Therapy with mud compresses for knee osteoarthritis: comparison of natural mud preparations with mineral-depleted mud.
Mud pack therapy is an alternative mode of treatment for rheumatic diseases. It is based on the application of heated mud packs to the entire body or to specific areas, such as over joints. The aim of the current study was to evaluate the efficacy of treatment with mud compresses at patients' homes for osteoarthritis of the knee. Fifty-eight patients with osteoarthritis of the knee were enrolled in a prospective, double-blinded, controlled study. Forty patients were treated with natural mineral-rich mud compresses and 18 patients were treated with mineral-depleted mud compresses. Mud compresses were applied 5 times each week during 3 weeks for a total of 15 treatments. Patients were assessed at baseline, at completion of the 3-week treatment period, and twice after the conclusion of the treatment period-after 1 month and after 3 months. The main outcome measures were the Lequesne Index of severity of knee osteoarthritis, patient self-assessment of pain, and severity of knee pain on a visual analog scale. A reduction of 20% or more in the pain scores was considered clinically significant. In the group treated with natural mud compresses, a significant reduction in knee pain was observed at all assessments. Similarly, improvement in the Lequesne Index was seen at the end of therapy and a month after treatment. In the control group, given mineral-depleted mud compresses, no significant change in knee pain was seen at any assessment. Improvement in the Lequesne Index was seen 1 and 3 months after completion of the therapy, but not at the end of therapy. Seventy-two percent of the patients in the treatment group had an improvement of >20% in self-assessment of knee pain, compared with 33% in the control group (p = 0.005). The data suggest that treatment with mud compresses, but only in their natural form, temporarily relieves pain in patients with osteoarthritis of the knees. We believe that treatment with mud compresses might augment conventional medical therapy in these patients.


Long-term carbimazole pretreatment reduces the efficacy of radioiodine therapy.
Data from several studies suggest that pretreatment with antithyroid drugs (ATD) before (131)I increases the risk of treatment failure. This effect has been demonstrated more consistently with propylthiouracil than with carbimazole (CMZ) or methimazole (MMI). Men with Graves' disease (GD) have a lower rate of remission with (131)I compared to women and the impact of long-term ATD pretreatment on the success of (131)I is unknown. The objective of our study was to compare the efficacy of fixed doses of radioiodine between patients with and without long-term CMZ pretreatment. We performed a retrospective study on 335 male patients with GD treated with (131)I from 1998 to 2008. 148 patients had been pretreated with CMZ, and the remaining 187 patients received (131)I without pretreatment. We compared the success rate of a single dose of (131)I, between patients with and without long-term CMZ pretreatment. The success rate of a single dose of (131)I was significantly higher in patients without pretreatment than in patients who were pretreated with CMZ (91.4% vs. 82.3%, P = 0.01). The rate of hypothyroidism in the first 6 months after (131)I therapy was significantly higher in patients without pretreatment (55.1% vs. 44.6%, P = 0.05). There was also a trend for higher cumulative rate of hypothyroidism at last follow-up in nonpretreated patients (78.1% vs. 69.7%). Male patients with Graves' hyperthyroidism pretreated with CMZ have lower efficacy with 131I therapy compared to nonpretreated patients. CMZ pretreatment given for a prolonged period reduces the efficacy of (131)I therapy.


Quantitative trait loci mapping in dairy cattle: review and meta-analysis.
From an extensive review of public domain information on dairy cattle quantitative trait loci (QTL), we have prepared a draft online QTL map for dairy production traits. Most publications (45 out of 55 reviewed) reported QTL for the major milk production traits (milk, fat and protein yield, and fat and protein concentration (%)) and somatic cell score. Relatively few QTL studies have been reported for more complex traits such as mastitis, fertility and health. The collated QTL map shows some chromosomal regions with a high density of QTL, as well as a substantial number of QTL at single chromosomal locations. To extract the most information from these published records, a meta-analysis was conducted to obtain consensus on QTL location and allelic substitution effect of these QTL. This required modification and development of statistical methodologies. The meta-analysis indicated a number of consensus regions, the most striking being two distinct regions affecting milk yield on chromosome 6 at 49 cM and 87 cM explaining 4.2 and 3.6 percent of the genetic variance of milk yield, respectively. The first of these regions (near marker BM143) affects five separate milk production traits (protein yield, protein percent, fat yield, fat percent, as well as milk yield).


The I1307K APC polymorphism in Ashkenazi Jews with colorectal cancer: clinical and pathologic features.
Colorectal cancer is common in Ashkenazi Jews. The I1307K APC mutation occurs in 6-7% of Ashkenazi Jews and increases the risk of colorectal cancer. This study aimed to describe the clinical, pathologic and epidemiologic features of colorectal cancer in I1307K carriers to determine whether there were any features which might warrant individual screening for the mutation. In all, 215 Ashkenazi Jews with a personal history of colorectal cancer were enrolled. Clinical and family history, pathology reports, and slides were obtained and blood drawn for I1307K determination. The presence of the mutation was determined by PCR from white blood cell DNA. Colorectal cancer pathology slides were read in a blinded fashion. Of the 215 enrolled patients, 26 (12.1%) tested positive for I1307K. There was no difference in the pathologic features between colorectal cancers in Ashkenazi carriers compared to noncarriers. There was no difference in the age at diagnosis or history of second or other primaries. Carriers had an increased likelihood of having a first-degree relative with colorectal cancer (50%) compared to noncarriers (28%, P < 0.04). We could find no distinguishing feature other than family history that characterizes I1307K positive colorectal cancers. We could find no group of Ashkenazi Jews with colorectal cancer for whom screening for I1307K would be clinically useful.


The role of MEKK1 in hypertrophic cardiomyopathy.
MEKK1 is a ubiquitously expressed mitogen activated protein kinase that is involved in tissue remodeling in a variety of settings including carotid artery blood flow cessation, wound healing, and breast adenocarcinoma intravasation. Here, we have tested the function of MEKK1 in genetic hypertrophic cardiomyopathy (HCM). MEKK1 was genetically deleted in C57Bl6/J mice expressing a mutant alpha-myosin heavy chain (HCM-MEKK1(-/-)). The absence of MEKK1 in HCM resulted in a more pronounced hypertrophy when compared to HCM mice with the MEKK1 gene intact without further increases in atrial natriuretic factor and beta-myosin heavy chain (MyHC) expression and fibrosis. Since MEKK1 is required for the induction of several tissue proteases, we tested the hypothesis that cardiac enlargement of HCM- MEKK1(-/-) mice was due to altered expression of urokinase-type plasminogen activator (uPA), JunB, matrix-metalloproteinase (MMP), and tissue inhibitors of MMPs (TIMPs). Because of its role in preventing apoptosis, we also tested the loss of MEKK1 on apoptotic mediators Bcl-2, cytochrome C, caspase-9, and caspase-3. uPA expression was decreased while JunB, MMP-9, caspase-9, and caspase-3 activities were elevated in HCM- MEKK1(-/-) hearts when compared to MEKK1(-/-), wild-type (WT), and HCM mice. Bcl-2 and Cyt C expression was elevated only in HCM mice. We conclude that the absence of MEKK1 induces a more pronounced cardiac hypertrophy to HCM through altered expression of proteases implicated in cardiac remodeling and increased apoptosis.


Warming experiments underpredict plant phenological responses to climate change.
Warming experiments are increasingly relied on to estimate plant responses to global climate change. For experiments to provide meaningful predictions of future responses, they should reflect the empirical record of responses to temperature variability and recent warming, including advances in the timing of flowering and leafing. We compared phenology (the timing of recurring life history events) in observational studies and warming experiments spanning four continents and 1,634 plant species using a common measure of temperature sensitivity (change in days per degree Celsius). We show that warming experiments underpredict advances in the timing of flowering and leafing by 8.5-fold and 4.0-fold, respectively, compared with long-term observations. For species that were common to both study types, the experimental results did not match the observational data in sign or magnitude. The observational data also showed that species that flower earliest in the spring have the highest temperature sensitivities, but this trend was not reflected in the experimental data. These significant mismatches seem to be unrelated to the study length or to the degree of manipulated warming in experiments. The discrepancy between experiments and observations, however, could arise from complex interactions among multiple drivers in the observational data, or it could arise from remediable artefacts in the experiments that result in lower irradiance and drier soils, thus dampening the phenological responses to manipulated warming. Our results introduce uncertainty into ecosystem models that are informed solely by experiments and suggest that responses to climate change that are predicted using such models should be re-evaluated.


Anesthetic Considerations for Automated High-Frequency Jet Ventilation During Electromagnetic Navigation Bronchoscopy.
Newly advanced diagnostic bronchoscopic procedures, such as electromagnetic navigation bronchoscopy using navigation system technology (superDimension, Medtronic), provides computed tomography referenced and computerized 3-dimensional imaging. To increase accuracy and higher diagnostic biopsy yield, electromagnetic navigation bronchoscopy necessitates special anesthetic and ventilation techniques providing the interventional pulmonologist minimal respiratory lung motion. This anesthetic meets 2 important goals by limiting almost all interference from diaphragmatic and lung movement while allowing the anesthesia provider to achieve hands-free management. Proposed here is an anesthetic ventilation technique by automated high-frequency jet ventilation (HFJV) via double-lumen micro jet endotracheal tubes. This ventilation technique delivers consistent very low tidal volumes. Automated HFJV provides the pulmonologist the advantage of more accurate navigation and target alignment in this Global Positioning System-guided biopsy procedure. The technique offers essentially no chest motion, without interrupting ventilation. Additionally, HFJV allows the anesthetist better availability to attend to total intravenous anesthesia, adjustments, and interventions. The intention of this article is to detail an anesthetic method that provides a hands-free technique that requires only one anesthesia provider.


