Perception of obese schoolchildren regarding their participation in the Physical Education class and their level of self-esteem: comparison according to corporal status.
adequate exposure to Physical Education (PE) classes contributes to a healthier lifestyle among children during their lifespan. the main objective of this research was to evaluate the perception of obese schoolchildren regarding their participation in PE class and to determine their self-esteem compared with overweight and normal weight groups. in total, 656 schoolchildren (284 females and 372 males), between eleven and 14 years of age, participated in this study. Assessments were made regarding age, sex, anthropometric variables, children's self-esteem, and perception of participation in the PE class. RESULTS: obese schoolchildren presented significantly lower values of self-esteem (p < 0.001), and exhibited the highest proportion of schoolchildren with low self-esteem (53%). There was a significant relationship between obesity and the following sentence: "I am the last one they choose for games and in games and sports"; 48.53% of schoolchildren with obesity responded positively. With the following sentence: "I look instead of playing", 57.4% of schoolchildren with obesity responded positively. the results revealed that schoolchildren with obesity feel excluded from PE classes and show low levels of self-esteem, compared with normal weight schoolchildren.


Practical guidelines addressing ethical issues pertaining to the curation of human locus-specific variation databases (LSDBs).
More than 1,000 Web-based locus-specific variation databases (LSDBs) are listed on the Website of the Human Genetic Variation Society (HGVS). These individual efforts, which often relate phenotype to genotype, are a valuable source of information for clinicians, patients, and their families, as well as for basic research. The initiators of the Human Variome Project recently recognized that having access to some of the immense resources of unpublished information already present in diagnostic laboratories would provide critical data to help manage genetic disorders. However, there are significant ethical issues involved in sharing these data worldwide. An international working group presents second-generation guidelines addressing ethical issues relating to the curation of human LSDBs that provide information via a Web-based interface. It is intended that these should help current and future curators and may also inform the future decisions of ethics committees and legislators. These guidelines have been reviewed by the Ethics Committee of the Human Genome Organization (HUGO).


Mechanism responsible for inactivation of skeletal muscle pyruvate dehydrogenase complex in starvation and diabetes.
Regulation of the activity of the pyruvate dehydrogenase complex in skeletal muscle plays an important role in fuel selection and glucose homeostasis. Activation of the complex promotes disposal of glucose, whereas inactivation conserves substrates for hepatic glucose production. Starvation and diabetes induce a stable increase in pyruvate dehydrogenase kinase activity in skeletal muscle mitochondria that promotes phosphorylation and inactivation of the complex. The present study shows that these metabolic conditions induce a large increase in the expression of PDK4, one of four pyruvate dehydrogenase kinase isoenzymes expressed in mammalian tissues, in the mitochondria of gastrocnemius muscle. Refeeding starved rats and insulin treatment of diabetic rats decreased pyruvate dehydrogenase kinase activity and also reversed the increase in PDK4 protein in gastrocnemius muscle mitochondria. Starvation and diabetes also increased the abundance of PDK4 mRNA in gastrocnemius muscle, and refeeding and insulin treatment again reversed the effects of starvation and diabetes. These findings suggest that an increase in amount of this enzyme contributes to hyperphosphorylation and inactivation of the pyruvate dehydrogenase complex in these metabolic conditions. It was further found that feeding rats WY-14,643, a selective agonist for the peroxisome proliferator-activated receptor-alpha (PPAR-alpha), also induced large increases in pyruvate dehydrogenase kinase activity, PDK4 protein, and PDK4 mRNA in gastrocnemius muscle. Since long-chain fatty acids activate PPAR-alpha endogenously, increased levels of these compounds in starvation and diabetes may signal increased expression of PDK4 in skeletal muscle.


Spatial gradients of healthy aging: a study of myelin-sensitive maps.
Protracted development of a brain network may entail greater susceptibility to aging decline, supported by evidence of an earlier onset of age-related changes in late-maturing anterior areas, that is, an anterior-to-posterior gradient of brain aging. Here we analyzed the spatiotemporal features of age-related differences in myelin content across the human brain indexed by magnetization transfer (MT) concentration in a cross-sectional cohort of healthy adults. We described age-related spatial gradients in MT, which may reflect the reversal of patterns observed in development. We confirmed an anterior-to-posterior gradient of age-related MT decrease and also showed a lateral-to-ventral gradient inversely mirroring the sequence of connectivity development and myelination. MT concentration in the lateral white matter regions continued to increase up to the age of 45 years and decreased moderately following a peak. In contrast, ventral white matter regions reflected life-long stable MT concentration levels, followed by a rapid decrease at a later age. We discussed our findings in relation with existing theories of brain aging, including the lack of support for the proposal that areas which mature later decline at an accelerated rate.


Intracellular pH regulation in unstimulated Calliphora salivary glands is Na+ dependent and requires V-ATPase activity.
Salivary gland cells of the blowfly Calliphora vicina have a vacuolar-type H(+)-ATPase (V-ATPase) that lies in their apical membrane and energizes the secretion of a KCl-rich primary saliva upon stimulation with serotonin (5-hydroxytryptamine). Whether and to what extent V-ATPase contributes to intracellular pH (pH(i)) regulation in unstimulated gland cells is unknown. We used the fluorescent dye BCECF to study intracellular pH(i) regulation microfluorometrically and show that: (1) under resting conditions, the application of Na(+)-free physiological saline induces an intracellular alkalinization attributable to the inhibition of the activity of a Na(+)-dependent glutamate transporter; (2) the maintenance of resting pH(i) is Na(+), Cl(-), concanamycin A and DIDS sensitive; (3) recovery from an intracellular acid load is Na(+) sensitive and requires V-ATPase activity; (4) the Na(+)/H(+) antiporter is not involved in pH(i) recovery after a NH(4)Cl prepulse; and (5) at least one Na(+)-dependent transporter and the V-ATPase maintain recovery from an intracellular acid load. Thus, under resting conditions, the V-ATPase and at least one Na(+)-dependent transporter maintain normal pH(i) values of pH 7.5. We have also detected the presence of a Na(+)-dependent glutamate transporter, which seems to act as an acid loader. Despite this not being a common pH(i)-regulating transporter, its activity affects steady-state pH(i) in C. vicina salivary gland cells.


A prospective study of the factors affecting outcomes of nonsurgical root canal treatment: part 1: periapical health.
To investigate the probability of and factors influencing periapical status of teeth following primary (1°RCTx) or secondary (2°RCTx) root canal treatment. This prospective study involved annual clinical and radiographic follow-up of 1°RCTx (1170 roots, 702 teeth and 534 patients) or 2°RCTx (1314 roots, 750 teeth and 559 patients) carried out by Endodontic postgraduate students for 2-4 (50%) years. Pre-, intra- and postoperative data were collected prospectively on customized forms. The proportion of roots with complete periapical healing was estimated, and prognostic factors were investigated using multiple logistic regression models. Clustering effects within patients were adjusted in all models using robust standard error. proportion of roots with complete periapical healing after 1°RCTx (83%; 95% CI: 81%, 85%) or 2°RCTx (80%; 95% CI: 78%, 82%) were similar. Eleven prognostic factors were identified. The conditions that were found to improve periapical healing significantly were: the preoperative absence of a periapical lesion (P = 0.003); in presence of a periapical lesion, the smaller its size (P ≤ 0.001), the better the treatment prognosis; the absence of a preoperative sinus tract (P = 0.001); achievement of patency at the canal terminus (P = 0.001); extension of canal cleaning as close as possible to its apical terminus (P = 0.001); the use of ethylene-diamine-tetra-acetic acid (EDTA) solution as a penultimate wash followed by final rinse with NaOCl solution in 2°RCTx cases (P = 0.002); abstaining from using 2% chlorexidine as an adjunct irrigant to NaOCl solution (P = 0.01); absence of tooth/root perforation (P = 0.06); absence of interappointment flare-up (pain or swelling) (P =0.002); absence of root-filling extrusion (P ≤ 0.001); and presence of a satisfactory coronal restoration (P ≤ 0.001). Success based on periapical health associated with roots following 1°RCTx (83%) or 2°RCTx (80%) was similar, with 10 factors having a common effect on both, whilst the 11th factor 'EDTA as an additional irrigant' had different effects on the two treatments.


The use of pulse oximetry in evaluation of pulp vitality in immature permanent teeth.
The current methods of pulp vitality assessment, either electric or thermal, are of limited use in children. Recently, traumatized and immature teeth may not respond to such methods and because such methods require subjective responses, it may not provide accurate results particularly in children. Pulse oximetry, an atraumatic approach, is used to measure oxygen saturation in vascular system. The aim of this study was to investigate the use of pulse oximetry to evaluate pulp vitality status in immature permanent teeth. The study was conducted on 329 maxillary central and lateral incisors in children. The negative control group consisted of 10 root filled teeth. Systemic oxygen saturation was first measured on the thumb of the individual using a custom-made sensor. Oxygen saturation values of the teeth were then evaluated. The correlation between oxygen saturation measurement obtained from finger and tooth, and the correlation between oxygen saturation values and stage of root development were analyzed. A further comparison was made between the teeth with open and closed apex. Mean oxygen values recorded in the patient's finger were 97.17%, and mean oxygen values in the maxillary central and lateral incisors were 86.77% and 83/92%, respectively. There was no significant correlation between blood oxygen levels in the finger and in the teeth. (P > 0.05) There was a significant negative correlation between the stage of root development and the blood oxygen levels in the patients' teeth. (P < 0.05) Mean oxygen values in the teeth with open apex were significantly higher than the teeth with closed apex. (P < 0.001). Vital teeth provided consistent oxygen saturation readings, and non-vital teeth recorded no oxygen saturation values. During tooth development, the oxygen saturation values decreased. These findings confirm that the pulse oximetry is capable of detecting the pulpal blood flow and oxygen saturation.


