significant effects on cessation rates, but these studies have had limited power, and the incentives used may have been insufficient. We randomly assigned 878 employees of a multinational company based in the United States to receive information about smoking-cessation programs (442 employees) or to receive information about programs plus financial incentives (436 employees). The financial incentives were $100 for completion of a smoking-cessation program, $250 for cessation of smoking within 6 months after study enrollment, as confirmed by a biochemical test, and $400 for abstinence for an additional 6 months after the initial cessation, as confirmed by a biochemical test. Individual participants were stratified according to work site, heavy or nonheavy smoking, and income. The primary end point was smoking cessation 9 or 12 months after enrollment, depending on whether initial cessation was reported at 3 or 6 months. Secondary end points were smoking cessation within the first 6 months after enrollment and rates of participation in and completion of smoking-cessation programs. The incentive group had significantly higher rates of smoking cessation than did the information-only group 9 or 12 months after enrollment (14.7% vs. 5.0%, P<0.001) and 15 or 18 months after enrollment (9.4% vs. 3.6%, P<0.001). Incentive-group participants also had significantly higher rates of enrollment in a smoking-cessation program (15.4% vs. 5.4%, P<0.001), completion of a smoking-cessation program (10.8% vs. 2.5%, P<0.001), and smoking cessation within the first 6 months after enrollment (20.9% vs. 11.8%, P<0.001). In this study of employees of one large company, financial incentives for smoking cessation significantly increased the rates of smoking cessation. (ClinicalTrials.gov number, NCT00128375.)


On edge detection of x-ray images using fuzzy sets.
The effectiveness of the theory of fuzzy sets in detecting different regional boundaries of X-ray images is demonstrated. The algorithm includes a prior enhancement of the contrast among the regions (having small change in gray levels) using the contrast intensification (INT) operation along with smoothing in the fuzzy property plane before detecting its edges. The property plane is extracted from the spatial domain using S, ¿ and (1 ¿) functions and the fuzzifiers. Final edge detection is achieved using max or min operator. The system performance for different parameter conditions is illustrated by application to an image of a radiograph of the wrist.


[The immunity of patients with keratoconus].
Cellular and humoral immunity parameters were studied in patients with keratoconus at different stages, aged 14 to 49 years. The results showed the immune homeostasis to be impaired in patients with local involvement of the cornea. At a total system's level the patients developed shifts in the relative content of T-lymphocytes and their subpopulations, dysimmunoglubulinemia with increased levels of immunoglobulins of the primary immune response in the blood serum, high levels of the C3 and C4 components with a reduction of the total hemolytic activity of the complement, increased levels of NBT-positive and phagocytic cells, and a decrease of the activity of serum lysozyme.


In vivo lens deficiency of the R49C alphaA-crystallin mutant.
The R49C mutation of alphaA-crystallin (alphaA-R49C) causes hereditary cataracts in humans; patients in a four-generation Caucasian family were found be heterozygous for this autosomal dominant mutation. We previously generated knock-in mouse models of this mutation and found that by 2 months of age, heterozygous mutant mice exhibited minor lens defects including reduced protein solubility, altered signaling in epithelial and fiber cells, and aberrant interactions between alphaA-crystallin and other lens proteins. In contrast, homozygous mutant alphaA-R49C knock-in mice displayed earlier and more extensive lens defects including small eyes and small lenses at birth, death of epithelial and fiber cells, and the formation of posterior, nuclear, and cortical cataracts in the first month of life. We have extended this study to now show that in alphaA-R49C homozygous mutant mice, epithelial cells failed to form normal equatorial bow regions and fiber cells continued to die as the mice aged, resulting in a complete loss of lenses and overall eye structure in mice older than 4 months. These results demonstrate that expression of the hereditary R49C mutant of alphaA-crystallin in vivo is sufficient to adversely affect lens growth, lens cell morphology, and eye function. The death of fiber cells caused by this mutation may ultimately lead to loss of retinal integrity and blindness.