Effects of prenatal nicotine exposure on development of central and peripheral cholinergic neurotransmitter systems. Evidence for cholinergic trophic influences in developing brain.
The development of cholinergic systems in brain regions was evaluated biochemically in developing control rats and rats whose mothers received nicotine via continuous minipump infusion during gestational days 4 to 20. The cerebral cortex displayed a unique maturational pattern of choline acetyltransferase (ChAT) activity and high-affinity synaptosomal [3H]choline uptake capabilities, characterized by increases in the concentration of these components and a postnatal spike of neuronal activity as assessed with the uptake/ChAT ratio; the peak of activity coincided with the time at which neurogenesis declines and synaptogenesis rises. Evaluation of the same markers in midbrain + brainstem indicated rises in uptake which were relatively unselective, primarily reflecting tissue growth and no postnatal peak of uptake/ChAT; cerebellum likewise showed primarily tissue growth-related changes in ChAT rather than increases in its specific concentration. Prenatal exposure to nicotine had a marked adverse effect on developmental patterns of ChAT, uptake and uptake/ChAT only in cerebral cortex, the region previously shown to exhibit major abnormalities caused by this drug and other treatments with cholinomimetic effects. ChAT was unaffected in peripheral projections to the adrenal. Nicotine may thus selectively disrupt central nervous system development by stimulating nicotinic receptors which are present in fetal brain, prematurely eliciting the events ordinarily triggered postnatally by cholinergic projections.


A comprehensive comparative review of sequence-based predictors of DNA- and RNA-binding residues.
Motivated by the pressing need to characterize protein-DNA and protein-RNA interactions on large scale, we review a comprehensive set of 30 computational methods for high-throughput prediction of RNA- or DNA-binding residues from protein sequences. We summarize these predictors from several significant perspectives including their design, outputs and availability. We perform empirical assessment of methods that offer web servers using a new benchmark data set characterized by a more complete annotation that includes binding residues transferred from the same or similar proteins. We show that predictors of DNA-binding (RNA-binding) residues offer relatively strong predictive performance but they are unable to properly separate DNA- from RNA-binding residues. We design and empirically assess several types of consensuses and demonstrate that machine learning (ML)-based approaches provide improved predictive performance when compared with the individual predictors of DNA-binding residues or RNA-binding residues. We also formulate and execute first-of-its-kind study that targets combined prediction of DNA- and RNA-binding residues. We design and test three types of consensuses for this prediction and conclude that this novel approach that relies on ML design provides better predictive quality than individual predictors when tested on prediction of DNA- and RNA-binding residues individually. It also substantially improves discrimination between these two types of nucleic acids. Our results suggest that development of a new generation of predictors would benefit from using training data sets that combine both RNA- and DNA-binding proteins, designing new inputs that specifically target either DNA- or RNA-binding residues and pursuing combined prediction of DNA- and RNA-binding residues.


Detection of CTX-M-15 harboring Escherichia coli isolated from wild birds in Tunisia.
The spreading of antibiotic resistant bacteria is becoming nowadays an alarming threat to human and animal health. There is increasing evidence showing that wild birds could significantly contribute to the transmission and spreading of drug-resistant bacteria. However, data for antimicrobial resistance in wild birds remain scarce, especially throughout Africa. The aims of this investigation were to analyze the prevalence of ESBL-producing E. coli in faecal samples of wild birds in Tunisia and to characterize the recovered isolates. One hundred and eleven samples were inoculated on MacConkey agar plates supplemented with cefotaxime (2 μg/ml). ESBL-producing E. coli isolates were detected in 12 of 111 faecal samples (10.81%) and one isolate per sample was further characterized. β-lactamase detected genes were as follows: blaCTX-M-15 (8 isolates), blaCTX-M-15 + blaTEM-1b (4 isolates). The ISEcp1 and orf477 sequences were found respectively in the regions upstream and downstream of all blaCTX-M-15 genes. Seven different plasmid profiles were observed among the isolates. IncF (FII, FIA, FIB) and IncW replicons were identified in 11 CTX-M-15 producing isolates, and mostly, other replicons were also identified: IncHI2, IncA/C, IncP, IncI1 and IncX. All ESBL-producing E. coli isolates were integron positive and possessed "empty" integron structures with no inserted region of DNA. The following detected virulence genes were: (number of isolates in parentheses): fimA (ten); papC (seven); aer (five); eae (one); and papGIII, hly, cnf, and bfp (none). Molecular typing using pulsed-field gel electrophoresis and multilocus sequence typing showed a low genetic heterogeneity among the 12 ESBL-producing strains with five unrelated PFGE types and five different sequence types (STs) respectively. CTX-M-15-producing isolates were ascribed to phylogroup A (eleven isolates) and B2 (one isolate). To our knowledge, this study provides the first insight into the contribution of wild birds to the dynamics of ESBL-producing E. coli in Tunisia.


Association between mid- to late life physical fitness and dementia: evidence from the CAIDE study.
This study investigated the association between perceived physical fitness at midlife, changes in perceived fitness during the three decades from mid- to late life and dementia risk. Prospective cohort study. Cardiovascular risk factors, ageing and incidence of dementia (CAIDE) study. Subjects were selected from four independent, random samples of population-based cardiovascular surveys and were first examined in 1972, 1977, 1982 or 1987, when they were on average 50 years old. The CAIDE target population included 3559 individuals. A random sample of 2000 individuals still alive in 1997 was drawn for re-examinations (performed in 1998 and 2005-2008) that consisted of cognitive assessments, with 1511 subjects participating in at least one re-examination. Dementia diagnoses were also confirmed from national registers for the entire target population. All-cause dementia. Poor physical fitness at midlife was associated with increased dementia risk in the entire target population [hazard ratio (HR), 1.5; 95% confidence interval (CI), 1.1-2.0]. In participants, odds ratio (OR) was 2.0 (95% CI, 0.9-4.0). This association was significant in apolipoprotein E ε4 allele (APOEε4) noncarriers (OR, 4.3; 95% CI, 1.4-13.3), men (HR, 1.8; 95% CI, 1.1-3.0) and people with chronic conditions (HR, 2.9; 95% CI, 1.3-6.6). A decline in fitness after midlife was also associated with dementia (OR, 3.0; 95% CI, 1.7-5.1), which was significant amongst both men and women and more pronounced in APOEε4 carriers (OR, 4.4; 95% CI, 2.1-9.1). Perceived poor physical fitness reflects a combination of biological and lifestyle-related factors that can increase dementia risk. A simple question about perceived physical fitness may reveal at-risk individuals who could benefit from preventive interventions.


Using Genetic Variation to Explore the Causal Effect of Maternal Pregnancy Adiposity on Future Offspring Adiposity: A Mendelian Randomisation Study.
It has been suggested that greater maternal adiposity during pregnancy affects lifelong risk of offspring fatness via intrauterine mechanisms. Our aim was to use Mendelian randomisation (MR) to investigate the causal effect of intrauterine exposure to greater maternal body mass index (BMI) on offspring BMI and fat mass from childhood to early adulthood. We used maternal genetic variants as instrumental variables (IVs) to test the causal effect of maternal BMI in pregnancy on offspring fatness (BMI and dual-energy X-ray absorptiometry [DXA] determined fat mass index [FMI]) in a MR approach. This was investigated, with repeat measurements, from ages 7 to 18 in the Avon Longitudinal Study of Parents and Children (ALSPAC; n = 2,521 to 3,720 for different ages). We then sought to replicate findings with results for BMI at age 6 in Generation R (n = 2,337 for replication sample; n = 6,057 for total pooled sample). In confounder-adjusted multivariable regression in ALSPAC, a 1 standard deviation (SD, equivalent of 3.7 kg/m2) increase in maternal BMI was associated with a 0.25 SD (95% CI 0.21-0.29) increase in offspring BMI at age 7, with similar results at later ages and when FMI was used as the outcome. A weighted genetic risk score was generated from 32 genetic variants robustly associated with BMI (minimum F-statistic = 45 in ALSPAC). The MR results using this genetic risk score as an IV in ALSPAC were close to the null at all ages (e.g., 0.04 SD (95% CI -0.21-0.30) at age 7 and 0.03 SD (95% CI -0.26-0.32) at age 18 per SD increase in maternal BMI), which was similar when a 97 variant generic risk score was used in ALSPAC. When findings from age 7 in ALSPAC were meta-analysed with those from age 6 in Generation R, the pooled confounder-adjusted multivariable regression association was 0.22 SD (95% CI 0.19-0.25) per SD increase in maternal BMI and the pooled MR effect (pooling the 97 variant score results from ALSPAC with the 32 variant score results from Generation R) was 0.05 SD (95%CI -0.11-0.21) per SD increase in maternal BMI (p-value for difference between the two results = 0.05). A number of sensitivity analyses exploring violation of the MR results supported our main findings. However, power was limited for some of the sensitivity tests and further studies with relevant data on maternal, offspring, and paternal genotype are required to obtain more precise (and unbiased) causal estimates. Our findings provide little evidence to support a strong causal intrauterine effect of incrementally greater maternal BMI resulting in greater offspring adiposity.


Normalization of serum rapid plasma reagin titer predicts normalization of cerebrospinal fluid and clinical abnormalities after treatment of neurosyphilis.
Success of neurosyphilis treatment is defined by normalization of cerebrospinal fluid (CSF) and clinical abnormalities. The goal of this study was to determine whether normalization of serum rapid plasma reagin (RPR) titer could accurately predict treatment success. One hundred ten patients who were enrolled in a longitudinal study of CSF abnormalities in syphilis had asymptomatic syphilitic meningitis, symptomatic syphilitic meningitis, or syphilitic eye disease and were treated for neurosyphilis. At 4, 7, and 13 months after treatment, serum RPR titer and CSF and clinical abnormalities were analyzed for normalization. Odds ratios for normalization of each CSF and clinical abnormality when serum RPR titer had normalized and the positive predictive value of normalization of serum RPR titer for normalization of CSF and clinical abnormalities were determined. Serum RPR titer had normalized in 63 patients (57%) by 4 months after treatment, in 94 (85%) by 7 months, and in 97 (88%) by 13 months. Except for CSF protein concentration, normalization of serum RPR titer predicted normalization of other CSF and clinical abnormalities in >80% of patients at 4 months, >85% at 7 months, and >90% at 13 months. The odds of normalization of CSF and clinical abnormalities were 28-57-fold higher when serum RPR titer had normalized, compared with when it had not. Normalization of serum RPR titer was consistently less accurate in predicting treatment success in human immunodeficiency virus-infected patients who were not receiving antiretroviral therapy, compared with those who were receiving such therapy. In most instances, normalization of serum RPR titer correctly predicts success of treatment of neurosyphilis, and follow-up lumbar puncture can be avoided.