A mixed methods assessment of knowledge, attitudes and practices related to aflatoxin contamination and exposure among caregivers of children under 5 years in western Kenya.
Identifying factors that may influence aflatoxin exposure in children under 5 years of age living in farming households in western Kenya. We used a mixed methods design. The quantitative component entailed serial cross-sectional interviews in 250 farming households to examine crop processing and conservation practices, household food storage and consumption and local understandings of aflatoxins. Qualitative data collection included focus group discussions (N 7) and key informant interviews (N 13) to explore explanations of harvesting and post-harvesting techniques and perceptions of crop spoilage. The study was carried out in Asembo, a rural community where high rates of child stunting exist. A total of 250 female primary caregivers of children under 5 years of age and thirteen experts in farming and food management participated. Study results showed that from a young age, children routinely ate maize-based dishes. Economic constraints and changing environmental patterns guided the application of sub-optimal crop practices involving early harvest, poor drying, mixing spoiled with good cereals and storing cereals in polypropylene bags in confined quarters occupied by humans and livestock and raising risks of aflatoxin contamination. Most (80 %) smallholder farmers were unaware of aflatoxins and their harmful economic and health consequences. Young children living in subsistence farming households may be at risk of exposure to aflatoxins and consequent ill health and stunting. Sustained efforts to increase awareness of the risks of aflatoxins and control measures among subsistence farmers could help to mitigate practices that raise exposure.


Resist diabetes: A randomized clinical trial for resistance training maintenance in adults with prediabetes.
To determine whether a social cognitive theory (SCT)-based intervention improves resistance training (RT) maintenance and strength, and reduces prediabetes prevalence. Sedentary, overweight/obese (BMI: 25-39.9 kg/m2) adults aged 50-69 (N = 170) with prediabetes participated in the 15-month trial. Participants completed a supervised 3-month RT (2×/wk) phase and were randomly assigned (N = 159) to one of two 6-month maintenance conditions: SCT or standard care. Participants continued RT at a self-selected facility. The final 6-month period involved no contact. Assessments occurred at baseline and months 3, 9, and 15. The SCT faded-contact intervention consisted of nine tailored transition (i.e., supervised training to training alone) and nine follow-up sessions. Standard care involved six generic follow-up sessions. Primary outcomes were prevalence of normoglycemia and muscular strength. The retention rate was 76%. Four serious adverse events were reported. After 3 months of RT, 34% of participants were no longer prediabetic. This prevalence of normoglycemia was maintained through month 15 (30%), with no group difference. There was an 18% increase in the odds of being normoglycemic for each % increase in fat-free mass. Increases in muscular strength were evident at month 3 and maintained through month 15 (P<0.001), which represented improvements of 21% and 14% for chest and leg press, respectively. Results did not demonstrate a greater reduction in prediabetes prevalence in the SCT condition. Resistance training is an effective, maintainable strategy for reducing prediabetes prevalence and increasing muscular strength. Future research which promotes RT initiation and maintenance in clinical and community settings is warranted. ClinicalTrials.gov NCT01112709.


Discovery of novel FabF ligands inspired by platensimycin by integrating structure-based design with diversity-oriented synthetic accessibility.
An approach for designing bioactive small molecules has been developed in which de novo structure-based ligand design (SBLD) was focused on regions of chemical space accessible using a diversity-oriented synthetic approach. The approach was exploited in the design and synthesis of a focused library of platensimycin analogues in which the complex bridged ring system was replaced with a series of alternative ring systems. The affinity of the resulting compounds for the C163Q mutant of FabF was determined using a WaterLOGSY competition binding assay. Several compounds had significantly improved affinity for the protein relative to a reference ligand. The integration of synthetic accessibility with ligand design enabled focus to be placed on synthetically-accessible regions of chemical space that were relevant to the target protein under investigation.


Oral and dental anomalies in purebred, brachycephalic Persian and Exotic cats.
OBJECTIVE To determine the prevalence of dental anomalies in brachycephalic cats from various geographic regions and analyze potential relationships with oral disease. DESIGN Prospective multicenter cross-sectional study. ANIMALS 50 purebred Persian (n = 42) and Exotic (8) cats. PROCEDURES Cats were anesthetized, and a complete dental examination, dental charting, 3-view oral photography, and full-mouth dental radiography were performed. RESULTS Malocclusions were observed in 36 (72%) cats. Crowding of the teeth was evident in 28 (56%) cats, with the incisors being most commonly affected. Malpositioned teeth were noted in 32 (64%) cats, with abnormal orientation being the most common anomaly followed by rotation and impaction. Numerical abnormalities were present in 38 (76%) cats, including 6 (12%) with hyperdontia and 32 (64%) with hypodontia. Periodontal disease was reported in 44 (88%) cats, and cats with periodontal disease were older than cats without. Tooth resorption was evident in 35 (70%) cats. Overall, 123 of 1,349 (8.7%) teeth had external inflammatory resorption and 82 (6.1%) had external replacement resorption. The premolar teeth were the teeth most commonly affected with inflammatory resorption, whereas the canine teeth were the teeth most commonly affected with replacement resorption. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that because of their brachycephaly, Persian and Exotic cats have unique oral and dental features that may predispose them to dental disease (eg, tooth resorption and periodontal disease). Knowledge of the particular dental anomalies common in brachycephalic cats could aid in early detection and mitigation of dental disease in these breeds.


Distribution and pathogenicity of Batrachochytrium dendrobatidis in boreal toads from the Grand Teton area of western Wyoming.
The pathogen Batrachochytrium dendrobatidis (Bd), which causes the skin disease chytridiomycosis, has been linked to amphibian population declines and extinctions worldwide. Bd has been implicated in recent declines of boreal toads, Bufo boreas boreas, in Colorado but populations of boreal toads in western Wyoming have high prevalence of Bd without suffering catastrophic mortality. In a field and laboratory study, we investigated the prevalence of Bd in boreal toads from the Grand Teton ecosystem (GRTE) in Wyoming and tested the pathogenicity of Bd to these toads in several environments. The pathogen was present in breeding adults at all 10 sites sampled, with a mean prevalence of 67%. In an experiment with juvenile toadlets housed individually in wet environments, 10(6) zoospores of Bd isolated from GRTE caused lethal disease in all Wyoming and Colorado animals within 35 days. Survival time was longer in toadlets from Wyoming than Colorado and in toadlets spending more time in dry sites. In a second trial involving Colorado toadlets exposed to 35% fewer Bd zoospores, infection peaked and subsided over 68 days with no lethal chytridiomycosis in any treatment. However, compared with drier aquaria with dry refuges, Bd infection intensity was 41% higher in more humid aquaria and 81% higher without dry refuges available. Our findings suggest that although widely infected in nature, Wyoming toads may escape chytridiomycosis due to a slight advantage in innate resistance or because their native habitat hinders Bd growth or provides more opportunities to reduce pathogen loads behaviorally than in Colorado.


Centromere fission, not telomere erosion, triggers chromosomal instability in human carcinomas.
The majority of sporadic carcinomas suffer from a kind of genetic instability in which chromosome number changes occur together with segmental defects. This means that changes involving intact chromosomes accompany breakage-induced alterations. Whereas the causes of aneuploidy are described in detail, the origins of chromosome breakage in sporadic carcinomas remain disputed. The three main pathways of chromosomal instability (CIN) proposed until now (random breakage, telomere fusion and centromere fission) are largely based on animal models and in vitro experiments, and recent studies revealed several discrepancies between animal models and human cancer. Here, we discuss how the experimental systems translate to human carcinomas and compare the theoretical breakage products to data from patient material and cancer cell lines. The majority of chromosomal defects in human carcinomas comprises pericentromeric breaks that are captured by healthy telomeres, and only a minor proportion of chromosome fusions can be attributed to telomere erosion or random breakage. Centromere fission, not telomere erosion, is therefore the most probably trigger of CIN and early carcinogenesis. Similar centromere-telomere fusions might drive a subset of congenital defects and evolutionary chromosome changes.


Out of Focus: Facial Feedback Manipulation Modulates Automatic Processing of Unattended Emotional Faces.
Facial expressions provide information about an individual's intentions and emotions and are thus an important medium for nonverbal communication. Theories of embodied cognition assume that facial mimicry and resulting facial feedback plays an important role in the perception of facial emotional expressions. Although behavioral and electrophysiological studies have confirmed the influence of facial feedback on the perception of facial emotional expressions, the influence of facial feedback on the automatic processing of such stimuli is largely unexplored. The automatic processing of unattended facial expressions can be investigated by visual expression-related MMN. The expression-related MMN reflects a differential ERP of automatic detection of emotional changes elicited by rarely presented facial expressions (deviants) among frequently presented facial expressions (standards). In this study, we investigated the impact of facial feedback on the automatic processing of facial expressions. For this purpose, participants (n = 19) performed a centrally presented visual detection task while neutral (standard), happy, and sad faces (deviants) were presented peripherally. During the task, facial feedback was manipulated by different pen holding conditions (holding the pen with teeth, lips, or nondominant hand). Our results indicate that automatic processing of facial expressions is influenced and thus dependent on the own facial feedback.