Physical layer security in light-fidelity systems.
Light-fidelity (LiFi) is a light-based wireless communication technology which can complement radio-frequency (RF) communication technologies for indoor applications. Although LiFi signals are spatially more contained than RF signals, the broadcasting nature of LiFi also makes it susceptible to eavesdropping. Therefore, it is important to secure the transmitted data against potential eavesdroppers. In this paper, an overview of the recent developments pertaining to LiFi physical layer security (PLS) is provided, and the main differences between LiFi PLS and RF PLS are explained. LiFi achievable secrecy rates and upper bounds are then investigated under practical channel models and transmission schemes. Beamforming and jamming, which received significant research attention recently as a means to achieve PLS in LiFi, are also investigated under indoor illumination constraints. Finally, future research directions of interest in LiFi PLS are identified and discussed. This article is part of the theme issue 'Optical wireless communication'.


A near-chromosome-scale genome assembly of the gemsbok (Oryx gazella): an iconic antelope of the Kalahari desert.
The gemsbok (Oryx gazella) is one of the largest antelopes in Africa. Gemsbok are heterothermic and thus highly adapted to live in the desert, changing their feeding behavior when faced with extreme drought and heat. A high-quality genome sequence of this species will assist efforts to elucidate these and other important traits of gemsbok and facilitate research on conservation efforts. Using 180 Gbp of Illumina paired-end and mate-pair reads, a 2.9 Gbp assembly with scaffold N50 of 1.48 Mbp was generated using SOAPdenovo. Scaffolds were extended using Chicago library sequencing, which yielded an additional 114.7 Gbp of DNA sequence. The HiRise assembly using SOAPdenovo + Chicago library sequencing produced a scaffold N50 of 47 Mbp and a final genome size of 2.9 Gbp, representing 90.6% of the estimated genome size and including 93.2% of expected genes according to Benchmarking Universal Single-Copy Orthologs analysis. The Reference-Assisted Chromosome Assembly tool was used to generate a final set of 47 predicted chromosome fragments with N50 of 86.25 Mbp and containing 93.8% of expected genes. A total of 23,125 protein-coding genes and 1.14 Gbp of repetitive sequences were annotated using de novo and homology-based predictions. Our results provide the first high-quality, chromosome-scale genome sequence assembly for gemsbok, which will be a valuable resource for studying adaptive evolution of this species and other ruminants.


Emerging Signal Regulating Potential of Genistein Against Alzheimer's Disease: A Promising Molecule of Interest.
Alzheimer's disease (AD) is a progressive, irreversible brain disorder characterized by pathological aggregation of the amyloid-β peptide (Aβ) and tau protein; both of these are toxic to neurons. Currently, natural products are regarded as an alternative approach to discover novel multipotent drugs against AD. Dietary soy isoflavone genistein is one of the examples of such agents that occurs naturally and is known to exert a number of beneficial health effects. It has been observed that genistein has the capacity to improve the impairments triggered by Aβ and also it possesses the antioxidant potential to scavenge the AD-mediated generation of free radicals. Furthermore, genistein can interact directly with the targeted signaling proteins and also can stabilize their activity to combat AD. In order to advance the development of AD treatment, a better comprehension of the direct interactions of target proteins and genistein might prove beneficial. Therefore, this article focuses on the therapeutic effects and molecular targets of genistein, which has been found to target directly the Aβ and tau to control the intracellular signaling pathways responsible for neurons death in the AD brain.


Immunohistochemical demonstration of the gamma2 chain of laminin-5 in urinary bladder urothelial carcinoma. Impact for diagnosis and prognosis.
The heterotrimeric molecule laminin-5 (Ln-5) represents a main protein of the epithelial adhesions complex. It links the basement membrane (BM) with the hemidesmosomes of the basal urothelial cells. The study was aimed to evaluate invasion associated changes of the epithelial adhesion complex in urothelial carcinoma (UC) monitored by immunohistochemical demonstration of the Ln-5 gamma2 chain. For correlation to UC phenotype and patients outcome, a semiquantitative immunohistochemical analysis of 100 routinely processed paraffin embedded samples (non-invasive and invasive UC) using the antibody D4B5 specific for the Ln-5 gamma2 chain was performed. An increased risk of death is associated with an increased Ln-5 loss from BM (P=0.001), an increase of stroma deposition (P=0.001), as well as an increase of cellular retention of Ln-5 protein (P=0.001) (Kruskal-Wallis test). As shown in multivariate analysis, in addition to tumor stage the cellular retention of Ln-5 is the most important prognostic parameter. In consequence, the modulation of Ln-5 is recommended as a diagnostic marker of invasive UC phenotype.