Kcnj10 is a major type of K+ channel in mouse corneal epithelial cells and plays a role in initiating EGFR signaling.
We used primary mouse corneal epithelial cells (pMCE) to examine the role of Kcnj10 in determining membrane K(+) conductance and cell membrane potential and in regulating EGF/TGFA release. Western blot, immunostaining, and RT-PCR detected the expression of Kcnj10 in mouse cornea. The single channel recording identified the 20-pS inwardly rectifying K(+) channels in pMCE of WT mice, but these channels were absent in Kcnj10(-/-). Moreover, the whole cell recording demonstrates that deletion of Kcnj10 largely abolished the inward K(+) currents and depolarized the cell membrane K(+) reversal potential (an index of the cell membrane potential). This suggests that Kcnj10 is a main contributor to the cell K(+) conductance and it is pivotal in generating membrane potential in cornea. Furthermore, to test the hypothesis that Kcnj10 expression plays a key role in the stimulation of growth factors release, we employed an immortalized human corneal epithelial cell line (HCE) transfected with siRNA-Kcnj10 or siRNA-control. Levels of TGFA and EGF secreted in the medium were measured by ELISA. Coimmunoprecipitation, biotinylation, and pull-down assay were used to examine the expression of EGFR and the GTP bound form of Rac1 (active Rac1). Downregulation of Kcnj10 activated Rac1 and enhanced EGF/TGFA release, which further contributed to the upregulation of EGFR phosphorylation and surface expression. We conclude that Kcnj10 is a main K(+) channel expressed in corneal epithelial cells and the inhibition of Kcnj10 resulted in depolarization, which in turn induced an EGF-like effect.


Dysplasia and disorder of cell membrane entirety in iron-deficiency anemia.
Peripheral blood smears of 43 patients (26 males, median age 18 months, range: 6-180 months) with nutritional iron-deficiency anemia (IDA) were examined for the presence of trilineage hematological dysplasia. Twelve patients were reexamined for dysplastic findings after achieving a normal Hb and hematocrit level for age by the end of 2-3 months of iron treatment. A control group of 17 age-matched healthy children were also included. Neutrophils with loss of membrane entirety and protrusions were remarkable in 34/43 (79%) in the IDA group versus 1/12 (8%) after iron treatment and none of the control group. Microspherocytes were seen in 9/43 (21%) of IDA patients. Additionally, trilienage dysplasia was observed in the bone marrow samples available in 3 of the patients. It has been shown that iron-deficiency results in cellular DNA and RNA alterations, cell-cycle G1/S phase arrest, and apoptosis. Rac GTPases have been shown to control actin cytoskeleton, influencing cell polarity, microtubule dynamics, and the cytoskeletal organization of hematopoietic cells. Thus, the findings described above in neutrophils and red cells suggest a plausible link between iron and the Rac GTPase gene family. It may be a new avenue for iron waiting for proof.


Responses of Conversational Agents to Health and Lifestyle Prompts: Investigation of Appropriateness and Presentation Structures.
Conversational agents (CAs) are systems that mimic human conversations using text or spoken language. Their widely used examples include voice-activated systems such as Apple Siri, Google Assistant, Amazon Alexa, and Microsoft Cortana. The use of CAs in health care has been on the rise, but concerns about their potential safety risks often remain understudied. This study aimed to analyze how commonly available, general-purpose CAs on smartphones and smart speakers respond to health and lifestyle prompts (questions and open-ended statements) by examining their responses in terms of content and structure alike. We followed a piloted script to present health- and lifestyle-related prompts to 8 CAs. The CAs' responses were assessed for their appropriateness on the basis of the prompt type: responses to safety-critical prompts were deemed appropriate if they included a referral to a health professional or service, whereas responses to lifestyle prompts were deemed appropriate if they provided relevant information to address the problem prompted. The response structure was also examined according to information sources (Web search-based or precoded), response content style (informative and/or directive), confirmation of prompt recognition, and empathy. The 8 studied CAs provided in total 240 responses to 30 prompts. They collectively responded appropriately to 41% (46/112) of the safety-critical and 39% (37/96) of the lifestyle prompts. The ratio of appropriate responses deteriorated when safety-critical prompts were rephrased or when the agent used a voice-only interface. The appropriate responses included mostly directive content and empathy statements for the safety-critical prompts and a mix of informative and directive content for the lifestyle prompts. Our results suggest that the commonly available, general-purpose CAs on smartphones and smart speakers with unconstrained natural language interfaces are limited in their ability to advise on both the safety-critical health prompts and lifestyle prompts. Our study also identified some response structures the CAs employed to present their appropriate responses. Further investigation is needed to establish guidelines for designing suitable response structures for different prompt types.


Identification and characterization of DEDD2, a death effector domain-containing protein.
A novel Death Effector Domain-containing protein was identified, DEDD2, which is closest in amino acid sequence homology to death effector domain-containing DNA-binding protein, DEDD. DEDD2 mRNA is expressed widely in adult human tissues with highest levels in liver, kidney, and peripheral blood leukocytes. DEDD2 interacts with FLIP, but not with Fas-associated death domain (FADD) or caspase-8. Overexpression of DEDD2 induces moderate apoptosis and results in substantial sensitization to apoptosis induced by Fas (CD95/APO-1), tumor necrosis factor-related apoptosis-inducing ligand (TRAIL, Apo2L), or FADD. In contrast, Bax- or staurosporine-mediated cell death is not affected by expression of DEDD2. Fluorescence microscopy showed that overexpressed DEDD2 translocates to the nucleus, which is dependent on the presence of a bipartite nuclear localization signal in the DEDD2 protein. Mutagenesis studies revealed that the translocation of the DED of DEDD2 to the nucleus is essential for its pro-apoptotic activity. These findings suggest that DEDD2 is involved in the regulation of nuclear events mediated by the extrinsic apoptosis pathway.


Genome sequence of enterohaemorrhagic Escherichia coli O157:H7.
The bacterium Escherichia coli O157:H7 is a worldwide threat to public health and has been implicated in many outbreaks of haemorrhagic colitis, some of which included fatalities caused by haemolytic uraemic syndrome. Close to 75,000 cases of O157:H7 infection are now estimated to occur annually in the United States. The severity of disease, the lack of effective treatment and the potential for large-scale outbreaks from contaminated food supplies have propelled intensive research on the pathogenesis and detection of E. coli O157:H7 (ref. 4). Here we have sequenced the genome of E. coli O157:H7 to identify candidate genes responsible for pathogenesis, to develop better methods of strain detection and to advance our understanding of the evolution of E. coli, through comparison with the genome of the non-pathogenic laboratory strain E. coli K-12 (ref. 5). We find that lateral gene transfer is far more extensive than previously anticipated. In fact, 1,387 new genes encoded in strain-specific clusters of diverse sizes were found in O157:H7. These include candidate virulence factors, alternative metabolic capacities, several prophages and other new functions--all of which could be targets for surveillance.


Hypoxia and lineage specification of cell line-derived colorectal cancer stem cells.
Hypoxia is an important regulator of normal and cancer stem cell (CSC) differentiation. Colorectal CSCs from SW1222, LS180, and CCK81 colorectal cancer-derived cell lines are able to differentiate into complex 3D lumen-containing structures in normoxia, whereas in hypoxia, they form undifferentiated dense colonies that have reduced expression of the enterocyte differentiation marker CDX1, lack goblet cell formation, and have increased expression of BMI1 and activated Notch1. Hypoxia increases the clonogenicity of CSCs, which is cumulative as each round of hypoxia enriches for more CSCs. The hypoxic phenotype is reversible, because cells from hypoxic-dense colonies are able to reform differentiated structures when regrown in normoxia. We show that CDX1 is able to stimulate the generation of lumens even in hypoxia and has a negative feedback on BMI1 expression. Knockdown of CDX1 reduces lumen formation but does not affect goblet cell formation, suggesting that enterocytes and goblet cells form from different progenitor cells. Notch inhibition by dibenzazepine (DBZ) allowed CSCs to form goblet cells in both normoxia and hypoxia. Finally, we show that Hif1α, but not CA9, is an important mediator of the effects of hypoxia on the clonogenicity and differentiation of CSCs. In summary, hypoxia maintains the stem-like phenotype of colorectal cell line-derived CSCs and prevents differentiation of enterocytes and goblet cells by regulating CDX1 and Notch1, suggesting that this regulation is an important component of how hypoxia controls the switch between stemness and differentiation in CSCs.


Effects of elevated CO2 on predator avoidance behaviour by reef fishes is not altered by experimental test water.
Pioneering studies into the effects of elevated CO2 on the behaviour of reef fishes often tested high-CO2 reared fish using control water in the test arena. While subsequent studies using rearing treatment water (control or high CO2) in the test arena have confirmed the effects of high CO2 on a range of reef fish behaviours, a further investigation into the use of different test water in the experimental arena is warranted. Here, we used a fully factorial design to test the effect of rearing treatment water (control or high CO2) and experimental test water (control or high CO2) on antipredator responses of larval reef fishes. We tested antipredator behaviour in larval clownfish Amphiprion percula and ambon damselfish Pomacentrus amboinensis, two species that have been used in previous high CO2 experiments. Specifically, we tested if: (1) using control or high CO2 water in a two channel flume influenced the response of larval clownfish to predator odour; and (2) using control or high CO2 water in the test arena influenced the escape response of larval damselfish to a startle stimulus. Finally, (3) because the effects of high CO2 on fish behaviour appear to be caused by altered function of the GABA-A neurotransmitter we tested if antipredator behaviours were restored in clownfish treated with a GABA antagonist (gabazine) in high CO2 water. Larval clownfish reared from hatching in control water (496 µatm) strongly avoided predator cue whereas larval clownfish reared from hatching in high CO2 (1,022 µatm) were attracted to the predator cue, as has been reported in previous studies. There was no effect on fish responses of using either control or high CO2 water in the flume. Larval damselfish reared for four days in high CO2 (1,051 µatm) exhibited a slower response to a startle stimulus and slower escape speed compared with fish reared in control conditions (464 µatm). There was no effect of test water on escape responses. Treatment of high-CO2 reared clownfish with 4 mg l-1 gabazine in high CO2 seawater restored the normal response to predator odour, as has been previously reported with fish tested in control water. Our results show that using control water in the experimental trials did not influence the results of previous studies on antipredator behaviour of reef fishes and also supports the results of novel experiments conducted in natural reef habitat at ambient CO2 levels.