Invasiveness of primary brain tumors.
Primary malignant neoplasms of the nervous system differ from other types of malignancy in several ways. Clinical progression is due to local invasive growth, while metastases outside the skull are rare. The tumors show no sharp delimitation from the surrounding normal tissue. At the edge, an ill-defined area of invasive tumor cells, reacting glial cells and inflammatory cells is present. At the same time the primary brain tumors are biologically heterogeneous. In this review, a short survey of markers for malignancy in primary brain tumors is given, and some properties of importance for invasive behavior, are listed. These include different cellular enzymes, phagocytotic property, locomotive and proliferative characteristics. Studies of primary brain tumors in situ show invasive growth into the surrounding brain tissue, often followed by hemorrhage and necrosis. In addition spread of tumor cells takes place along preexisting intracranial structures. Recently, several systems for the study of brain tumor invasiveness in culture have been elaborated. Both experimental and human gliomas have been tested. The target tissues include organ culture of embryonic chick heart muscle, chorioallantoic membrane, fetal rat brain tissue and reconstructed vessel walls. It has been shown that glioma cells are able to split junctions between normal cells. They destroy and phagocytose the normal cells and penetrate the normal tissue. The use of brain tissue and reaggregated brain cell cultures as target for glioma cells in culture opens the possibility for an elucidation of invasiveness as one of the most important properties of malignancy in the nervous system.


Therapeutic potentials of propolis and pollen on biochemical changes in reproductive function of L-NAME induced hypertensive male rats.
Hypertension is described as increased blood pressure based on changed hemodynamics and associated with an increased oxidative damage to reproductive function. This work is to determine therapeutic and protective effects of apitherapy products (propolis and pollen) on reproductive functions of L-NAME-induced hypertensive male rats. Experimental animals were indiscriminately separated into four groups of seven rats in each group: (I) Control, (II) L-NAME, (III) L-NAME+ propolis and (IV) L-NAME+ pollen. At the end of the experimental applications, the rats were decapitated by anesthesia and biochemical analyzes were performed on the removed testicular tissues. The levels of TOS, NF-κB and MDA in the L-NAME group compared to control have increased (p < 0.05). The levels of these parameters in L-NAME+ propolis and L-NAME+ pollen groups compared to the L-NAME group have decreased (p < 0.05). TAS levels, PON1 and CAT activities were significantly decreased in testis tissue samples in the L-NAME-induced group (p < 0.05). However, these parameters were significantly lower in the L-NAME plus propolis and pollen groups (p < 0.05) compared with rats administered L-NAME alone (p < 0.05). NO level significantly reduced (p < 0.05) in L-NAME group compared with control group. There was no statistically significant changes in the NO level of the L-NAME+ propolis group compared with the L-NAME-treated group (p > 0.05). It has been determined that ethanolic extracts of propolis and pollen, which are natural bee products in the regulation of rising blood pressure. Propolis or pollen is thought to help regulate reproductive function by inhibiting the functioning of inflammatory pathways leading to hypertension.


Cost-effectiveness analysis and mortality impact estimation of scaling-up pregnancy test kits in Madagascar, Ethiopia and Malawi.
Cost-effective, innovative approaches are needed to accelerate progress towards ending preventable infant, child and maternal mortality. To inform policy decisions, we conducted a cost-effectiveness analysis of adding urine pregnancy test kits to the maternal and reproductive services package offered at the community level in Madagascar, Ethiopia and Malawi. We used a decision tree model to compare the intervention with the status quo for each country. We also completed single factor sensitivity analyses and Monte Carlo simulations with 10 000 iterations to generate the probability distribution of the estimates and uncertainty limits. Among a hypothetical cohort of 100 000 women of reproductive age, we estimate that over a 1-year period, the intervention would save 26, 35 and 48 lives in Madagascar, Ethiopia, and Malawi, respectively. The Incremental Cost Effectiveness Ratio (ICER) for the cost per life saved varies by country: $2311 [95% Uncertainty Interval (UI): $1699; $3454] in Madagascar; $2969 [UI: $2260; $5041] in Ethiopia and $1228 [UI: $918; $1777] in Malawi. This equates to an average cost per Disability Adjusted Life Year (DALY) averted of $36.28, $47.95 and $21.92, respectively. Based on WHO criteria and a comparison with other maternal, newborn, and child health interventions, we conclude that the addition of urine pregnancy tests to an existing community health worker maternal and reproductive services package is highly cost-effective in all three countries. To optimize uptake of family planning and antenatal care services and, in turn, accelerate the reduction of mortality and DALYs, decision makers and program planners should consider adding urine pregnancy tests to the community-level package of services.


Translating sexual assault prevention from a college campus to a United States military installation: piloting the know-your-power bystander social marketing campaign.
One population that shares both similar and different characteristics with traditional college-age students is the U.S. Military. Similarities include a high concentration of 18- to 26-year-olds dealing with new found independence, peer pressure, and the presence of social norms that support violence and hypermasculinity. Sexual violence is a major public health problem in the United States, and because of the similarities in the age group of college and military populations, the problems regarding sexual violence in both constituencies have been well-documented. In the current pilot study we seek to add to both current knowledge about and promising practices of translating prevention strategies from one target audience to another. We describe how we translated, administered, and evaluated a bystander intervention social marketing campaign focused on sexual assault prevention that had been found to significantly affect attitude change on a college campus for a U.S. Army installation in Europe. In addition to demonstrating the process of translating prevention strategies across target audiences, findings from this pilot study contribute to the evaluation data on the effectiveness of sexual violence prevention strategies implemented with members of the U.S. Military. From our analysis, we see that research participants indicate that the degree to which the images resonate with them and the familiarity of the context (i.e., social self-identification) significantly effect the participants' personal responsibility for reducing sexual assault, confidence in acting as a bystander, and reported engagement as a bystander.


Turn-on fluorescence detection of protein by molecularly imprinted hydrogels based on supramolecular assembly of peptide multi-functional blocks.
Synthetic receptors for biomacromolecules lack the supramolecular self-assembly behavior typical of biological systems. Here we propose a new method for the preparation of protein imprinted polymers based on the specific interaction of a peptide multi-functional block with a protein target. This peptide block contains a protein-binding peptide domain, a polymerizable moiety at the C-terminus and an environment-sensitive fluorescent molecule at the N-terminus. The method relies on a preliminary step consisting of peptide/protein supramolecular assembly, followed by copolymerization with the most common acrylate monomers (acrylamide, acrylic acid and bis-acrylamide) to produce a protein imprinted hydrogel polymer. Such a peptide block can function as an active assistant recognition element to improve affinity, and guarantees its effective polymerization at the protein/cavity interface, allowing for proper placement of a dye. As a proof of concept, we chose Bovine Serum Albumin (BSA) as the protein target and built the peptide block around a BSA binding dodecapeptide, with an allyl group as the polymerizable moiety and a dansyl molecule as the responsive dye. Compared to conventional approaches these hydrogels showed higher affinity (more than 45%) and imprinted sensitivity (about twenty fold) to the target, with a great BSA selectivity with respect to ovalbumin (α = 1.25) and lysozyme (α = 6.02). Upon protein binding, computational and experimental observations showed a blue shift of the emission peak (down to 440 nm) and an increase of fluorescence emission (twofold) and average lifetime (Δτ = 4.3 ns). Such an approach generates recognition cavities with controlled chemical information and represents an a priori method for self-responsive materials. Provided a specific peptide and minimal optimization conditions are used, such a method could be easily implemented for any protein target.


A calorie-restriction diet supplemented with fish oil and high-protein powder is associated with reduced severity of metabolic syndrome in obese women.
The prevalence of metabolic syndrome (MetS) and obesity has increased worldwide, as well as in Taiwan, particularly in women aged>40 years. The purpose of this study was to elucidate the effects of a calorie-restriction diet (CR) supplemented with protein and n-3 polyunsaturated fatty acids (PUFAs) on women with MetS. A total of 143 eligible female participants were recruited and assigned to four dietary interventions such as 1500-kcal CR, calorie-restriction meal-replacement diet (CRMR), calorie-restriction diet with fish oil supplementation (CRF) and calorie-restriction meal-replacement diet with fish oil supplementation (CRMRF). The changes in anthropometric measures, metabolic profiles, inflammatory response and the Z-score of severity of MetS were evaluated. Among 143 female MetS patients enrolled, 136 patients completed the 12-week study. After the 12-week dietary interventions, we observed reductions in body weight (BW), body mass index (BMI) and waist circumference (WC) in all groups. BMI and triglyceride (TG) levels decreased significantly in the CRMR, CRF and CRMRF groups, but not in the CR group. The homeostasis model assessment of insulin resistance (HOMA-IR) had significantly improved in all four groups, and the levels of interleukin-6 (IL-6) and C-reactive protein (CRP) had significantly decreased in the CRF and CRMRF groups. Following the interventions, the changes in waist circumference (WC), mean arterial pressure (MAP), fasting blood glucose (FBG), TGs, HOMA-IR, CRP and IL-6 significantly correlated with the reductions in Z-score of MetS severity. Our study results indicate that a calorie-restriction dietary intervention combined with various macronutrients can reduce the severity of MetS in women and increase recovery from MetS by almost twofold in comparison with a CR alone.