Tick-Borne Encephalitis Virus Infection Alters the Sialome of Ixodes ricinus Ticks During the Earliest Stages of Feeding.
Ticks are hematophagous arthropods that transmit a number of pathogens while feeding. Among these is tick-borne encephalitis virus (TBEV), a flavivirus transmitted by Ixodes ricinus ticks in the temperate zone of Europe. The infection results in febrile illness progressing to encephalitis and meningitis with a possibility of fatality or long-term neurological sequelae. The composition of tick saliva plays an essential role in the initial virus transmission during tick feeding. Ticks secrete a diverse range of salivary proteins to modulate the host response, such as lipocalins to control the itch and inflammatory response, and both proteases and protease inhibitors to prevent blood coagulation. Here, the effect of viral infection of adult females of Ixodes ricinus was studied with the goal of determining how the virus alters the tick sialome to modulate host tissue response at the site of infection. Uninfected ticks or those infected with TBEV were fed on mice and removed and dissected one- and 3-h post-attachment. RNA from the salivary glands of these ticks, as well as from unfed ticks, was extracted and subjected to next-generation sequencing to determine the expression of key secreted proteins at each timepoint. Genes showing statistically significant up- or down-regulation between infected and control ticks were selected and compared to published literature to ascertain their function. From this, the effect of tick viral infection on the modulation of the tick-host interface was determined. Infected ticks were found to differentially express a number of uncategorized genes, proteases, Kunitz-type serine protease inhibitors, cytotoxins, and lipocalins at different timepoints. These virus-induced changes to the tick sialome may play a significant role in facilitating virus transmission during the early stages of tick feeding.


[The African and Malagasy Council for higher education: objectives and programs].
The African and Malgach Council for University Education is an inter-african institution. The anthors remind its organisation and show its aim and basic programms: recognition and equivalence of the degrees--teaching of the traditional medication--establishment of inter-african comettees--organisation of concors for associated professors in laws and medicine.


[Sufficiency of income perceived by the elderly in the city of São Paulo/Brazil].
During the course of their lives elderly Brazilians have witnessed a country with major social contradictions, where the differences between individuals in relation to the factors involved in treatment and understanding of their needs are many. In 2000, factors associated with the perception of income sufficiency of elderly people (60 years and over) in São Paulo were analyzed, in an attempt to understand a few of these aspects. A cross-sectional epidemiological study was conducted by interviews with 2,143 elderly people (60 years and over). Of these, 1,300 needed no help in answering the questionnaire and scored 13 or more points on the Mini Mental State Examination. They had salaried work at some stage, had some form of income and this matched the amount of income sources reported. Thirty-one percent of the elderly perceived their income as sufficient. Those with income above poverty levels had higher odds of satisfactory perception. Regardless of income, they were associated with: having predominantly worked in non-physical labor; started work at 14 years or older; having private health insurance; more than one source of income; not suffering privation in the first 15 years of life; being 70 or older; enjoying good health and autonomy in daily activities without assistance.


Condom availability programs in U.S. schools.
School condom availability programs have been promoted as a promising approach for increasing condom use among students, for reducing the risk of infections with the human immunodeficiency virus and with other sexually transmitted diseases and for preventing unintended pregnancy. Data from a telephone survey of key individuals at school condom programs across the United States suggest that as of January 1995, at least 431 public schools in 50 U.S. school district made condoms available-2.2% of all public high schools and 0.3% of high school districts. In about half of the schools that were surveyed, students obtained more than one condom per student per year, on average, and in 14% students obtained more than six. Students in alternative schools, in smaller schools, in schools that made condoms available in baskets and in schools with health clinics obtained more condoms per student per year than did students in other schools.