Characterization and photochemistry of 13-desmethyl bacteriorhodopsin.
The photochemistry of the 13-desmethyl (DM) analogue of bacteriorhodopsin (BR) is examined by using spectroscopy, molecular orbital theory, and chromophore extraction followed by conformational analysis. The removal of the 13-methyl group permits the direct photochemical formation of a thermally stable, photochemically reversible state, P1(DM) (lambda(max) = 525 nm), which can be generated efficiently by exciting the resting state, bR(DM) with yellow or red light (lambda > 590 nm). Chromophore extraction analysis reveals that the retinal configuration in P1(DM) is 9-cis, identical to that of the retinal configuration in the native BR P1 state. Fourier transform infrared and Raman experiments on P1(DM) indicate an anti configuration around the C15=N bond, as would be expected of an O-state photoproduct. However, low-temperature spectroscopy and ambient, time-resolved studies indicate that the P1(DM) state forms primarily via thermal relaxation from the L(D)(DM) state. Theoretical studies on the BR binding site show that 13-dm retinal is capable of isomerizing into a 9-cis configuration with minimal steric hindrance from surrounding residues, in contrast to the native chromophore in which surrounding residues significantly obstruct the corresponding motion. Analysis of the photokinetic experiments indicates that the Arrhenius activation energy of the bR(DM) --> P1(DM) transition in 13-dm-BR is less than 0.6 kcal/mol (vs 22 +/-5 kcal/mol measured for the bR --> P (P1 and P2) reaction in 85:15 glycerol:water suspensions of wild type). Consequently, the P1(DM) state in 13-dm-BR can form directly from all-trans, 15-anti intermediates (bR(DM) and O(DM)) or all-trans, 15-syn (K(D)(DM)/L(D)(DM)) intermediates. This study demonstrates that the 13-methyl group, and its interactions with nearby binding site residues, is primarily responsible for channeling one-photon photochemical and thermal reactions and is limited to the all-trans and 13-cis species interconversions in the native protein.


Studying in vitro membrane curvature recognition by proteins and its role in vesicular trafficking.
In recent years, the interest for proteins that exert key functions in vesicular trafficking through their ability to sense or induce positive membrane curvature has expanded. In this chapter, we first present simple protocols to determine whether a protein targets positively curved membranes with liposomes of well-defined size. Next we describe more sophisticated approaches based on the controlled deformation of giant liposomes. These approaches allow visualization and quantification of protein binding to membrane regions of high curvature by real-time fluorescence microscopy. Last we describe several functional assays to measure how membrane curvature controls the activation state of Arf1 via ArfGAP1 or the asymmetric tethering between flat and curved membranes via the golgin GMAP-210.


First Report of Fusarium Rot Caused by Fusarium oxysporum on Lemon in Tucumán, Argentina.
Fusarium rot is considered a minor disease of citrus fruits. Several Fusarium species have been associated with fruit decay, most commonly F. lateritium Nees, F. moniliforme J. Sheld., F. oxysporum Schltdl., and F. solani (Mart.) Sacc. (2,3). In the winters of 2007, 2009, 2010, and 2011, lemon [Citrus limon (L.) Burm. f.] fruit with white mycelium covering the peduncle were submitted to the Phytopathology Lab at the Estación Experimental Agroindustrial Obispo Colombres. All fruit samples from Tucumán, Argentina, were stored in boxes kept in packinghouse for more than 1 month. In 2007 only, light to dark brown flavedo around the peduncle was observed in less than 1% of the sample fruit received. No internal breakdown was visible. No change in rind color was observed in the samples received in remaining years. Abundant Fusarium sp. conidia were observed on the mycelium. Colonies with white to violet fluffy aerial mycelium developed on potato dextrose agar (PDA) and produced abundant ovoid or oblong microconidia (1.9 to 3.6 × 4.8 to 10.8 μm), usually unicellular, borne in false heads on short monophialides, and loculated slightly falcate macroconidia were mostly three to five septate (2.4 to 4.8 × 19.2 to 31.2 μm). Unbranched and branched-monophialidic conidiophores were observed. Simple or paired chlamydospores developed on synthetic nutrient agar (1 g KH2PO4, 1 g KNO3, 0.5 g MgSO4.7H2O, 0.5 g KCl, 0.2 g sucrose, and 20 g agar/liter distilled water). On the basis of morphological and cultural criteria, 22 isolates were identified as F. oxysporum (4) designated as D1 to D22. Morfological identification was confirmed by PCR (1) using genomic DNA extracted from the mycelium of pure culture, and an amplified product of 70 bp, specific for the species F. oxysporum, was obtained. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS5 and secuenced. BLAST analysis of the 600 bp segment showed a 100% indentity with F. oxysporum, strains CCF 4362 and 1166 (GenBank Accession Nos. HE974454 and FR731133, respectively). Pathogenicity tests were conducted twice by inoculating 10 surface-disinfected wounded lemon fruit. A rind disc (5 mm in diameter and 1 mm deep) near the stem end was removed and a 5-mm-diameter agar disc of D2 isolate (grown at 25°C for 5 days on PDA) was attached to the wound replacing the rind disc. The inoculation site was covered with moistened cotton wool and the fruit were wrapped in plastic bags to prevent the inoculum from drying out. Ten control fruit were inoculated with uncultured PDA plugs (5 mm in diameter). All fruit were maintained in a growth chamber at 25°C under humid conditions. After 5 to 6 days, all inoculated fruit showed white aerial mycelium, initially on the inoculation site and then on the peduncle, similar to that observed on naturally infected fruit. After 20 days, two fruit developed stem end dry rot and showed peduncle fall but no internal breakdown was visible. Control fruit developed any symptom as described above. F. oxysporum was consistently reisolated from infected tissues, completing Koch's postulates. To our knowledge, this is the first report of Fusarium rot caused by F. oxysporum on lemon in Tucumán, Argentina. References: (1) V. Edel et al. Mycol. Res. 104:518, 2000. (2) H. S. Fawcett. Citrus Diseases and Their Control, 1936. (3) A. Z. Joffe and M. Schiffmann-Nadel. Fruits 27:117, 1972. (4) P. E. Nelson et al. Fusarium species: An Illustrated Manual for Identification, 1983.


Attention to locations and features: different top-down modulation of detector weights.
It is well known that attention improves the visibility of a target. In this study, we examined the effect of attention on the selectivity profile for a target. We used a masking technique to measure the tuning function for detecting a target while cueing either its orientation or its location. In the presence of an orientation mask, uncued thresholds were maximally elevated with a parallel mask and decreased with increasing mask orientation from the target. The presence of a cue reduced the masking effect but the shape of the function was cue-specific: The orientation cue consistently improved thresholds at the target orientation, whereas the location cue typically improved thresholds at all orientations relative to the function measured in the absence of attention. The selective versus overall increase of sensitivity observed in our study may be due to differences in the weighting of individual detectors that determine the behavioral tuning function in the two cueing conditions.


Measurement of long focal lengths with a double-grating interferometer.
A method for measuring long focal lengths using a double-grating interferometer is proposed. The intensity distribution of a double-grating interferometer illuminated by a spherical beam is derived with diffraction theory. A tiny rotation angle is set between the two gratings, and a high-contrast interference pattern is produced by the adjacent diffracted orders. The angular change in the fringes after insertion of a test lens is a measure of the focal length. The uncertainty due to aberration of the collimated beam was analyzed by measuring a series of lenses. The relative deviations are less than 0.1%.


Comparing expression profiles of genes with similar promoter regions.
Gene regulatory elements are often predicted by seeking common sequences in the promoter regions of genes that are clustered together based on their expression profiles. We consider the problem in the opposite direction: we seek to find the genes that have similar promoter regions and determine the extent to which these genes have similar expression profiles. We use the data sets from experiments on Saccharomyces cerevisiae. Our similarity measure for the promoter regions is based on the set of common mapped or putative transcription factor binding sites and other regulatory elements in the upstream region of the genes, as contained in the Saccharomyces cerevisiae Promoter Database. We pair up the genes with high similarity scores and compare their expression levels in time-course experiment data. We find that genes with similar promoter regions on the average have significantly higher correlation, but it can vary widely depending on the genes. This confirms that the presence of similar regulatory elements often does not correspond to similarity in expression profiles and indicates that finding transcription factor binding sites or other regulatory elements starting with the expression patterns may be limited in many cases. Regardless of the correlation, the degree to which the profiles agree under different experimental conditions can be examined to derive hypotheses concerning the role of common regulatory elements. Overall, we find that considering the relationship between the promoter regions and the expression profiles starting with the regulatory elements is a difficult but useful process that can provide valuable insights.


Research Priorities on the Relationship between Wasting and Stunting.
Wasting and stunting are global public health problems that frequently co-exist. However, they are usually separated in terms of policy, guidance, programming and financing. Though both wasting and stunting are manifestations of undernutrition caused by disease and poor diet, there are critical gaps in our understanding of the physiological relationship between them, and how interventions for one may affect the other. The aim of this exercise was to establish research priorities in the relationships between wasting and stunting to guide future research investments. We used the CHNRI (Child Health and Nutrition Research Initiative) methodology for setting research priorities in health. We utilised a group of experts in nutrition, growth and child health to prioritise 30 research questions against three criteria (answerability, usefulness and impact) using an online survey. Eighteen of 25 (72%) experts took part and prioritised research directly related to programming, particularly at the public health level. The highest-rated questions were: "Can interventions outside of the 1000 days, e.g. pre-school, school age and adolescence, lead to catch-up in height and in other developmental markers?"; "What timely interventions work to mitigate seasonal peaks in both wasting and stunting?"; and "What is the optimal formulation of ready-to-use foods to promote optimal ponderal growth and also support linear growth during and after recovery from severe acute malnutrition?" There was a high level of agreement between experts, particularly for the highest ranking questions. Increased commitment to rigorous evaluations of treatment and prevention interventions at the public health level, addressing questions of the timing of intervention, and the extent to which impacts for both wasting and stunting can be achieved, is needed to inform global efforts to tackle undernutrition and its consequences.