Arguments for zero tolerance of sexual contact between doctors and patients.
Some doctors do enter into sexual relationships with patients. These relationships can be damaging to the patient involved. One response available to both individual doctors and to disciplinary bodies is to prohibit sexual contact between doctors and patients ("zero tolerance"). This paper considers five ways of arguing for a zero tolerance policy. The first rests on an empirical claim that such contact is almost always harmful to the patient involved. The second is based on a "principles" approach while the third originates in "virtues" ethics. The fourth argues that zero tolerance is an "a priori" truth. These four attempt to establish that the behaviour is always wrong and ought, therefore, to be prohibited. The fifth argument is counterfactual. It claims a policy that allowed sexual contact would have unacceptable consequences. Given the responsibility of regulatory bodies to protect the public, zero tolerance is a natural policy to develop.


Prevalence of osteoporosis and fractures among women prescribed osteoporosis medication in five European countries: the POSSIBLE EU study.
European observational 1-year study assessed osteoporosis and fracture patterns in 3,402 postmenopausal women prescribed osteoporosis medication. Almost 40% of patients had a previous fracture, while 25% had neither fracture nor dual energy X-ray absorptiometry (DXA) diagnosis and were prescribed medication, probably due to other risk factors. This analysis assessed osteoporosis and fracture prevalence in postmenopausal women prescribed osteoporosis treatment in the Prospective Observational Study Investigating Bone Loss Experience in Europe(POSSIBLE EU). Women in this observational, multicenter 1-year study were categorized by fracture history and location at baseline. Baseline characteristics were analyzed according to no DXA and DXA diagnosis (osteoporosis or osteopenia). Fractures occurring during the 1-year follow-up period were recorded. Of the 3,402 women enrolled, 39% had a previous fracture, of whom 30% had ≥ 2 fractures. One thousand seven hundred and eighty-four (52%) patients had a DXA diagnosis (osteoporosis 68%, osteopenia 31%, and unknown 1%). Among the osteoporosis patients, 37% had a previous fracture (hip 2.9%, vertebral 8.8%, and non-hip, non-vertebral 25%) and 35% had fractures associated with major trauma. Of the 3,402 women, 1,476 (43%) had no DXA diagnosis; of these, 57% had no fracture (25% of all women). Risk factors varied across patients with and without DXA diagnosis. During the 1-year follow-up period, the fracture incidence in patients with or without a previous fracture at baseline was 4.7% and 1.6%, respectively. Almost 40% of patients prescribed osteoporosis medication had a previous fracture, highlighting a population with advanced disease. In contrast, 25% of patients had neither a previous fracture nor DXA diagnosis and were prescribed treatment, probably due to other risk factors. There is a need for continued improvement of disease management in European women.


Diagnostic and prognostic value of glycosyltransferase mRNA in glioblastoma multiforme patients.
Glioblastoma multiforme (GBM) is the most common and aggressive primary human brain tumour in adults with an average survival of 11 months. The 2-year survival is less than 10%, and only a small proportion of patients are alive at 3 years. Despite improved treatment strategies and aggressive therapy, the prognosis of GBM has changed little in past decades. Thus, any test that can reliably and rapidly diagnose the tumour and predict patient survival could be a valuable tool. Herein we report the use of quantitative real-time polymerase chain reaction (PCR) to quantify five glycosyltransferase transcripts in gliomas. Our results indicate that measuring GM1 synthase (beta-1,3 galactosyltransferase) mRNA may provide a useful method for segregating GBMs from other types of gliomas. In these studies, 97% of gliomas (36/37 tumours) below a threshold value had a diagnosis of GBM compared with 49% (52/106 tumours) above the threshold. More importantly, the increased expression of GD3 synthase mRNA in combination with decreased GalNAcT message correlated with increased survival in 79 GBM patients (proportional hazards model controlling for age, P = 0.02). These data were further corroborated by a data analysis from one of our previous studies on gangliosides of 80 GBMs, in which increased amounts of GM3 and GD3 (which accumulate in the absence of GalNAcT) correlated with a longer survival (P < 0.01). Thus, measuring GalNAcT and GD3 transcripts may provide a rapid method to assess prognosis in GBM patients. In summary, the data indicate that measuring glycosyltransferase mRNA levels by real-time PCR may be clinically useful for determining both diagnosis and prognosis in GBM patients.


[Intravascular ultrasound in cardiology].
Intravascular ultrasound (IVUS) is a clinically useful tool that provides cross-sectional images of the coronary arterial lumen and wall. Diagnostic applications of IVUS include the evaluation of ambiguous lesions on angiography particularly at the bifurcations. IVUS is also useful in the assessment of coronary vasculopathy in cardiac transplant patients or it can help to diagnose abnormalities such as syndrome X or coronary artery spasm. IVUS can optimize the performing of percutaneous coronary interventions, especially stent implantation. It represents as well an optimal tool for assessing regression of atherosclerosis. Three-dimensional reconstruction, elastography and imaging guide wires are some of the recent advances in the field of intravascular ultrasound.


Pharmacokinetic and Pharmacodynamic Comparison of Intravenous and Inhaled Caspofungin.
Background: Aspergillosis is a serious fungal lung infection caused by Aspergillus spp. and is often fatal in immunocompromised patients. Current antifungal drug treatment and delivery results in modest efficacy in these patients may be due to low drug distribution to the lung. A comparison of intravenous (IV) caspofungin and lung-targeted inhaled caspofungin was conducted in rats. The goal was to determine the concentrations of drug at the site of infection and systemic distribution that leads to toxicity. This was performed to understand the difference in the in vitro activity of caspofungin and modest in vivo efficacy. Methods: Caspofungin was delivered to rats through IV injection and nose-only inhalation. Each cohort received a single 2 mg/kg dose of drug. Plasma and tissue samples were analyzed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) and drug levels were compared. Results: The lung drug level was above the minimum effective concentration for 168 hours in the inhaled group but <24 hours in the IV cohort. The lung Cmax and area under curve (AUC) in the inhaled group was 20 times higher than in the IV group. Lung-targeted delivery doubled lung drug half-life compared with IV delivery. Systemic distribution to the liver and kidney was 45% lower for the inhaled cohort than the IV group of animals. Conclusions: Based on pharmacokinetic and pharmacodynamic indices, lung-targeted inhaled caspofungin is likely to provide an improved therapeutic benefit without any increase in systemic toxicities. Furthermore, inhaled delivery supports a weekly dosing regimen instead of daily IV dosing.


Gantenerumab for the treatment of Alzheimer's disease.
Alzheimer's disease is the leading cause of dementia in the elderly, and there is no disease-modifying therapy yet available. Immunotherapy directed against the β-amyloid peptide may be capable of slowing the rate of disease progression. Gantenerumab is the first fully human anti-β-amyloid monoclonal antibody. To review the efficacy and safety of immunotherapy drugs and in particular gantenerumab, we used the database MEDLINE. The primary literature on gantenerumab is reviewed in its entirety. We also reviewed the English-language, pre-clinical and clinical trials designed to evaluate the efficacy or/and safety of immunotherapy drugs, from 1999 through 2011. Other Alzheimer's disease-passive immunotherapeutics currently in development, according to www.clinicaltrials.gov, are also discussed. Gantenerumab appears capable of reducing the cerebral β-amyloid peptide burden in patients with Alzheimer's disease. Its ability to slow disease progression remains uncertain because no clinical data are available at present. The next step will be to investigate whether removal of brain amyloid translates into clinical benefit for patients at doses of gantenerumab that reduce brain amyloid and are well tolerated.


Comprehensive behavioral analysis of pituitary adenylate cyclase-activating polypeptide (PACAP) knockout mice.
Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide acting as a neurotransmitter, neuromodulator, or neurotrophic factor. PACAP is widely expressed throughout the brain and exerts its functions through the PACAP-specific receptor (PAC(1)). Recent studies reveal that genetic variants of the PACAP and PAC(1) genes are associated with mental disorders, and several behavioral abnormalities of PACAP knockout (KO) mice are reported. However, an insufficient number of backcrosses was made using PACAP KO mice on the C57BL/6J background due to their postnatal mortality. To elucidate the effects of PACAP on neuropsychiatric function, the PACAP gene was knocked out in F1 hybrid mice (C57BL/6J × 129SvEv) for appropriate control of the genetic background. The PACAP KO mice were then subjected to a behavioral test battery. PACAP deficiency had no significant effects on neurological screen. As shown previously, the mice exhibited significantly increased locomotor activity in a novel environment and abnormal anxiety-like behavior, while no obvious differences between genotypes were shown in home cage (HC) activity. In contrast to previous reports, the PACAP KO mice showed normal prepulse inhibition (PPI) and slightly decreased depression-like behavior. Previous study demonstrates that the social interaction (SI) in a resident-intruder test was decreased in PACAP KO mice. On the other hand, we showed that PACAP KO mice exhibited increased SI in Crawley's three-chamber social approach test, although PACAP KO had no significant impact on SI in a HC. PACAP KO mice also exhibited mild performance deficit in working memory in an eight-arm radial maze (RM) and the T-maze (TM), while they did not show any significant abnormalities in the left-right discrimination task in the TM. These results suggest that PACAP has an important role in the regulation of locomotor activity, social behavior, anxiety-like behavior and, potentially, working memory.


Barriers and facilitators to HIV testing and linkage to primary care: narratives of people with advanced HIV in the Southeast.
Persons with unrecognized HIV infection forgo timely clinical intervention and may unknowingly transmit HIV to partners. However, in the USA, unrecognized infection and late diagnosis are common. To understand barriers and facilitators to HIV testing and care, we conducted a qualitative study of 24 HIV infected persons attending a Southeastern HIV clinic who presented with clinically advanced illness. The primary barrier to HIV testing prior to diagnosis was perception of risk; consequently, most participants were diagnosed after the onset of clinical symptoms. While most patients were anxious to initiate care rapidly after diagnosis, some felt frustrated by the passive process of connecting to specialty care. The first visit with an HIV care provider was identified as critical in the coping process for many patients. Implications for the implementation of Centers for Disease Control and Prevention HIV routine screening guidelines are discussed.