Impact of nonexacerbated COPD on mortality in critically ill patients.
The real contribution of nonexacerbated COPD to mortality has not been studied. The aim of our study was to evaluate the impact of nonexacerbated COPD on mortality in patients requiring mechanical ventilation (MV). This prospective, observational study included critically ill, ventilated patients without evidence of respiratory infection. Patients with COPD comprised the study group. Clinical and demographics variables were recorded. The main end point was ICU mortality. Of the 235 patients included, 60 (25.5%) intubated patients had COPD. The remaining 175 (74.5%) comprised the control group. Those with COPD were more often medical patients who were older and had a higher number of comorbidities and a higher APACHE (Acute Physiology and Chronic Health Evaluation) II score than intubated patients without COPD (P < .05). The overall ICU mortality was 26.3% (62/235) and significantly higher in patients with nonexacerbated COPD (36.7% vs 22.9%, P < .05), with an attributable mortality to COPD of 13.8%. Incidence of ventilator-associated pneumonia was not significantly different between patients with nonexacerbated COPD (11.9/1,000 MV days) and without COPD (16.0/1,000 MV days; P = .40). In the multivariate analysis, only COPD (hazard ratio [HR], 2.1; 95% CI, 1.10-3.94), shock at ICU admission (HR, 2.0; 95% CI, 1.01-4.01), and medical condition (HR, 1.7; 95% CI, 1.01-3.18) were independently associated with mortality. Intubated patients with nonexacerbated COPD were not exposed to a higher risk of ventilator-associated pneumonia but had a higher rate of mortality.


Patient health outcomes in psychiatric mental health nursing.
This integrative literature review examined evidence concerning the relationship between psychiatric mental health nursing interventions and patient-focused outcomes. Empirical studies, published between 1997 and 2007, were identified and gathered by searching relevant databases and specific data sources. Although 156 articles were critically appraised, only 25 of them met the inclusion criteria. Findings from this review showed that the most frequently used outcome instruments assessed psychiatric symptom severity. Most of the instruments targeted two symptom categories: altered thoughts/perceptions and altered mood. Other outcome instruments were categorized in the following domains: self-care, functioning, quality of life and satisfaction. The most important finding of this review is the lack of consistently strong evidence to support decisions concerning which outcome instrument or combination of instruments to recommend for routine use in practice. Based on this review, additional research to conceptualize, measure and examine the feasibility of outcome instruments sensitive to psychiatric mental health nursing interventions is recommended.


[Pre- and post-conditioning phenomena: the protective physiological mechanisms in the aspect of pathogenesis and the theory of treatment of ENT pathology].
The objective of the present work was to expose the universal general biological significance of the protective pre- and postconditioning phenomena and to provide an insight into the possibility of application of therapeutic modalities based on these effects in current otorhinolaryngological practice. Pre- and postconditioning phenomena (Pre-C and Post-C respectively) began to be studied as protective physiological mechanisms since the 1980s, first in cardiology and thereafter in other fields of experimental medicine. At the same time, their protective properties had been known and intuitively used much earlier among the established human cultural and social stereotypes, psychophysical training techniques, and methods of traditional and empirical medicine. The widespread application of these phenomena gives evidence of their universal biological nature as factors involved in the interactions between the organism and pathogens (including co-morbidity), the process leading to the enhancement of non-specific resistance, mechanisms underlying realization of pharmacodynamic effects of a number of pharmaceutical products,etc. The understanding of the protective potential of PreC and PostC dictates the necessity to revise and further elaborate the present-day strategy of prophylaxis and treatment of the most serious chronic ENT diseases.


Three-dose vs extended-course clindamycin prophylaxis for free-flap reconstruction of the head and neck.
Twenty-four hours of perioperative antibiotics provides effective prophylaxis for most head and neck cancer resections. Many reconstructive surgeons have been hesitant to apply this standard to free-flap reconstruction of the head and neck. This prospective clinical trial compared short-course and long-course clindamycin prophylaxis for wound infection in patients with head and neck cancer undergoing free-flap reconstruction. Seventy-four patients were randomized to receive short-course (3 doses) or long-course (15 doses) clindamycin perioperatively. Wound infections, fistulas, and other postoperative complications were documented by faculty surgeons who were blinded as to treatment group. The differences in wound infections and other complications were statistically insignificant. No other independent predictors of wound complications emerged in this series of patients. Short-course clindamycin is as effective as long-course clindamycin in preventing wound infections after free-flap surgery for head and neck ablative defects.