Treatment outcomes for HIV and MDR-TB co-infected adults and children: systematic review and meta-analysis.
The incidence of multidrug-resistant tuberculosis (MDR-TB) is increasing in high human immunodeficiency virus (HIV) prevalence settings, with high associated mortality. Treatment outcomes in HIV-co-infected adults and children are poorly documented. To systematically assess treatment outcomes among HIV-MDR-TB co-infected patients. We searched two databases and the proceedings of an annual international conference up to November 2014 for studies reporting on major clinical outcomes among HIV-MDR-TB-co-infected adults and children, and pooled the results using random-effects meta-analysis. Of 4812 abstracts and articles screened, 30 studies providing data on 2578 adults and 147 children were included. Overall pooled treatment success was 56.9% (95% confidence interval [CI] 46.2-67.6), 49.9% (95%CI 38.5-61.2) among adults and 83.4% (95%CI 74.7-92) among children. Mortality was 38% in adults (95%CI 28-48.1) and 11.4% (95%CI 5.8-17.1) in children. Loss to follow-up was higher among adults (16.1%, 95%CI 9-23.2) than among children (3.9%, 95%CI 0.9-6.9). Adverse events were experienced by the majority of patients; however, this was inconsistently documented. The use of fluoroquinolones, aminoglycosides and Group IV drugs appeared to be associated with treatment success. The proportion of HIV-MDR-TB-co-infected patients achieving treatment success was similar to success rates reported among MDR-TB patients in general, regardless of HIV status; however, mortality was higher, particularly among adults, highlighting the need for early diagnosis and more effective treatment regimens.


Phenotypes Associated with Pathogenicity: Their Expression in Arctic Fungal Isolates.
Around 85% of the environments on Earth are permanently or seasonally colder than 5 °C. Among those, the poles constitute unique biomes, which harbor a broad variety of microbial life, including an abundance of fungi. Many fungi have an outstanding ability to withstand extreme conditions and play vital ecosystem roles of decomposers as well as obligate or facultative symbionts of many other organisms. Due to their dispersal capabilities, microorganisms from cryosphere samples can be distributed around the world. Such dispersal involves both species with undefined pathogenicity and potentially pathogenic strains. Here we describe the isolation of fungal species from pristine Arctic locations in Greenland and Svalbard and the testing of the expression of characteristics usually associated with pathogenic species, such as growth at 37 °C, hemolytic ability, and susceptibility to antifungal agents. A total of 320 fungal isolates were obtained, and 24 of the most abundant and representative species were further analyzed. Species known as emerging pathogens, like Aureobasidium melanogenum, Naganishia albida, and Rhodotorula mucilaginosa, were able to grow at 37 °C, showed beta-hemolytic activity, and were intrinsically resistant to commonly used antifungals such as azoles and echinocandins. Antifungal resistance screening revealed a low susceptibility to voriconazole in N. albida and Penicillium spp. and to fluconazole in Glaciozyma watsonii and Glaciozyma-related taxon.


The evolutionary fate of the horizontally transferred agrobacterial mikimopine synthase gene in the genera Nicotiana and Linaria.
Few cases of spontaneously horizontally transferred bacterial genes into plant genomes have been described to date. The occurrence of horizontally transferred genes from the T-DNA of Agrobacterium rhizogenes into the plant genome has been reported in the genus Nicotiana and in the species Linaria vulgaris. Here we compare patterns of evolution in one of these genes (a gene encoding mikimopine synthase, mis) following three different events of horizontal gene transfer (HGT). As this gene plays an important role in Agrobacterium, and there are known cases showing that genes from pathogens can acquire plant protection function, we hypothesised that in at least some of the studied species we will find signs of selective pressures influencing mis sequence. The mikimopine synthase (mis) gene evolved in a different manner in the branch leading to Nicotiana tabacum and N. tomentosiformis, in the branch leading to N. glauca and in the genus Linaria. Our analyses of the genus Linaria suggest that the mis gene began to degenerate soon after the HGT. In contrast, in the case of N. glauca, the mis gene evolved under significant selective pressures. This suggests a possible role of mikimopine synthase in current N. glauca and its ancestor(s). In N. tabacum and N. tomentosiformis, the mis gene has a common frameshift mutation that disrupted its open reading frame. Interestingly, our results suggest that in spite of the frameshift, the mis gene could evolve under selective pressures. This sequence may still have some regulatory role at the RNA level as suggested by coverage of this sequence by small RNAs in N. tabacum.


Examining reward-seeking, negative self-beliefs and over-general autobiographical memory as mechanisms of change in classroom prevention programs for adolescent depression.
Effective methods to prevent adolescent depressive symptoms could reduce suffering and burden across the lifespan. However, psychological interventions delivered to adolescents show efficacy only in symptomatic or high-risk youth. Targeting causal risk factors and assessing mechanistic change can help devise efficacious universal or classroom based prevention programs. A non-randomized longitudinal design was used to compare three classroom-based prevention programs for adolescent depression (Behavioral Activation with Reward Processing, "Thinking about Reward in Young People" (TRY); Cognitive Behavioral Therapy (CBT) and Mindfulness Based Cognitive Therapy (MBCT)), and determine cognitive mechanisms of change in these programs. Cognitive mechanisms examined were reward-seeking, negative self-beliefs (assessed with behavioral tasks) and over-general autobiographical memory. 256 healthy adolescents aged 13-14 participated with 236 (92%) and 227 (89%) completing the pre- and post-assessments. TRY was the only intervention associated with a reduction in depressive symptoms at follow-up. Reward-seeking increased following TRY. In the other programs there were non-significant changes in cognitive mechanisms, with more reflective negative self-beliefs in CBT and fewer over-general autobiographical memories in MBCT In the TRY program, which focused on increasing sensitivity to rewarding activities, reward seeking increased and this was associated with decreased depressive symptoms. Due to the infeasibility of a cluster randomized controlled trial, a non-randomized design was used. Increased reward-seeking was associated with decreased depressive symptoms and may be a mechanism of depressive symptom change in the intervention with a focus on enhancing sensitivity and awareness of reward. This study provides preliminary evidence to suggest that incorporating activities to enhance reward sensitivity may be fruitful in randomized controlled trials of universal prevention programs for depression.


Perspectives on Principles of Cellular Behavior from The Biophysics of Protists.
Cells are the fundamental unit of biological organization. Although it may be easy to think of them as little more than the simple building blocks of complex organisms such as animals, single cells are capable of behaviors of remarkable apparent sophistication. This is abundantly clear when considering the diversity of form and function among the microbial eukaryotes, the protists. How might we navigate this diversity in the search for general principles of cellular behavior? Here, we review cases in which the intensive study of protists from the perspective of cellular biophysics has driven insight into broad biological questions of morphogenesis, navigation and motility, and decision making. We argue that applying such approaches to questions of evolutionary cell biology presents rich, emerging opportunities. Integrating and expanding biophysical studies across protist diversity, exploiting unique characteristics of each organism, will enrich our understanding of general, underlying principles.


Osteopathic manipulative treatment in pudendal neuralgia: A case report.
Pudendal neuralgia is characterised by pain in the pudendal dermatome. It could be due to a stenosis of the pudendal canal, a compression along its pathway, or a pelvic trauma. Pudendal nerve entrapment (PNE) syndrome is frequently involved in pudendal neuralgia onset. This case report describes the osteopathic manipulative treatment (OMT) of a patient with functional PNE. A 40-year-old female presented with a 12-month history of intense pelvic pain resistant to 3 months of pharmacologic treatment that arose after three proctological surgeries. A perineal retracted painful scar was visible upon examination. PNE syndrome diagnosis was based on Nantes criteria. The electromyogram of the nerve showed an increased motor response latency of the left pudendal nerve. Visual analogue scale (VAS), female National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI), Oswestry Disability Index (ODI) and Tampa scale of kinesiophobia (TSK) were used to assess patient's symptoms at baseline (T0), after pharmacologic treatment (T1), after OMT (T2), and at 6-month follow-up. Five treatments, including direct and indirect techniques, were performed over 1 month. OMT reduced pelvic neuralgia and disability indexes without any complications, maintaining a positive outcome at 6-month follow-up (VAS: T0 = 10, T1 = 10, T2 = 1.8, T3 = 1.5), (NIH-CPSI: T0 = 34, T1 = 30, T2 = 7, T3 = 6), (ODI: T0 = 48, T1 = 29, T2 = 9, T3 = 5) and (TSK: T0 = 51, T1 = 41, T2 = 20, T3 = 17). This is the first report of a patient diagnosed with functional PNE managed with OMT. A link between PNE, scar and pelvic somatic dysfunctions could suggest double crush syndrome.


SLC2A9 (GLUT9) mediates urate reabsorption in the mouse kidney.
Uric acid (UA) is a metabolite of purine degradation and is involved in gout flairs and kidney stones formation. GLUT9 (SLC2A9) was previously shown to be a urate transporter in vitro. In vivo, humans carrying GLUT9 loss-of-function mutations have familial renal hypouricemia type 2, a condition characterized by hypouricemia, UA renal wasting associated with kidney stones, and an increased propensity to acute renal failure during strenuous exercise. Mice carrying a deletion of GLUT9 in the whole body are hyperuricemic and display a severe nephropathy due to intratubular uric acid precipitation. However, the precise role of GLUT9 in the kidney remains poorly characterized. We developed a mouse model in which GLUT9 was deleted specifically along the whole nephron in a tetracycline-inducible manner (subsequently called kidney-inducible KO or kiKO). The urate/creatinine ratio was increased as early as 4 days after induction of the KO and no GLUT9 protein was visible on kidney extracts. kiKO mice are morphologically identical to their wild-type littermates and had no spontaneous kidney stones. Twenty-four-hour urine collection revealed a major increase of urate urinary excretion rate and of the fractional excretion of urate, with no difference in urate concentration in the plasma. Polyuria was observed, but kiKO mice were still able to concentrate urine after water restriction. KiKO mice displayed lower blood pressure accompanied by an increased heart rate. Overall, these results indicate that GLUT9 is a crucial player in renal handling of urate in vivo and a putative target for uricosuric drugs.