Predictors of Continued Breastfeeding at One Year among Women Attending Primary Healthcare Centers in Qatar: A Cross-Sectional Study.
The number of babies in Qatar being exclusively breastfed is significantly lower than the global target set by the World Health Organization. The purpose of this study was to assess knowledge, attitude, and practice (KAP), selected barriers, and professional support as well as their association with continued breastfeeding at one year of age. A sample of Qatari and non-Qatari mothers (N = 195) who attended a well-baby clinic held at primary health care centers in Qatar completed a self-administered questionnaire. Descriptive analysis, the Pearson Chi-squared test, and logistic regression were performed. Around 42% of the mothers stopped breastfeeding when their child was aged between 0 and 11 months old. Mothers who had only one or female child stopped breastfeeding between the ages of 0 and 6 months (p = 0.025, 0.059). The more optimal the breastfeeding practices followed by the mothers, the older the age of the infant when they stopped breastfeeding (p = 0.001). The following factors were inversely associated with breastfeeding duration: the mother's perceptions that she "did not know how to breastfeed," or "wasn't making enough milk," and the need "to return to work/school", with p = 0.022, 0.004, and 0.022, respectively. These findings present factors that should be considered when planning for health education and promotion programs to prolong breastfeeding duration in Qatar.


[Effect of puerarin on liver injury in KKAy mice with type 2 diabetes mellitus].
To study the possible pathogenic mechanism of liver injury in type 2 diabetes mellitus (T2DM) and the intervening effect of puerarin on it. Mice with T2DM (KKAy) were randomly divided into two groups, the model group and the puerarin group. And the C57BL/J mice of the same age were set up as normal controls. They were sacrificed at 28 weeks old for observing serum fasting blood glucose (FBG), triglyceride (TG), total cholesterol (TC), alanine aminotransferase (ALT), aspartate aminotransferase (AST) by automatic biochemistry; liver cell apoptosis by flow cytometry; pathomorphology by electron microscope; and mRNA expressions of bcl-2 and bax genes by RT-PCR; as well as the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), Na(+)-K(+)-ATPase; and content of malondialdehyde (MDA) in liver tissue by spectrophotometer. In KKAy mice, blood levels of FBG, TG, TC, ALT, AST and liver cell apoptosis rate were higher; the bax mRNA expression was higher and bcl-2 mRNA was lower markedly; the activities of SOD, GSH-Px, Na(+)-K(+)-ATPase in liver tissue were lower, and MDA content was higher than those in the normal control significantly (all P <0.01). Besides, mitochondria swelling and damage were found in liver tissue. While in the puerarin group after treatment, all the above-mentioned changes were alleviated to some extent. Obvious liver injury emerges in KKAy mice. Puerarin shows a protective effect on the T2DM caused oxidative damage by way of up-regulating bcl-2 to inhibit oxidative stress, and improving the energy metabolic dysfunction in liver of mice.


Reduced port minimally invasive distal pancreatectomy: single-port laparoscopic versus robotic single-site plus one-port distal pancreatectomy.
Minimally invasive surgery appears to be developing in multiple directions, including single-port laparoscopic (SPL), single-port robotic, reduced port laparoscopic, or single-site plus one-port robotic approach. The aim of study was to compare the short-term perioperative variables and outcomes of patients undergoing reduced port minimally invasive distal pancreatectomy (DP) via the SPL, or robotic single-site plus one-port (RSS + 1) approach. The medical records of 35 patients were retrospectively reviewed, who underwent SPL-DP (n = 22) or RSS + 1 DP (n = 13) at Korea University Ansan Hospital and Yonsei University Severance Hospital. The mean operation time in SPL group was significantly higher than that of RSS + 1 group (281 vs 192, p = .001). The mean blood loss in SPL was significantly larger than that of RSS + 1 group (163 vs 12, p = .002). The mean length of free resection margin in SPL group was significantly longer than that of RSS + 1 group (2.1 vs 0.4 cm, p = .001). Spleen was significantly preserved in SPL group (54.5 vs 7.7%, p = .001). All RSS + 1 cases had tumors located near spleen hilum (p < .001). SPL approach had significantly grade IIIa complications (p = .014). Moreover, the mean hospital stay in SPL group was significantly longer than that of RSS + 1 group (14.4 vs 7.4 days, p = .004). Postoperative pancreatic fistula (POPF) was significantly observed in longer operation time (p = .043) and smaller tumor size (p = .037) in the univariate analysis. Higher BMI was significantly important factor for prolonged operation time (p = .034) in the multivariate analysis. Prolonged hospital stay was related to spleen preservation (p = .014) in the multivariate analysis. Both SPL and RSS + 1 are technically feasible and safe. RSS + 1-DP is superior to SPL-DP in terms of operation time, blood loss, severe complications, and hospital stay. SPL-DP shows advantages in terms of single wound site, less trocar usage, higher rate of spleen preservation, and wider range of operative field.


High-throughput method for rapid induction of prophages from lysogens and its application in the study of Shiga Toxin-encoding Escherichia coli strains.
A high-throughput 96-well plate-based method for the rapid induction of endogenous prophages from individual bacterial strains was developed. The detection of endogenous prophages was achieved by the filtration of the culture liquor following norfloxacin induction and subsequent PCRs targeting bacteriophage-carried gene markers. The induction method was tested on 188 putative Shiga toxin (Stx)-producing Escherichia coli (STEC) strains and demonstrated the ability to detect both lambdoid and stx-carrying bacteriophages in strains for which plaques were not observed via plaque assay. Lambdoid bacteriophages were detected in 37% of the induced phage preparations via amplification of the Q gene, and Stx1- and Stx2-encoding phages were detected in 2 and 14% of the strains, respectively. The method therefore provided greater sensitivity for the detection of Stx and other lambdoid bacteriophage populations carried by STEC strains than that for the established method of plaque assay using bacterial indicator strains, enabling, for the first time, large-scale bacteriophage population and diversity studies.


High-resolution Fresnel zone plate fabrication by achromatic spatial frequency multiplication with extreme ultraviolet radiation.
We used an approach based on the self-imaging property of gratings to fabricate high-resolution Fresnel zone plates (FZPs). Under certain conditions, the illumination of a parent ZP with a wideband EUV beam produces a radially oscillating intensity distribution with double the spatial frequency of the ZP. This intensity distribution is observed in a certain distance range, given by the local zone width, the focal length of the ZP, and the spectral bandwidth of the illuminating beam. This phenomenon has been used to lithographically record daughter ZPs that have approximately half the zone width, thus twice the resolution, of the parent ZP. FZPs with zone widths as low as 30 nm have been fabricated in this way. Use of this technique in the extreme ultraviolet (EUV) region has the potential for high throughput production of FZPs and similar high-resolution diffraction optics with variable spatial frequency for the EUV and x-ray regions.


Differential rate of recovery in athletes after first and second concussion episodes.
Clinical observations suggest that a history of previous concussions may cause a slower recovery of neurological function after recurrent concussion episodes. However, direct examination of this notion has not been provided. This report investigates the differential rate of restoring the visual-kinesthetic integration in collegiate athletes experiencing single versus recurrent concussion episodes. One hundred sixty collegiate athletes were tested preseason using multimodal research methodology. Of these, 38 experienced mild traumatic brain injury (MTBI) and were tested on Days 10, 15, and 30 after injury. Nine of these MTBI patients experienced a second MTBI within 1 year after the first brain injury and were retested. The postconcussion symptoms checklist, neuropsychological evaluations, and postural responses to visual field motion were recorded using a virtual reality environment. All patients were asymptomatic at Day 10 of testing and were cleared for sport participation based on clinical symptoms resolution. Balance deficits, as evident by incoherence with visual field motion postural responses, were present at least 30 days after injury (P < 0.001). Most importantly, the rate of balance symptoms restoration was significantly reduced after a recurrent, second concussion (P < 0.001) compared with those after the first concussion. The findings of this study confirm our previous research indicating the presence of long-term residual visual-motor disintegration in concussed individuals with normal neuropsychological measures. Most importantly, athletes with a history of previous concussion demonstrate significantly slower rates of recovery of neurological functions after the second episode of MTBI.


Postexercise net protein synthesis in human muscle from orally administered amino acids.
We examined the response of net muscle protein synthesis to ingestion of amino acids after a bout of resistance exercise. A primed, constant infusion of L-[ring-2H5]phenylalanine was used to measure net muscle protein balance in three male and three female volunteers on three occasions. Subjects consumed in random order 1 liter of 1) a mixed amino acid (40 g) solution (MAA), 2) an essential amino acid (40 g) solution (EAA), and 3) a placebo solution (PLA). Arterial amino acid concentrations increased approximately 150-640% above baseline during ingestion of MAA and EAA. Net muscle protein balance was significantly increased from negative during PLA ingestion (-50 +/- 23 nmol. min-1. 100 ml leg volume-1) to positive during MAA ingestion (17 +/- 13 nmol. min-1. 100 ml leg volume-1) and EAA (29 +/- 14 nmol. min-1. 100 ml leg volume-1; P < 0.05). Because net balance was similar for MAA and EAA, it does not appear necessary to include nonessential amino acids in a formulation designed to elicit an anabolic response from muscle after exercise. We concluded that ingestion of oral essential amino acids results in a change from net muscle protein degradation to net muscle protein synthesis after heavy resistance exercise in humans similar to that seen when the amino acids were infused.