Numerous risk factors for Helicobacter pylori antibiotic resistance revealed by extended anamnesis: a Bulgarian study.
The aim of this study was to assess risk factors for primary Helicobacter pylori antibiotic resistance by an extended anamnesis. In total, 519 H. pylori strains from untreated symptomatic adults who answered a questionnaire were evaluated. Strain susceptibility was assessed by a breakpoint susceptibility test. Primary resistance rates were 29.5 % for metronidazole, 17.9 % for clarithromycin, 7.3 % for metronidazole+clarithromycin, 4.0 % for tetracycline and 10.8 % for ciprofloxacin. On multivariate analysis, younger (≤65 years) age was an independent predictor for metronidazole resistance. To our knowledge, for the first time, being a member of the health-care profession was revealed as a risk factor for H. pylori resistance to metronidazole and both metronidazole and clarithromycin. Respiratory and urinary tract infections were independent predictors of clarithromycin and ciprofloxacin resistance, respectively. The presence of co-infections was an independent risk factor for clarithromycin, metronidazole and ciprofloxacin resistance. Surprisingly, female sex was the only predictor for tetracycline resistance. The antibiotic resistance rates were not associated with disease type, place of residence, birthplace, educational level, non-steroidal anti-inflammatory drug or proton pump inhibitor use, smoking or dietary factors, such as consumption of coffee, yogurt, green tea, raw garlic, raw onion, honey or meat. There was a trend for higher metronidazole resistance in strains from diabetic patients. In conclusion, the extended anamnesis of H. pylori-positive patients should include data on patient age, sex, whether they are in the health-care profession, co-infections and possibly diabetes to improve the choice of empiric therapy. Tailored treatment based on the extended anamnesis is suggested, and susceptibility testing of the strains is recommended for patients at risk for antibiotic resistance, especially to clarithromycin, fluoroquinolones or both metronidazole and clarithromycin.


What's the Purpose of Rounds? A Qualitative Study Examining the Perceptions of Faculty and Students.
Rounds are a critical activity on any inpatient service, but there is little literature describing the purpose of rounds from the perspective of faculty and trainees in teaching hospitals. To evaluate and compare the perceptions of pediatric and internal medicine attendings and medical students regarding the purpose of inpatient attending rounds. The authors conducted 10 semistructured focus groups with attendings and medical students in the spring of 2014 at 4 teaching hospitals. The protocol was approved by the institutional review boards at all institutions. The authors employed a grounded theory approach to data collection and analysis, and data were analyzed by using the constant- comparative method. Two transcripts were read and coded independently by 2 authors to generate themes. Forty-eight attendings and 31 medical students participated in the focus groups. We categorized 218 comments into 4 themes comprised of 16 codes representing what attendings and medical students believed to be the purpose of rounds. These themes included communication, medical education, patient care, and assessment. Our results highlight that rounds serve 4 purposes, including communication, medical education, patient care, and assessment. Importantly, both attendings and students agree on what they perceive to be the many purposes of rounds. Despite this, a disconnect appears to exist between what people believe are the purposes of rounds and what is happening during rounds.


Differential regulation of antioxidant enzymes in Frankliniella occidentalis (Thysanoptera: Thripidae) exposed to thermal stress.
Frankliniella occidentalis is an invasive insect pest that incites damage to ornamental and agronomic crops on a global scale. In this study, the effects of temperature on gene expression and enzyme activity were studied for superoxide dismutase (SOD), peroxidase (POD), and glutathione-S-transferase (GST) in F. occidentalis. SOD, POD and GST enzyme activity increased significantly at 35-37 °C but declined as the temperature increased to 41 °C. In a time course study at 35 °C, SOD, POD and GST activities were significantly elevated at 0.5, 1 and 2 h in comparison to the control at 26 °C. Expression patterns were evaluated for the three antioxidant genes under high and low temperature stress. In a time course study at -4 °C, SOD, POD and GST expression peaked at 1 h and declined at 2 h of exposure. In contrast, when transcription was monitored at 35 °C, expression was lowest at 1 h and increased at 2 h. The results provide data that will be useful in deciphering the role of antioxidant enzymes in the adaptation of F. occidentalis to climate change.