Multidrug resistance in breast cancer: a meta-analysis of MDR1/gp170 expression and its possible functional significance.
P-glycoprotein (gp170; encoded by the MDR1 gene [also known as PGY1]) is a membrane protein capable of exporting a variety of anticancer drugs from cells. MDR1/gp170 expression has been studied in breast cancer, but the prevalence of this expression and its role in breast tumor drug resistance are unclear. We conducted a critical review and meta-analysis of studies examining MDR1/gp170 expression in breast cancer to estimate the likely prevalence and clinical relevance of this expression. We also explored reasons for differences in the findings from individual studies. Published papers on MDR1/gp170 expression in breast cancer were identified by searching several literature databases and reviewing the bibliographies of identified papers. Variability across the studies in the proportion of tumors expressing MDR1/gp170 was assessed by use of chi-squared tests of homogeneity, weighted means, and weighted linear regression. Pooled relative risks (RRs) for the association between the induction of MDR1/gp170 expression and prior chemotherapy and associations between MDR1/gp170 expression and several clinical outcomes were estimated by use of Mantel-Haenszel methods. Heterogeneity among the pooled RRs was explored by use of chi-squared tests. Reported P values are two-sided. Thirty-one studies were identified and evaluated. The proportion of breast tumors expressing MDR1/gp170 in all of the studies was 41.2%, but there was substantial heterogeneity in the values across individual studies (P<.0001). Regression analyses demonstrated that a considerable portion of the observed heterogeneity was a consequence of the change, over time, from RNA hybridization-based assays to immunohistochemistry-based assays of MDR1/gp170 expression. Measuring MDR1/gp170 expression before versus after chemotherapy and use of cytotoxic drugs that are not substrates for gp170 also contributed to the heterogeneity. Treatment with chemotherapeutic drugs or hormonal agents was associated with an increase in the proportion of tumors expressing MDR1/gp170 (RR = 1.77; 95% confidence interval [CI] = 1.46-2.15). Patients with tumors expressing MDR1/gp170 were three times more likely to fail to respond to chemotherapy than patients whose tumors were MDR1/gp170 negative (RR = 3.21; 95% CI = 2.28-4.51); this RR increased to 4.19 (95% CI = 2.71-6.47) when considering only patients whose tumor expression of MDR1/gp170 was measured after chemotherapy. MDR1/gp170 expression was not associated with lymph node metastases, estrogen receptor status, tumor size, tumor grade, or tumor histology. MDR1/gp170 expression in breast tumors is associated with treatment and with a poor response to chemotherapy. The data are consistent with a contributory role for MDR1/gp170 in the multidrug resistance in some breast tumors.


Decentering as a core component in the psychological treatment and prevention of youth anxiety and depression: a narrative review and insight report.
Decentering is a ubiquitous therapeutic concept featuring in multiple schools of psychological intervention and science. It describes an ability to notice to day-to-day psychological stressors (negative thoughts, feelings, and memories) from an objective self-perspective and without perseverating on the themes they represent. Thus, decentering dampens the impact and distress associated with psychological stressors that can otherwise increase mental ill health in vulnerable individuals. Importantly, the strengthening of decentering-related abilities has been flagged as a core component of psychological interventions that treat and prevent anxiety and depression. We provide an in-depth review evidence of the salutary effects of decentering with a special focus on youth mental health. This is because adolescence is a critical window for the development of psychopathology but is often under-represented in this research line. A narrative synthesis is presented that integrates and summarizes findings on a range of decentering-related abilities. Section 1 reviews extant conceptualizations of decentering and data-driven approaches to characterize its characteristic. A novel definition is then offered to guide future empirical research. Section 2 overviews laboratory-based research into the development of decentering as well as its relationship with anxiety and depression. Section 3 examines the role decentering-related skills play in psychological interventions for anxiety and depression. Critically, we review evidence that treatment-related increases in decentering predict latter reductions in anxiety and depression severity. Each section highlights important areas for future research. The report concludes by addressing the vital questions of whether, how, why and when decentering alleviates youth anxiety and depression.


Views of UK-trained medical graduates of 1999-2009 about their first postgraduate year of training: national surveys.
In the UK, doctors' first year of medical work is also their first year of postgraduate training. It is very important that their experience of work and training is good. Surveys of entire cohorts graduating in particular years. UK. Questionnaires sent 1 year after qualification to all UK medical graduates of 1999, 2000, 2002, 2005, 2008 and 2009. The study comprised 17 831 respondents. Variation in views across cohorts was modest. Overall, 30% agreed their training had been of a high standard; 38% agreed educational opportunities had been good; 52% agreed they had to do too much routine non-medical work; and 16% agreed they had to perform clinical tasks for which they felt inadequately trained. Job enjoyment, rated from 1 ('I didn't enjoy it at all') to 10 ('I enjoyed it greatly'), improved from 70% of doctors in the 1999 cohort scoring 7-10 to 75% in the 2009 cohort. Satisfaction with available leisure time, rated from 1 ('not at all satisfied') to 10 ('extremely satisfied'), rose from 24% scoring 7-10 in the 1999s to 49% in the 2009s. Male-female differences were small. There was improvement over the decade in some aspects of work, particularly satisfaction with time off work for leisure, and overall enjoyment of the job. There was little change in doctors' views about the training experience offered by the F1 year.


Exploration of Circadian Rhythms in Patients with Bilateral Vestibular Loss.
New insights have expanded the influence of the vestibular system to the regulation of circadian rhythmicity. Indeed, hypergravity or bilateral vestibular loss (BVL) in rodents causes a disruption in their daily rhythmicity for several days. The vestibular system thus influences hypothalamic regulation of circadian rhythms on Earth, which raises the question of whether daily rhythms might be altered due to vestibular pathology in humans. The aim of this study was to evaluate human circadian rhythmicity in people presenting a total bilateral vestibular loss (BVL) in comparison with control participants. Nine patients presenting a total idiopathic BVL and 8 healthy participants were compared. Their rest-activity cycle was recorded by actigraphy at home over 2 weeks. The daily rhythm of temperature was continuously recorded using a telemetric device and salivary cortisol was recorded every 3 hours from 6:00AM to 9:00PM over 24 hours. BVL patients displayed a similar rest activity cycle during the day to control participants but had higher nocturnal actigraphy, mainly during weekdays. Sleep efficiency was reduced in patients compared to control participants. Patients had a marked temperature rhythm but with a significant phase advance (73 min) and a higher variability of the acrophase (from 2:24 PM to 9:25 PM) with no correlation to rest-activity cycle, contrary to healthy participants. Salivary cortisol levels were higher in patients compared to healthy people at any time of day. We observed a marked circadian rhythmicity of temperature in patients with BVL, probably due to the influence of the light dark cycle. However, the lack of synchronization between the temperature and rest-activity cycle supports the hypothesis that the vestibular inputs are salient input to the circadian clock that enhance the stabilization and precision of both external and internal entrainment.


Measuring detachment of Aspergillus niger spores from colonies with an atomic force microscope.
Detachment of fungal spores from moldy surfaces and the subsequent aerosolization can lead to adverse health effects. Spore aerosolization occurs when the forces for aerosolization exceed the binding forces of spores with their colonies. The threshold force to detach a spore from a growing colony remains unknown. This investigation measured the detachment of spores of Aspergillus niger from a colony using an atomic force microscope (AFM). The spores were first affixed to the cantilever of the AFM with ultraviolet curing glue, and then, the colony was moved downward until the spores detached. The threshold detachment forces were inferred from the deflection of the cantilever. In addition, the spores were aerosolized in a wind tunnel by a gradual increase of the blowing air speed. The forces measured by the AFM were compared with the hydrodynamic forces for aerosolization. The AFM measurements revealed that a force of 3.27 ± 0.25 nN was required to detach a single spore from the 4-day-old colony, while 1.98 ± 0.13 nN was sufficient for the 10-day-old colony. Slightly smaller detachment forces were observed by the AFM than were determined by the aerosolization tests.


Mild and repetitive very mild axonal stretch injury triggers cystoskeletal mislocalization and growth cone collapse.
Diffuse axonal injury is a hallmark pathological consequence of non-penetrative traumatic brain injury (TBI) and yet the axonal responses to stretch injury are not fully understood at the cellular level. Here, we investigated the effects of mild (5%), very mild (0.5%) and repetitive very mild (2×0.5%) axonal stretch injury on primary cortical neurons using a recently developed compartmentalized in vitro model. We found that very mild and mild levels of stretch injury resulted in the formation of smaller growth cones at the tips of axons and a significantly higher number of collapsed structures compared to those present in uninjured cultures, when measured at both 24 h and 72 h post injury. Immunocytochemistry studies revealed that at 72 h following mild injury the axonal growth cones had a significantly higher colocalization of βIII tubulin and F-actin and higher percentage of collapsed morphology than those present following a very mild injury. Interestingly, cultures that received a second very mild stretch injury, 24 h after the first insult, had a further increased proportion of growth cone collapse and increased βIII tubulin and F-actin colocalization, compared with a single very mild injury at 72 h PI. In addition, our results demonstrated that microtubule stabilization of axons using brain penetrant Epothilone D (EpoD) (100 nM) resulted in a significant reduction in the number of fragmented axons following mild injury. Collectively, these results suggest that mild and very mild stretch injury to a very localized region of the cortical axon is able to trigger a degenerative response characterized by growth cone collapse and significant abnormal cytoskeletal rearrangement. Furthermore, repetitive very mild stretch injury significantly exacerbated this response. Results suggest that axonal degeneration following stretch injury involves destabilization of the microtubule cytoskeleton and hence treatment with EpoD reduced fragmentation. Together, these results contribute a better understanding of the pathogenesis of mild and repetitive TBI and highlight the therapeutic effect of microtubule targeted drugs on distal part of neurons using a compartmentalized culturing model.


Decline of trace metal pollution in the bottom sediments of the Barcelona City continental shelf (NW Mediterranean).
The evolution of trace metal pollution on the Barcelona city continental shelf during the last few decades was studied by analyzing the historical records of trace metals in sediment cores and surface sediment samples taken at the same locations in 1987 and in 2008. Polluted surface samples taken in 1987 reached enrichment factors of up to 490 for Hg, about 40 for Pb and Cd, and about 17 for Zn, Cr and Cu. The data show a decline of up to one order of magnitude in the trace metal content of surface sediments during the last few decades, with maximum enrichment factors of between 20 and 30 for Hg and Cd and between 5 and 12 for Zn, Cr, Pb and Cu. Although present-day pollution is still significant, it is evident that environmental regulations that are in place, including the operation since 1979 of wastewater treatment plants built in the Besòs River watershed, have drastically reduced the pollution levels in this highly populated and industrialized Mediterranean area. However, water discharge during heavy rain events exceeds the treatment capacity of the existing facilities, leading to the maintenance of still high levels of metals in sediments of the Barcelona city shelf.