Intraoperative Nerve Monitoring in Thyroidectomies for Malignancy: Does It Matter?
Recurrent laryngeal nerve (RLN) injury and postoperative hypocalcemia are potential complications of thyroidectomy, particularly in malignancy. Intraoperative nerve monitoring (IONM) remains controversial. We sought to evaluate the impact of IONM on these complications using a national data set. The American College of Surgeons National Surgical Quality Improvement Program thyroidectomy-targeted data set was queried for patients who underwent thyroidectomies from 2016 to 2017. Patients were grouped according to IONM use. Logistic regression models were constructed to evaluate associations of variables with 30-day hypocalcemic events (HCEs) and RLN injury. Associations were expressed as odds ratios (ORs) with 95% confidence intervals (95% CIs). A subgroup analysis was performed of patients with malignancy. A total of 9527 patients were identified; 5969 (62.7%) underwent thyroidectomy with IONM and 3558 (37.3%) without. By multivariable analysis, IONM had protective associations with HCE (OR = .81, 95% CI = .68-.96; P = .013) and RLN injury (OR = .83, 95% CI = .69-.98; P = .033). Malignancy increased risk of HCE (OR = 1.21, 95% CI=1.01-1.45; P = .038) and RLN injury (OR = 1.22, 95% CI = 1.02-1.46; P = .034). A large proportion (5943/9527, 62.4%) of patients had malignancy; 3646 (61.3%) underwent thyroidectomy with IONM and 2297 (38.7%) without. In the subgroup analysis, IONM had stronger protective associations with HCE (OR = .73, 95% CI = .60-.90; P = .003) and RLN injury (OR = .76, 95% CI = .62-.94; P = .012). Malignancy was associated with increased risk of HCE and RLN injury. Intraoperative nerve monitoring had a protective association with HCE and RLN injury, both overall, and in the malignant subgroup. Intraoperative nerve monitoring was correlated with improved thyroidectomy outcomes, especially if the indication was malignancy. This warrants further study to clarify cause and effect.


Evaluation of a more ventral starting point for thoracic pedicle screws: higher maximal insertional arc and more medial and safer screw angulation.
OBJECTIVETo demonstrate that a more ventral starting point for thoracic pedicle screw insertion, produced by aggressively removing the dorsal transverse process bone down to the superior articular facet (SAF), results in a larger margin for error and more medial screw angulation compared to the traditional dorsal starting point (DSP). The margin for error will be quantified by the maximal insertional arc (MIA).METHODSThe study population included 10 consecutive operative patients with adult idiopathic scoliosis who underwent primary surgery. All measurements were performed using 3D visualization software by an attending spine surgeon. The screw starting points were 2 mm lateral to the midline of the SAF in the mediolateral direction and in the center of the pedicle in the cephalocaudal direction. The DSP was on the dorsal cortex. The ventral starting point (VSP) was at the depth of the SAF. Measurements included distance to the pedicle isthmus, MIA, and screw trajectories.RESULTSTen patients and 110 vertebral levels (T1-11) were measured. The patients' average age was 41.4 years (range 18-64 years). The pedicle isthmus was largest at T1 (4.04 ± 1.09 mm), and smallest at T5 (1.05 ± 0.93 mm). The distance to the pedicle isthmus was 7.47 mm for the VSP and 11.92 mm for the DSP (p < 0.001). The MIA was 15.3° for the VSP and 10.1° for the DSP (p < 0.001). Screw angulation was 21.7° for the VSP and 16.8° for the DSP (p < 0.001).CONCLUSIONSA more ventral starting point for thoracic pedicle screws results in increased MIA and more medial screw angulation. The increased MIA represents an increased tolerance for error that should improve the safety of pedicle screw placement. More medial screw angulation allows improved triangulation of pedicle screws.


Airflow and symptom outcomes between allergic and non-allergic rhinitis patients from turbinoplasty.
Inferior turbinate procedures are applied to relieve medically refractory nasal obstruction. However, the nature of congestion differs between allergic(AR) and non-allergic rhinitis(NAR). This study compares surgical outcomes between AR and NAR patients. A case-control study of patients undergoing turbinate with or without septoplasty surgery for nasal obstruction was performed. Patient reported outcomes were: nasal obstruction, global nasal function(GNF), and sino-nasal outcome test(SNOT-22) with rhinitis, facial symptom, sleep and psychological sub-scores. Nasal peak inspiratory flow(NPIF) assessed nasal airflow. Measurements were obtained preoperatively and 3 months postoperatively. 190 patients were assessed. AR had worse obstruction and worse GNF. All outcomes improved post-surgery; nasal obstruction, GNF, SNOT-22, rhinitis-symptoms, facial-symptoms, sleep-function, psychological-function and NPIF. GNF improvement was greater in AR. NPIF improvement was similar between groups. Both AR and NAR patients gained benefit from surgery to relieve nasal obstruction. AR patients demonstrate greater improvement in GNF score but allergy management may contribute to this.


Sputum eosinophilia and the short term response to inhaled mometasone in chronic obstructive pulmonary disease.
An association between the sputum eosinophil count and the response to a 2 week course of prednisolone has previously been reported in patients with chronic obstructive pulmonary disease (COPD). Whether the response to inhaled corticosteroids is related to the presence of eosinophilic inflammation is unclear. A randomised, double blind, crossover trial of placebo and mometasone furoate (800 microg/day), each given for 6 weeks with a 4 week washout period, was performed in subjects with COPD treated with bronchodilator therapy only. Spirometric tests, symptom scores, chronic respiratory disease questionnaire (CRQ), and induced sputum were performed before and after each treatment phase. Ninety five patients were recruited of which 60 were randomised. Overall there were no treatment associated changes in forced expiratory volume in 1 second (FEV(1)), total CRQ, or sputum characteristics. After stratification into tertiles by baseline eosinophil count, the net improvement in post-bronchodilator FEV(1) increased with mometasone compared with placebo progressively from the least to the most eosinophilic tertile. The mean change in post-bronchodilator FEV(1) with mometasone compared with placebo in the highest tertile was 0.11 l (95% CI 0.03 to 0.19). This improvement was not associated with a fall in the sputum eosinophil count. An increased sputum eosinophil count is related to an improvement in post-bronchodilator FEV(1) following treatment with inhaled mometasone in COPD, but the improvement is not associated with a reduction in the sputum eosinophil count.


Correction factors for assessing immersion suits under harsh conditions.
Many immersion suit standards require testing of thermal protective properties in calm, circulating water while these suits are typically used in harsher environments where they often underperform. Yet it can be expensive and logistically challenging to test immersion suits in realistic conditions. The goal of this work was to develop a set of correction factors that would allow suits to be tested in calm water yet ensure they will offer sufficient protection in harsher conditions. Two immersion studies, one dry and the other with 500 mL of water within the suit, were conducted in wind and waves to measure the change in suit insulation. In both studies, wind and waves resulted in a significantly lower immersed insulation value compared to calm water. The minimum required thermal insulation for maintaining heat balance can be calculated for a given mean skin temperature, metabolic heat production, and water temperature. Combining the physiological limits of sustainable cold water immersion and actual suit insulation, correction factors can be deduced for harsh conditions compared to calm. The minimum in-situ suit insulation to maintain thermal balance is 1.553-0.0624·TW + 0.00018·TW(2) for a dry calm condition. Multiplicative correction factors to the above equation are 1.37, 1.25, and 1.72 for wind + waves, 500 mL suit wetness, and both combined, respectively. Calm water certification tests of suit insulation should meet or exceed the minimum in-situ requirements to maintain thermal balance, and correction factors should be applied for a more realistic determination of minimum insulation for harsh conditions.


Situational assessment of hospital facilities for modernization purposes and resilience improvement.
Modernization of hospital facilities is one of the objectives of administrators and decision-makers of healthcare systems. Hospital facilities are both complex and critical infrastructures, because they are characterized by high level of interconnections, dynamism, technological innovation, and because they offer health and social essential services. Decision-makers have to implement modernization strategies of hospital facilities in order to guarantee a high standard of care and a resilient response during disasters and emergencies. The critical role played by hospital facilities is acknowledge by the international action programs, including the 2030 Agenda of United Nations for Sustainable Development, and it has been emphasized by the COVID-19 pandemic. The paper illustrates the RADAR-Hospital Facilities methodology (RADAR-HF) developed for the situational assessment of the physical environment of hospital facilities. RADAR-HF provides the decision-makers with an overview of the main aspects for modernization (safety, functionality, sustainability, adaptability, comfort) and substantial information for planning interventions, considering hospital facilities as interconnected systems. The outcomes are represented by ad-hoc designed graphical indicators and overview-tools, that summarize the status-conditions of one or a set of existing hospital facilities, the upgrading needs, and the best occupancy of facilities. Decision-makers could use RADAR-HF to define integrated modernization strategies with resilience improvement, monitor the situation of the facilities, and understand the effectiveness of interventions. The paper ends showing the results obtained in a research project, in which RADAR-HF has been applied to assess the existing hospital facilities of the Friuli Venezia Giulia region (North-East of Italy).