What do we know about community-based health worker programs? A systematic review of existing reviews on community health workers.
To synthesize current understanding of how community-based health worker (CHW) programs can best be designed and operated in health systems. We searched 11 databases for review articles published between 1 January 2005 and 15 June 2017. Review articles on CHWs, defined as non-professional paid or volunteer health workers based in communities, with less than 2 years of training, were included. We assessed the methodological quality of the reviews according to AMSTAR criteria, and we report our findings based on PRISMA standards. We identified 122 reviews (75 systematic reviews, of which 34 are meta-analyses, and 47 non-systematic reviews). Eighty-three of the included reviews were from low- and middle-income countries, 29 were from high-income countries, and 10 were global. CHW programs included in these reviews are diverse in interventions provided, selection and training of CHWs, supervision, remuneration, and integration into the health system. Features that enable positive CHW program outcomes include community embeddedness (whereby community members have a sense of ownership of the program and positive relationships with the CHW), supportive supervision, continuous education, and adequate logistical support and supplies. Effective integration of CHW programs into health systems can bolster program sustainability and credibility, clarify CHW roles, and foster collaboration between CHWs and higher-level health system actors. We found gaps in the review evidence, including on the rights and needs of CHWs, on effective approaches to training and supervision, on CHWs as community change agents, and on the influence of health system decentralization, social accountability, and governance. Evidence concerning CHW program effectiveness can help policymakers identify a range of options to consider. However, this evidence needs to be contextualized and adapted in different contexts to inform policy and practice. Advancing the evidence base with context-specific elements will be vital to helping these programs achieve their full potential.


The unit in preference to the titre as a measure of agglutinating activity.
The agglutinating activity of individual sera is usually expressed by a "titre" notation, the titre being dependent on the degree of agglutination arbitrarily chosen as the end-point. This titre may be corrected by comparison with a standard serum. A "unit" notation, however, necessarily requires both comparative measurements in relation to a standard serum and the taking into consideration of all degrees of agglutination. This involves log-dose-response curves for the standard serum and the test serum; and cases in which the distance between such curves is not constant, either at different degrees of agglutination or where different antigen suspensions are used, will bring to light fundamental limitations of the bio-assay which the simple use of titres may not reveal.


Impaired autophagic flux is critically involved in drug-induced dopaminergic neuronal death.
Autophagy is an evolutionarily conserved process that mediates the degradation of abnormal proteins and the removal of dysfunctional organelles. Recently, accumulating evidence has implicated the dysregulation of autophagy as underlying the pathophysiology of several neurodegenerative diseases. Using culture models of Parkinson's disease, we have investigated whether and how prototypic autophagic events occur upon exposure to N-methyl-4-phenylpyridinium, a dopaminergic neurotoxin, or nigericin, a K(+)/H(+) ionophore. From these independent studies, we have found that these drugs equally induce morphological and biochemical changes typical of autophagy, including accumulation of autophagic vacuoles, appearance of LC3-II forms, and alteration in the expression and distribution of p62. Further investigation has indicated that drug-induced autophagic phenomena are largely the consequences of an impaired autophagic flux. In these cell death paradigms, we have intriguingly found that Bak, a prototypic proapoptotic protein of the Bcl-2 family, exerts a protective role via reduction of the area occupied by swollen vacuoles and appearance of the LC3-II form, whereas silencing of Bak aggravates these phenomena. Further study has indicated that a protective role for Bak is primarily ascribed to its regulatory effect on the maintenance of autophagic flux and vacuole homeostasis. In this regard, a regulatory role for calcium has been proposed.