Evaluation of body mass index and related lifestyle factors among 14-17-year-old Turkish adolescents.
During adolescence, unhealthy body weight status is considered as a global concern as it may lead to adverse health consequences in adulthood, therefore evaluation of the risk factors is crucial. The aim of the study was to determine the prevalence of adolescents under the risk of being underweight, overweight, and obese among 14-17-year-old Turkish adolescents. In addition, we examined the association between unhealthy body weight categories and lifestyle factors. This study was designed as cross-sectional study which included body weight status and associated parameters of 1561 adolescents aged between 14 and 17 who were registered 25 different high schools in Istanbul, Uskudar. Height and body weight of participants were measured and related factors were obtained through a questionnaire. Differences in distributions were analyzed using the Chi-square test and to control confounding factors, multivariate logistic regression analysis was performed. As statistical significance limit of p<0.05 was determined. Body mass index (BMI) percentile analyses indicated that 3.6% of participants were underweight, 14.3% were overweight, and 13.8% were obese. We demonstrated that age, gender, and school types were statistically very significantly associated with BMI (p<0.001) and daily meal frequency, eating speed, and mealtime regularity were significantly related with BMI (p<0.05). In addition, according to multivariate analysis results, gender and school types were closely related with obesity among 14-17-year old Turkish adolescents. This study has been demonstrated that frequency of adolescent obesity in Istanbul, Turkey, has increased and obesity closely related to gender and type of school. Further educational and interventional studies should be organized in this region with consideration of relevant risk factors.


Testosterone and cortisol levels in university students reflect actual rather than estimated number of wrong answers on written exam.
After dominance-related encounters, testosterone levels increase in winners and decrease in losers. In humans, many exceptions have been described. It is possible that the complicated patterns in humans result from the methods limitations--measurement of hormone concentrations in simulated competitive events or sport instead in real-life situations. Here we studied changes in hormonal levels and self-estimated attractivity in real situations, namely in students after written exams. We observed that the testosterone and cortisol increased or decreased in relation to the number of wrong answers on the exam. The number of wrong answers was a better predictor of the hormonal changes (increase of both testosterone and cortisol in successful, decrease in unsuccessful students) than the self-estimated number of wrong answers or a subjectively opinionated impression from the exam. On the contrary, the concentration of hormones before the exam and self-estimated attractivity were better predictors of the subjective impression from the exam than the number of wrong answers. Our results suggest that the students' subconsciousness, which directly influences the concentration of hormones, is able to objectively estimate results of an exam better than their consciousness.


Inflorescence spiders: A cost/benefit analysis for the host plant, Haplopappus venetus Blake (Asteraceae).
Predators on flower visitors, such as spiders, could influence plant reproduction by determining the balance between pollination and seed predation by insects. This study examines the net effect of predation by the inflorescence spider, Peucetia viridans (Hentz), for seed production by a native plant species on which it hunts. Both pollination and seed set of Haplopappus venetus (Asteraceae) were reduced on branches with spiders; however, the release of viable, undamaged seed was higher on inflorescence branches with spiders than on those without. Occurrence of P. viridans was associated with the flat-topped inflorescence branch structure characteristic of H. venetus rather than with the vertical structure of its congener, H. squarrosus. Thus, the interaction should be a reinforcing selective pressure on inflorescence branch morphology of H. venetus over time. Two factors providing constraints on the degree and rate of coevolution of the plant-spider interaction are suggested by the results: (1) the critical role of phenological synchrony and (2) the opposing requirements of interacting species and of subsequent life history stages within a species.


Prolonged alteration in E-box binding after a single systemic kainate injection: potential relation to F1/GAP-43 gene expression.
The presence in hippocampus of a basic helix-loop-helix (bHLH) family of transcription factors (TFs) specifically binding in an electrophoretic mobility shift assay (EMSA) to the E-box recognition element was established by selective blockade of binding both by cold competition and by an antibody to MyoD1, an E-box TF. Protein source was from a micro-dissected preparation enriched in hippocampal granule cells. Specific E-box binding of hippocampal transcription factors was significantly reduced in kainate acid (KA) treated animals. This was observed at 24 and 72 h, but not before (3, 6 h) or after (96 h). This is the first report to our knowledge to study functional regulation of E-box binding protein in adult hippocampus. To determine the generality of this E-box regulatory event, we studied four other situations, in addition to kainate treatment, where axonal growth is known or has been suggested to increase: NGF treatment of PC12 cells, unilateral hilar lesions, long-term potentiation after 1 h, and postnatal rat hippocampal development. In all four cases, decreased E-box binding was observed. The recent link of F1/GAP-43 mRNA induction in hippocampal granule cells by KA to growth of their axons, the mossy fibers in the adult rat, suggests a potential role for the F1/GAP-43 5' flanking promoter region in regulating neurite outgrowth. Since in all cases decreased E-box binding preceded increased F1/GAP-43 mRNA expression, it is suggested that E-box binding to the F1/GAP-43 promoter in hippocampal granule cells could negatively regulate F1/GAP-43 gene expression. Indeed, analysis of recognition elements on the F1/GAP-43 gene revealed an arrangement, previously described in other genes, of multiple adjacent E-box elements. E-box binding of bHLH transcription factors is likely to occur on several different genes in addition to F1/GAP-43. It is, therefore, attractive to think that E-box binding is regulated by in vivo activation of the adult brain and that this gene regulatory event participates in the orchestration of molecular and cellular responses underlying axonal growth.


Cannabidiol and Cannabigerol Inhibit Cholangiocarcinoma Growth In Vitro via Divergent Cell Death Pathways.
Cholangiocarcinoma (CCA) is a rare and highly lethal disease with few effective treatment options. Cannabinoids, cannabidiol (CBD) and cannabigerol (CBG) are non-psychedelic components extracted from cannabis. These non-psychoactive compounds have shown anti-proliferative potential in other tumor models; however, the efficacy of CBD and CBG in CCA is unknown. Furthermore, two cell death pathways are implicated with CBD resulting in autophagic degeneration and CBG in apoptosis. HuCC-T1 cells, Mz-ChA-1 cells (CCA cell lines) and H69 cells (immortalized cholangiocytes), were treated with CBD and CBG for 24 to 48 h. The influence of these cannabinoids on proliferation was assessed via MTT assay. Apoptosis and cell cycle were evaluated via Annexin-V apoptosis assay and propidium iodide, respectively. The expression of proliferation biomarker Ki-67, apoptosis biomarker BAX, and autophagic flux biomarkers LC3b and LAMP1 were evaluated via immunofluorescence. Cell migration and invasion were evaluated via wound healing assay and trans-well migration invasion assays, respectively. The colony formation was evaluated via colony formation assay. In addition, the expression of autophagy gene LC3b and apoptosis genes BAX, Bcl-2, and cleaved caspase-3 were evaluated via Western blot. CBD and CBG are non-selective anti-proliferative agents yielding similar growth curves in CCA; both cannabinoids are effective, yet CBG is more active at lower doses. Low doses of CBD and CBG enhanced immortalized cholangiocyte activity. The reduction in proliferation begins immediately and occurs maximally within 24 h of treatment. Moreover, a significant increase in the late-stage apoptosis and a reduction in the number of cells in S stage of the cell cycle indicates both CBD and CBG treatment could promote apoptosis and inhibit mitosis in CCA cells. The fluorescent expression of BAX and LC3b was significantly enhanced with CBD treatment when compared to control. LAMP1 and LC3b colocalization could also be observed with CBD and CBG treatment indicating changes in autophagic flux. A significant inhibition of migration, invasion and colony formation ability was shown in both CBD and CBG treatment in CCA. Western blot showed an overall decrease in the ratio of anti-apoptotic protein Bcl-2 with respect to pro-apoptotic protein BAX with CBG treatment. Furthermore, CBD treatment enhanced the expression of Type II cell death (autophagic degeneration) protein LC3b, which was reduced in CBG-treated CCA cells. Meanwhile, CBG treatment upregulated Type I cell death (programmed apoptosis) protein cleaved caspase-3. CBD and CBG are effective anti-cancer agents against CCA, capable of inhibiting the classic hallmarks of cancer, with a divergent mechanism of action (Type II or Type I respectively) in inducing these effects.


Dual roles for Prox1 in the regulation of the chicken betaB1-crystallin promoter.
Lens fiber cell differentiation is marked by the onset of betaB1-crystallin expression and is controlled by the cooperative action of a set of transcription factors including Prox1, an atypical homeodomain protein. Previously, the authors reported that Prox1 directly interacts with the OL2 element found in the chicken betaB1-crystallin basal promoter to activate the expression of this gene. Here they mapped the location of activating and repressing sequences of the full-length chicken betaB1-crystallin promoter (-432/+30) in lens epithelial cells, annular pad cells, and intact lens and characterized Prox1-binding sites found in this region. Transfection analysis and transgenic mice were used to characterize upstream regions of the chicken betaB1-crystallin gene. DNaseI footprinting and chromatin immunoprecipitation was performed to identify Prox1-binding sites, and transfection analyses were used to characterize these sites functionally. Sequences between -152 and -432 of the chicken betaB1-crystallin promoter mediated either promoter activation or repression, depending on the stage of lens differentiation tested. Two new Prox1-binding sites were found in this region that bound Prox1 more avidly than the OL2 element. However, neither binding site conferred Prox1-mediated activation on a heterologous promoter; instead, each allowed Prox1 to repress promoter function. The function of the upstream region of the chicken betaB1-crystallin promoter changes depending on cellular context. These data suggest that Prox1 function as a transcriptional activator could be regulated at the DNA level based on the characteristics of the responsive elements.


Geochemical Assessment of Trace Element Pollution in Surface Sediments from the Georges River, Southern Sydney, Australia.
Measurement of elevated trace elements is an important component of environmental assessment and management of estuarine marine sediments in systems adjacent to concentrated human activity. The present study surveys the estuarine sediments in selected tributary bays, creeks, and the upper segments of the Georges River system, NSW, Australia, which flows into the Tasman Sea through Botany Bay. A total of 146 surface sediment samples were analysed by X-ray fluorescence. Potential pollution of sediments was evaluated using potential load index, modified degree of contamination, and potential ecological risk index. The spatial distribution of trace elements varies between sites. Variable sources of contamination, including runoff from catchment areas, and emissions from watercraft and boatyards are contributing sources. Bay morphologies and their interactions with catchment and tidal flows play significant roles in the distribution of trace elements. The greatest concentration of trace elements occurs around discharge points and in the inner parts of bays that have high percentages of mud particles and organic matter. The lowest contamination by trace elements was found in sandy sediments along the shoreline and edges of the bays. Trace element distributions decline in concentration in residential-free areas and reach background levels in deeper sediment cores. The concentrations of trace elements were controlled by discharge points from the catchment area, marine boat activities, bay morphology, and sediment types (sand, silt, and clay). The highest pollutant concentrations are the result of past legal, but uncontrolled, discharge of waste from manufacturing into Salt Pan Creek.