Specific and stable fluorescence labeling of histidine-tagged proteins for dissecting multi-protein complex formation.
Labeling of proteins with fluorescent dyes offers powerful means for monitoring protein interactions in vitro and in live cells. Only a few techniques for noncovalent fluorescence labeling with well-defined localization of the attached dye are currently available. Here, we present an efficient method for site-specific and stable noncovalent fluorescence labeling of histidine-tagged proteins. Different fluorophores were conjugated to a chemical recognition unit bearing three NTA moieties (tris-NTA). In contrast to the transient binding of conventional mono-NTA, the multivalent interaction of tris-NTA conjugated fluorophores with oligohistidine-tagged proteins resulted in complex lifetimes of more than an hour. The high selectivity of tris-NTA toward cumulated histidines enabled selective labeling of proteins in cell lysates and on the surface of live cells. Fluorescence labeling by tris-NTA conjugates was applied for the analysis of a ternary protein complex in solution and on surfaces. Formation of the complex and its stoichiometry was studied by analytical size exclusion chromatography and fluorescence quenching. The individual interactions were dissected on solid supports by using simultaneous mass-sensitive and multicolor fluorescence detection. Using these techniques, formation of a 1:1:1 stoichiometry by independent interactions of the receptor subunits with the ligand was shown. The incorporation of transition metal ions into the labeled proteins upon labeling with tris-NTA fluorophore conjugates provided an additional sensitive spectroscopic reporter for detecting and monitoring protein-protein interactions in real time. A broad application of these fluorescence conjugates for protein interaction analysis can be envisaged.


A highly selective telomerase inhibitor limiting human cancer cell proliferation.
Telomerase, the ribonucleoprotein enzyme maintaining the telomeres of eukaryotic chromosomes, is active in most human cancers and in germline cells but, with few exceptions, not in normal human somatic tissues. Telomere maintenance is essential to the replicative potential of malignant cells and the inhibition of telomerase can lead to telomere shortening and cessation of unrestrained proliferation. We describe novel chemical compounds which selectively inhibit telomerase in vitro and in vivo. Treatment of cancer cells with these inhibitors leads to progressive telomere shortening, with no acute cytotoxicity, but a proliferation arrest after a characteristic lag period with hallmarks of senescence, including morphological, mitotic and chromosomal aberrations and altered patterns of gene expression. Telomerase inhibition and telomere shortening also result in a marked reduction of the tumorigenic potential of drug-treated tumour cells in a mouse xenograft model. This model was also used to demonstrate in vivo efficacy with no adverse side effects and uncomplicated oral administration of the inhibitor. These findings indicate that potent and selective, non-nucleosidic telomerase inhibitors can be designed as novel cancer treatment modalities.


An evaluation of the predictive ability of the QSAR software packages, DEREK, HAZARDEXPERT and TOPKAT, to describe chemically-induced skin irritation.
According to the REACH chemicals legislation, formally adopted by the EU in 2006, Quantitative Structure-Activity Relationships (QSARs) can be used as alternatives to animal testing, which itself poses specific ethical and economical concerns. A critical assessment of the performance of the QSAR models is therefore the first step toward the reliable use of such computational techniques. This article reports the performance of the skin irritation module of three commercially-available software packages: DEREK, HAZARDEXPERT and TOPKAT. Their performances were tested on the basis of data published in the literature, for 116 chemicals. The results of this study show that only TOPKAT was able to predict the irritative potential for the majority of chemicals, whereas DEREK and HAZARDEXPERT could correctly identify only a few irritant substances.


Influence of metals on cytokines production in connection with successful implantation therapy in dentistry.
In most of patients in need of implantation treatment in the oral cavity, implants heal well, nevertheless, there are some individuals, in whom titanium implants fail for reasons, which remain unclear. The aim of our study was to determine if there is a difference between metal influenced IL-1β, IL-4, IL-6, TNF-α and IFN-γ cytokines production in patients with successfully healed implants compared to those, whose implant therapy was unsuccessful. The two study groups included 12 patients with failed dental titanium implants and 9 patients with successfully healed implants. In the subjects, cytokine production was established after lymphocyte cultivation with mercury, nickel and titanium antigens. IL-1β levels were significantly increased in all patients after stimulation with titanium and in patients with accepted implants compared to patients with failed implants after the stimulation with mercury and titanium. Titanium caused significantly increased IL-6 production in all patients. TNF-α and IFN-γ levels were also significantly increased after the stimulation with titanium. Significantly increased TNF-α levels were found in patients with accepted implants as compared to patients with failed implants. Increased production of IL-1β a IL-6 cytokines in reaction to titanium and increased production of TNF-α and IFN-γ cytokines in reaction to mercury, which is very often present in the form of amalgam in the oral cavity of persons in need of implant therapy, can play an important role in immune reactions during implant healing process. In patients with failed titanium implants, decreased production of these cytokines may participate in implant failure.


Cellular aging in depression: Permanent imprint or reversible process?: An overview of the current evidence, mechanistic pathways, and targets for interventions.
Depression might be associated with accelerated cellular aging. However, does this result in an irreversible state or is the body able to slow down or recover from such a process? Telomeres are DNA-protein complexes that protect the ends of chromosomes and generally shorten with age; and therefore index cellular aging. The majority of studies indicate that persons with depression have shorter leukocyte telomeres than similarly aged non-depressed persons, which may contribute to the observed unfavorable somatic health outcomes in the depressed population. Some small-scale preliminary studies raise the possibility that behavioral or pharmacological interventions may either slow down or else reverse this accelerated telomere shortening, possibly through increasing the activity of the telomere-lengthening enzyme telomerase. This paper covers the current state of evidence in the relationship between depression and the telomere-telomerase system and debates whether depression-related cellular aging should be considered a reversible process or permanent damage.


Flavanols and cardiovascular health: effects on the circulating NO pool in humans.
Atherosclerosis is the major cause for chronic vascular diseases. The key event in the pathogenesis of atherosclerosis is believed to be dysfunction of the endothelium and disruption of endothelial homeostasis, leading to vasoconstriction, inflammation, leukocyte adhesion, thrombosis, and proliferation of vascular smooth muscle cells. Endothelium-derived nitric oxide (NO) plays a major role in vascular homeostasis and a decrease in NO-bioavailability accelerates the development of atherosclerosis. Given that endothelial dysfunction is at least in part reversible, the characterization of endothelial function and therapeutical approaches have gained much attention over the past years. Recent studies demonstrated that especially the consumption of plant-derived foods rich in certain flavonoids can improve endothelial function in both compromised and healthy humans. Furthermore, various physiologic and biochemical measures have been used previously as biomarkers for the assessment of the proposed beneficial effects of flavonoids in this context. More recently, the analysis of plasma nitros(yl)ated species (RXNOs), referred to as the circulating NO pool, has gained recognition, especially as a marker for endothelial function. This review is aimed at evaluating the suitability of quantifying this NO pool as a biomarker for cardiovascular function in humans, in particular during dietary interventions with flavonoid-rich foods.


RNA G-quadruplexes cause eIF4A-dependent oncogene translation in cancer.
The translational control of oncoprotein expression is implicated in many cancers. Here we report an eIF4A RNA helicase-dependent mechanism of translational control that contributes to oncogenesis and underlies the anticancer effects of silvestrol and related compounds. For example, eIF4A promotes T-cell acute lymphoblastic leukaemia development in vivo and is required for leukaemia maintenance. Accordingly, inhibition of eIF4A with silvestrol has powerful therapeutic effects against murine and human leukaemic cells in vitro and in vivo. We use transcriptome-scale ribosome footprinting to identify the hallmarks of eIF4A-dependent transcripts. These include 5' untranslated region (UTR) sequences such as the 12-nucleotide guanine quartet (CGG)4 motif that can form RNA G-quadruplex structures. Notably, among the most eIF4A-dependent and silvestrol-sensitive transcripts are a number of oncogenes, superenhancer-associated transcription factors, and epigenetic regulators. Hence, the 5' UTRs of select cancer genes harbour a targetable requirement for the eIF4A RNA helicase.


Incorporating sprouted chickpea flour in pasta increases brachial artery flow-mediated dilation.
Consumption of alternative flours, such as sprouted chickpea flour, has shown increased popularity in recent years. Foods rich in antioxidants have been shown to influence brachial artery flow-mediated dilation (FMD), a non-invasive test of a crucial layer of the artery called the endothelium. Partially replacing the semolina flour in pasta with sprouted chickpea flour (SCF) may acutely affect endothelial function post-digestion. We sought to determine if FMD was higher, lower, or the same post-digestion of pasta made with 60% semolina flour and 40% SCF (SCF40) vs. post-digestion of pasta made with 100% semolina flour (SEM100, i.e., control). Trolox equivalent antioxidant capacity (TEAC) analysis was performed on the same flour samples. Healthy participants underwent a screening visit and two randomized controlled meal data collection visits (SCF40 and SEM100). At each data collection visit, participants consumed 255 g of pasta with butter. FMD was assessed 2-3 h after pasta consumption. TEAC results showed that SCF40 (2.031 ± 0.096 mmol trolox/100 g sample) had significantly greater antioxidant capacity than SEM100 (1.736 ± 0.046 mmol trolox/100 g sample; p = 0.02). Twenty-two healthy participants (5 men and 17 women; 26 ± 2 years, 66.6 ± 2.3 kg, BMI = 24 ± 1 kg/m2, SBP = 114 ± 3 mmHg, DBP = 75 ± 2 mmHg, HR = 74 ± 3 BPM) were studied. FMD in the SCF40 condition (10.3% ± 1.2%) was greater than the SEM100 condition (7.9% ± 0.8%, p = 0.02). These data suggest that partial substitution with sprouted chickpea flour in place of semolina flour in pasta acutely improves post-digestion FMD, which may be beneficial for cardiovascular health (ClinicalTrials.gov Identifier: NCT03801486).