Probability-Based Algorithm for Bearing Diagnosis with Untrained Spall Sizes.
Bearing spall detection and predicting its size are great challenges. Model-based simulation is a well-known traditional approach to physically model the influence of the spall on the bearing. Building a physical model is challenging due to the bearing complexity and the expert knowledge required to build such a model. Obviously, building a partial physical model for some of the spall sizes is easier. In this paper, we propose a machine-learning algorithm, called Probability-Based Forest, that uses a partial physical model. First, the behavior of some of the spall sizes is physically modeled and a simulator based on this model generates scenarios for these spall sizes in different conditions. Then, the machine-learning algorithm trains these scenarios to generate a prediction model of spall sizes even for those that have not been modeled by the physical model. Feature extraction is a key factor in the success of this approach. We extract features using two traditional approaches: statistical and physical, and an additional new approach: Time Series FeatuRe Extraction based on Scalable Hypothesis tests (TSFRESH). Experimental evaluation with well-known physical model shows that our approach achieves high accuracy, even in cases that have not been modeled by the physical model. Also, we show that the TSFRESH feature-extraction approach achieves the highest accuracy.


Rotamer Libraries for the High-Resolution Design of β-Amino Acid Foldamers.
β-Amino acids offer attractive opportunities to develop biologically active peptidomimetics, either employed alone or in conjunction with natural α-amino acids. Owing to their potential for unique conformational preferences that deviate considerably from α-peptide geometries, β-amino acids greatly expand the possible chemistries and physical properties available to polyamide foldamers. Complete in silico support for designing new molecules incorporating non-natural amino acids typically requires representing their side-chain conformations as sets of discrete rotamers for model refinement and sequence optimization. Such rotamer libraries are key components of several state-of-the-art design frameworks. Here we report the development, incorporation in to the Rosetta macromolecular modeling suite, and validation of rotamer libraries for β3-amino acids.


Tau-based fluorescent protein fusions to visualize microtubules.
The ability to visualize cytoskeletal proteins and their dynamics in living cells has been critically important in advancing our understanding of numerous cellular processes, including actin- and microtubule (MT)-dependent phenomena such as cell motility, cell division, and mitosis. Here, we describe a novel set of fluorescent protein (FP) fusions designed specifically to visualize MTs in living systems using fluorescence microscopy. Each fusion contains a FP module linked in frame to a modified phospho-deficient version of the MT-binding domain of Tau (mTMBD). We found that expressed and purified constructs containing a single mTMBD decorated Xenopus egg extract spindles more homogenously than similar constructs containing the MT-binding domain of Ensconsin, suggesting that the binding affinity of mTMBD is minimally affected by localized signaling gradients generated during mitosis. Furthermore, MT dynamics were not grossly perturbed by the presence of Tau-based FP fusions. Interestingly, the addition of a second mTMBD to the opposite terminus of our construct caused dramatic changes to the spatial localization of probes within spindles. These results support the use of Tau-based FP fusions as minimally perturbing tools to accurately visualize MTs in living systems.


The comparative study of metacognition: sharper paradigms, safer inferences.
Results that point to animals' metacognitive capacity bear a heavy burden, given the potential for competing behavioral descriptions. In this article, formal models are used to evaluate the force of these descriptions. One example is that many existing studies have directly rewarded so-called uncertainty responses. Modeling confirms that this practice is an interpretative danger because it supports associative processes and encourages simpler interpretations. Another example is that existing studies raise the concern that animals avoid difficult stimuli not because of uncertainty monitored, but because of aversion given error-causing or reinforcement-lean stimuli. Modeling also justifies this concern and shows that this problem is not addressed by the common practice of comparing performance on chosen and forced trials. The models and related discussion have utility for metacognition researchers and theorists broadly, because they specify the experimental operations that will best indicate a metacognitive capacity in humans or animals by eliminating alternative behavioral accounts.