EMR-Based Intervention Improves Lead Screening at an Urban Family Medicine Practice.
Elevated blood lead levels have well-described detrimental effects to growth and development in children, yet screening rates remain low. We sought to determine if a reminder within the electronic health record (EHR) could change provider behavior and improve blood lead level (BLL) screening test ordering rates in an urban academic family medicine practice. Baseline BLL test ordering rates were calculated for children ages 9-72 months. An update adding reminders to screen was made to the electronic note template used during pediatric well and sick visits at the practice. Data from the 10-week periods both before and after the change was made were compared through a retrospective chart review. A total of 210 children were seen during the pre-intervention period. Forty-eight percent (n=101) had already been screened. Of the 109 eligible for screening, 23 had tests ordered, and 18 of those had tests completed. Eighty-four children were eligible for screening in the post-intervention period. Forty-one of those children had tests ordered, and 15 had tests completed. Provider ordering rates increased from 21% of eligible patients to 49%. Test completion rates only increased from 17% to 18%. An electronic note-based reminder system significantly improves provider ordering rates of BLL tests. Researchers are currently investigating how the use of point-of-care BLL sample collection can improve test completion rates and therefore increase the frequency of successful screening.


A history of biological disasters of animal origin in North America.
This paper examines past occurrences in North America relevant to the possibility of biological disasters with animal origins. With respect to naturally occurring animal disease outbreaks, North America, while not as adversely affected by epizootics as other regions, has had its fair share of such outbreaks of both 'traditional' and emerging animal diseases. The traditional category includes such diseases as anthrax, classical swine fever, bluetongue, brucellosis, foot and mouth disease, and the family of equine encephalomyelitis viruses. The emerging diseases include relatively more recent culprits such as postweaning multisystemic wasting syndrome, poultry enteritis mortality syndrome, and newly discovered examples of the transmissible spongiform encephalopathies. Additionally, several serious diseases of human beings that involve animal vectors or reservoirs occur naturally in North America or have emerged in recent decades; these include plague, hantavirus, monkeypox, West Nile virus and avian-derived influenza. At the same time, there have been very few intentional attacks on livestock using biological agents and no recorded cases in North America of animals intentionally being used to transmit disease to humans. According to the historical record, therefore, naturally occurring emerging zoonoses probably constitute the greatest threat in terms of biological disasters with animal origins. However, some of the general trends in terrorist activity, such as the intensification of activities by animal rights extremists against facilities undertaking animal research, mean that the possibility of intentional animal-related biological disasters should not be discounted.


Propiconazole is a specific and accessible brassinosteroid (BR) biosynthesis inhibitor for Arabidopsis and maize.
Brassinosteroids (BRs) are steroidal hormones that play pivotal roles during plant development. In addition to the characterization of BR deficient mutants, specific BR biosynthesis inhibitors played an essential role in the elucidation of BR function in plants. However, high costs and limited availability of common BR biosynthetic inhibitors constrain their key advantage as a species-independent tool to investigate BR function. We studied propiconazole (Pcz) as an alternative to the BR inhibitor brassinazole (Brz). Arabidopsis seedlings treated with Pcz phenocopied BR biosynthetic mutants. The steady state mRNA levels of BR, but not gibberellic acid (GA), regulated genes increased proportional to the concentrations of Pcz. Moreover, root inhibition and Pcz-induced expression of BR biosynthetic genes were rescued by 24epi-brassinolide, but not by GA(3) co-applications. Maize seedlings treated with Pcz showed impaired mesocotyl, coleoptile, and true leaf elongation. Interestingly, the genetic background strongly impacted the tissue specific sensitivity towards Pcz. Based on these findings we conclude that Pcz is a potent and specific inhibitor of BR biosynthesis and an alternative to Brz. The reduced cost and increased availability of Pcz, compared to Brz, opens new possibilities to study BR function in larger crop species.


A comparative study of neuroprotective effect of angiotensin converting enzyme inhibitors against scopolamine-induced memory impairments in rats.
The comparative study of neuroprotective effect of angiotensin converting enzyme inhibitors against scopolamine-induced neuroinflammation in albino Wistar rats was studied. Male albino rats were administered with scopolamine to induce memory impairment. The standard nootropic agent, piracetam (200 mg/kg b.w., [i.p.]), perindopril (0.1 mg/kg b.w., [i.p.]), enalapril (0.1 mg/kg b.w., [i.p.]), and ramipril (0.1 mg/kg b.w., [i.p.]) were administered in different group of animals for 5 days. On 5(th) day, scopolamine (1 mg/kg b.w., i.p.) was administered after 60 min of the last dose of test drug. Memory function was evaluated in Morris water maze (MWM) test and pole climbing test (PCT). Biochemical estimations like glutathione (GSH), malondialdehyde (MDA), and acetylcholinesterase activity in the brain were estimated after completion of behavior study. All three test groups shows improvement in learning and memory in comparison to control group. Perindopril treated group showed a more effective significant decrease in escape latency time and transfer latency time compared to enalapril and ramipril treated group on day 4 in MWM test and PCT, respectively. Perindopril shows a significant reduction in MDA level and acetylcholinesterase activity and a significant rise in GSH level compared to enalapril and ramipril. The finding of this study indicates that Perindopril is more effective in memory retention compared to enalapril and ramipril.


A cell-based assay for aggregation inhibitors as therapeutics of polyglutamine-repeat disease and validation in Drosophila.
The formation of polyglutamine-containing aggregates and inclusions are hallmarks of pathogenesis in Huntington's disease that can be recapitulated in model systems. Although the contribution of inclusions to pathogenesis is unclear, cell-based assays can be used to screen for chemical compounds that affect aggregation and may provide therapeutic benefit. We have developed inducible PC12 cell-culture models to screen for loss of visible aggregates. To test the validity of this approach, compounds that inhibit aggregation in the PC12 cell-based screen were tested in a Drosophila model of polyglutamine-repeat disease. The disruption of aggregation in PC12 cells strongly correlates with suppression of neuronal degeneration in Drosophila. Thus, the engineered PC12 cells coupled with the Drosophila model provide a rapid and effective method to screen and validate compounds.


Electroless deposition and nanolithography can control the formation of materials at the nano-scale for plasmonic applications.
The new revolution in materials science is being driven by our ability to manipulate matter at the molecular level to create structures with novel functions and properties. The aim of this paper is to explore new strategies to obtain plasmonic metal nanostructures through the combination of a top down method, that is electron beam lithography, and a bottom up technique, that is the chemical electroless deposition. This technique allows a tight control over the shape and size of bi- and three-dimensional metal patterns at the nano scale. The resulting nanostructures can be used as constituents of Surface Enhanced Raman Spectroscopy (SERS) substrates, where the electromagnetic field is strongly amplified. Our results indicate that, in electroless growth, high quality metal nanostructures with sizes below 50 nm may be easily obtained. These findings were explained within the framework of a diffusion limited aggregation (DLA) model, that is a simulation model that makes it possible to decipher, at an atomic level, the rules governing the evolution of the growth front; moreover, we give a description of the physical mechanisms of growth at a basic level. In the discussion, we show how these findings can be utilized to fabricate dimers of silver nanospheres where the size and shape of those spheres is controlled with extreme precision and can be used for very large area SERS substrates and nano-optics, for single molecule detection.


Pest categorisation of Lasiodiplodia pseudotheobromae.
The EFSA Plant Health Panel performed a pest categorisation of Lasiodiplodia pseudotheobromae, a clearly defined fungus of the family Botryosphaeriaceae, which was first described in 2008 as a cryptic species within the L. theobromae complex. The pathogen affects a wide range of woody perennial crops and ornamental plants causing root rot, damping-off, leaf spots, twig blight, cankers, stem-end rot, gummosis, branch dieback and pre- and post-harvest fruit rots. Lasiodiplodia pseudotheobromae is present in Africa, Asia, North and South America and Oceania and has also been reported from Spain with a restricted distribution. However, there is uncertainty on the status of the pathogen worldwide and in the EU because in the past, when molecular tools (particularly multigene phylogenetic analysis) were not available, the pathogen might have been misidentified as L. theobromae. Lasiodiplodia pseudotheobromae is not included in Commission Implementing Regulation (EU) 2019/2072 and there are no interceptions in the EU. Because of the very wide host range of the pathogen, this pest categorisation focused on those hosts for which there is robust evidence that the pathogen was formally identified by a combination of morphology, pathogenicity and multilocus sequence analysis. Plants for planting, including seeds, fresh fruits and bark and wood of host plants as well as soil and other plant-growing media are the main pathways for the further entry of the pathogen into the EU. Host availability and climate suitability factors occurring in parts of the EU are favourable for the further establishment of the pathogen. In the area of its present distribution, including Spain, the pathogen has a direct impact on cultivated hosts. multilocus measures are available to prevent the further introduction and spread of the pathogen into the EU. Lasiodiplodia pseudotheobromae satisfies the criteria that are within the remit of EFSA to assess for this species to be regarded as potential Union quarantine pest.


Analysis of Trypanosoma brucei vsg expression site switching in vitro.
Trypanosoma brucei can undergo antigenic variation by switching between distinct telomeric variant surface glycoprotein gene (vsg) expression sites (ESs) or by replacing the active vsg. DNA rearrangements have often been associated with ES switching, but it is unclear if such rearrangements are necessary or whether ES inactivation always accompanies ES activation. To explore these issues, we derived ten independent clones, from the same parent, that had undergone a similar vsg activation event. This was achieved in the absence of an immune response, in vitro, using cells with selectable markers integrated into an ES. Nine of the ten clones had undergone ES switching. Such heritable changes in transcription state occurred at a frequency of approximately 6 x 10(-7). Comparison of switched and un-switched clones highlighted the dynamic nature of T. brucei telomeres, but changes in telomere length were not specifically associated with ES switching. Mapping within and beyond the ESs revealed no detectable DNA rearrangements, indicating that rearrangements are not necessary for ES activation/inactivation. Examination of individual cells indicated that ES activation consistently accompanied inactivation of the previously active ES. In some cases, however, we found cells that appeared to have efficiently established the switched state but which subsequently, at a frequency of approximately 2 x 10(-3), generated cells expressing both pre- and post-switch vsgs. These results show that ES activation/inactivation is usually a coupled process but that cells can inherit a propensity to uncouple these events.