Muscle biopsy and cell cultures: potential diagnostic tools in hereditary skeletal muscle channelopathies.
Hereditary muscle channelopathies are caused by dominant mutations in the genes encoding for subunits of muscle voltage-gated ion channels. Point mutations on the human skeletal muscle Na+ channel (Nav1.4) give rise to hyperkalemic periodic paralysis, potassium aggravated myotonia, paramyotonia congenita and hypokalemic periodic paralysis type 2. Point mutations on the human skeletal muscle Ca2+ channel give rise to hypokalemic periodic paralysis and malignant hyperthermia. Point mutations in the human skeletal chloride channel CIC-1 give rise to myotonia congenita. Point mutations in the inwardly rectifying K+ channel Kir2.1 give rise to a syndrome characterized by periodic paralysis, severe cardiac arrhythmias and skeletal alterations (Andersen's syndrome). Involvement of the same ion channel can thus give rise to different phenotypes. In addition, the same mutation can lead to different phenotypes or similar phenotypes can be caused by different mutations on the same or on different channel subtypes. Bearing in mind, the complexity of this field, the growing number of potential channelopathies (such as the myotonic dystrophies), and the time and cost of the genetic procedures, before a biomolecular approach is addressed, it is mandatory to apply strict diagnostic protocols to screen the patients. In this study we propose a protocol to be applied in the diagnosis of the hereditary muscle channelopathies and we demonstrate that muscle biopsy studies and muscle cell cultures may significantly contribute towards the correct diagnosis of the channel involved. DNA-based diagnosis is now a reality for many of the channelopathies. This has obvious genetic counselling, prognostic and therapeutic implications.


Inhibition of p56(lck) tyrosine kinase by isothiazolones.
Lck encodes a 56-kDa protein-tyrosine kinase, predominantly expressed in T lymphocytes, crucial for initiating T cell antigen receptor (TCR) signal transduction pathways, culminating in T cell cytokine gene expression and effector functions. As a consequence of a high-throughput screen for selective, novel inhibitors of p56(lck), an isothiazolone compound was identified, methyl-3-(N-isothiazolone)-2-thiophenecarboxylate(A-125800), which inhibits p56(lck) kinase activity with IC50 = 1-7 microM. Under similar assay conditions, the isothiazolone compound was equipotent in blocking the ZAP-70 tyrosine kinase activity but was 50 to 100 times less potent against the catalytic activities of p38 MAP kinase and c-Jun N-terminal kinase 2alpha. A-125800 blocked activation-dependent TCR tyrosine phosphorylation and intracellular calcium mobilization in Jurkat T cells (IC50 = 35 microM) and blocked T cell proliferation in response to alloantigen (IC50 = 14 microM) and CD3/CD28-induced IL-2 secretion (IC50 = 2.2 microM) in primary T cell cultures. Inhibition of p56(lck )by A-125800 was dose- and time-dependent and was irreversible. A substitution of methylene for the sulfur atom in the isothiazolone ring of the compound completely abrogated the ability to inhibit p56(lck) kinase activity and TCR-dependent signal transduction. Incubation with thiols such as beta-ME or DTT also blocked the ability of the isothiazolone to inhibit p56(lck) kinase activity. LC/MS analysis established the covalent modification of p56(lck) at cysteine residues 378, 465, and 476. Together these data support an inhibitory mechanism, whereby cysteine -SH groups within the p56(lck) catalytic domain react with the isothiazolone ring, leading to ring opening and disulfide bond formation with the p56(lck) enzyme. Loss of p56(lck) activity due to -SH oxidation has been suggested to play a role in the pathology of AIDS. Consequently, a similar mechanism of sulfhydryl oxidation leading to p56(lck) inhibition, described in this report, may occur in the intact T cell and may underlie certain T cell pathologies.


Illuminating COI1: a component of the Arabidopsis jasomonate receptor complex also interacts with ethylene signaling.
A significant portion of developmental and environmental responses in plants is mediated through phytohormone signaling, often if not always integrated with outputs from other signals. We have recently shown that CORONATINE INSENSITIVE1 (COI1), a component of a jasmonate receptor complex, is involved in ethylene-induced root growth inhibition of Arabidopsis, in the light. This response is neither due to elevated levels of jasmonates in response to ethylene treatment nor dependent on the known jasmonate signal-transduction cascade, except that it requires COI1. Further, we have shown that the ethylene-induced COI1-mediated pathway functions in parallel with, and additively to, the conventional ethylene signaling pathway, and that the light requirement is primarily for long photoperiods. This unexpected interaction of COI1 with ethylene signaling has also been extended to other developmental processes including germination and fertility. This addendum summarizes the earlier findings with some new insights, and describes and speculates on the mechanisms by which these processes are regulated, in the context of the interaction between COI1 and ethylene signaling.