Surface-initiated self-healing of polymers in aqueous media.
Polymeric materials that intrinsically heal at damage sites under wet or moist conditions are urgently needed for biomedical and environmental applications. Although hydrogels with self-mending properties have been engineered by means of mussel-inspired metal-chelating catechol-functionalized polymer networks, biological self-healing in wet conditions, as occurs in self-assembled holdfast proteins in mussels and other marine organisms, is generally thought to involve more than reversible metal chelates. Here we demonstrate self-mending in metal-free water of synthetic polyacrylate and polymethacrylate materials that are surface-functionalized with mussel-inspired catechols. Wet self-mending of scission in these polymers is initiated and accelerated by hydrogen bonding between interfacial catechol moieties, and consolidated by the recruitment of other non-covalent interactions contributed by subsurface moieties. The repaired and pristine samples show similar mechanical properties, suggesting that the triggering of complete self-healing is enabled underwater by the formation of extensive catechol-mediated interfacial hydrogen bonds.


Immunodominance of cytotoxic T lymphocyte epitopes co-injected in vivo and modulation by interleukin-12.
Immunodominance (ID) of T cell epitopes is a well-documented phenomenon that might have profound significance in the evolution of T cell responses to pathogens, tumors, autoantigens and vaccines. With the intention of developing vaccines composed of several cytotoxic T cell (CTL) epitopes, we injected mice with peptide mixtures containing two to five CTL epitopes and observed clear patterns of ID. In a first case, ID strictly correlated with the competitor activity of the individual peptides for H-2Kd, whereas in a second case, the absence of correlation between ID and competitor activity, binding affinity, half-life of the peptides in serum, induction of proliferation in vitro and the individual immunogenicity of the peptides, suggested to us that ID of co-injected CTL epitopes can be determined both at the peptide level (binding affinity to H-2Kd) and at the T cell level. This hypothesis is supported by our finding that interleukin-12 strongly modulates ID when it is not correlated with MHC binding.


Intra-ovarian roles of activins and inhibins.
Granulosa cells are the main ovarian source of inhibins, activins and activin-binding protein (follistatin) while germ (oogonia, oocytes) and somatic (theca, granulosa, luteal) cells express activin receptors, signaling components and inhibin co-receptor (betaglycan). Activins are implicated in various intra-ovarian roles including germ cell survival and primordial follicle assembly; follicle growth from preantral to mid-antral stages; suppression of thecal androgen production; promotion of granulosa cell proliferation, FSHR and CYP19A1 expression; enhancement of oocyte developmental competence; retardation of follicle luteinization and/or atresia and involvement in luteolysis. Inhibins (primarily inhibin A) are produced in greatest amounts by preovulatory follicles (and corpus luteum in primates) and suppress FSH secretion through endocrine negative feedback. Together with follistatin, inhibins act locally to oppose auto-/paracrine activin (and BMP) signaling thus modulating many of the above processes. The balance between activin-inhibin shifts during follicle development with activin signalling prevailing at earlier stages but declining as inhibin and betaglycan expression rise.


[Mesothelioma of Unknown Primary Site with Resection of Mediastinal Lymph Node Metastasis].
A 63-year-old male underwent biopsy of a mediastinal lymph node. Intra-operative frozen section diagnosis was metastatic mesothelioma. The primary site of mesothelioma was not detected in the thoracic cavity. One year later, he was again removed a mediastinal lymph node metastasis, but the primary site was not detected. Eight months later, he was performed right neck and mediastinal lymph node dissection due to additionally appeared lymph node metastases, but the primary site was not detected. He is well 6 months after last surgery with postoperative adjuvant chemotherapy.


Does cone-beam CT alter treatment plans? Comparison of preoperative implant planning using panoramic versus cone-beam CT images.
The present study was performed to compare the planning of implant placement based on panoramic radiography (PAN) and cone-beam computed tomography (CBCT) images, and to study the impact of the image dataset on the treatment planning. One hundred five partially edentulous patients (77 males, 28 females, mean age: 46 years, range: 26-67 years) seeking oral implant rehabilitation were referred for presurgical imaging. Imaging consisted of PAN and CBCT imaging. Four observers planned implant treatment based on the two-dimensional (2D) image datasets and at least one month later on the three-dimensional (3D) image dataset. Apart from presurgical diagnostic and dimensional measurement tasks, the observers needed to indicate the surgical confidence levels and assess the image quality in relation to the presurgical needs. All observers confirmed that both imaging modalities (PAN and CBCT) gave similar values when planning implant diameter. Also, the results showed no differences between both imaging modalities for the length of implants with an anterior location. However, significant differences were found in the length of implants with a posterior location. For implant dimensions, longer lengths of the implants were planned with PAN, as confirmed by two observers. CBCT provided images with improved scores for subjective image quality and surgical confidence levels. Within the limitations of this study, there was a trend toward PAN-based preoperative planning of implant placement leading towards the use of longer implants within the posterior jaw bone.


The interplay of cis-regulatory elements rules circadian rhythms in mouse liver.
The mammalian circadian clock is driven by cell-autonomous transcriptional feedback loops that involve E-boxes, D-boxes, and ROR-elements. In peripheral organs, circadian rhythms are additionally affected by systemic factors. We show that intrinsic combinatorial gene regulation governs the liver clock. With a temporal resolution of 2 h, we measured the expression of 21 clock genes in mouse liver under constant darkness and equinoctial light-dark cycles. Based on these data and known transcription factor binding sites, we develop a six-variable gene regulatory network. The transcriptional feedback loops are represented by equations with time-delayed variables, which substantially simplifies modelling of intermediate protein dynamics. Our model accurately reproduces measured phases, amplitudes, and waveforms of clock genes. Analysis of the network reveals properties of the clock: overcritical delays generate oscillations; synergy of inhibition and activation enhances amplitudes; and combinatorial modulation of transcription controls the phases. The agreement of measurements and simulations suggests that the intrinsic gene regulatory network primarily determines the circadian clock in liver, whereas systemic cues such as light-dark cycles serve to fine-tune the rhythms.


Comprehensive structural analysis of the open and closed conformations of Theileria annulata enolase by molecular modelling and docking.
Theileria annulata is an apicomplexan parasite which is responsible for tropical theileriosis in cattle. Due to resistance of T. annulata against commonly used antitheilerial drug, new drug candidates should be identified urgently. Enolase might be a druggable protein candidate which has an important role in glycolysis, and could also be related to several cellular functions as a moonlight protein. In this study; we have described three-dimensional models of open and closed conformations of T. annulata enolase by homology modeling method for the first time with the comprehensive domain, active site and docking analyses. Our results show that the enolase has similar folding patterns within enolase superfamily with conserved catalytic loops and active site residues. We have described specific insertions, possible plasminogen binding sites, electrostatic potential surfaces and positively charged pockets as druggable regions in T. annulata enolase.


Folding of an MHC class II-restricted tumor antigen controls its antigenicity via MHC-guided processing.
CD4(+) and CD8(+) T cell responses to endogenous retroviral envelope glycoprotein gp90 generate protective immunity to murine colon carcinoma CT26. A panel of I-A(d)-restricted T cell hybridomas recognize gp90 synthesized by CT26 cells but not by other gp90-expressing tumors. Here we report that antigenicity resides in an incompletely folded form of gp90 that is unique to CT26. In contrast to more compact forms of gp90 that are present in other tumors, this open conformer is captured by recycling I-A(d) on antigen-presenting cells and is processed intracellularly. Thus, gp90 acquires immunodominance via MHC-guided processing, and the generation of an MHC class II-restricted response can be controlled by the intracellular folding environment of antigen-expressing cells.


Identification and characterization of pbpC, the gene encoding Bacillus subtilis penicillin-binding protein 3.
Penicillin-binding proteins (PBPs) are enzymes involved in the synthesis of peptidoglycan structures in Bacillus subtilis such as the vegetative cell wall and the spore cortex. The B. subtilis sequencing project has identified a gene (orf16, EMBL accession number D38161) which exhibits significant sequence similarity to genes encoding class B high-molecular-weight PBPs. We have found that orf16 encodes PBP3 and have renamed this locus pbpC. Transcriptional fusions to lacZ were used to demonstrate that pbpC is transcribed primarily during log-phase growth, with lower amounts expressed during sporulation. During spore germination and outgrowth, pbpC expression resumes coincident with an increase in the optical density of the culture. The major promoter for pbpC is located just upstream of the gene; a low level of expression during sporulation appears to originate from much further upstream. Loss of PBP3 does not produce any detectable change in phenotype with respect to cell morphology, growth, sporulation, spore heat resistance, or spore germination and outgrowth. This was also true when the pbpC mutation was combined with mutations affecting other PBP-encoding genes to produce double mutants. These findings are consistent with previous evidence that many PBPs of B. subtilis have redundant functions within the cell.


Regulation of Monocyte Adhesion and Migration by Nox4.
We showed that metabolic disorders promote thiol oxidative stress in monocytes, priming monocytes for accelerated chemokine-induced recruitment, and accumulation at sites of vascular injury and the progression of atherosclerosis. The aim of this study was to identify both the source of reactive oxygen species (ROS) responsible for thiol oxidation in primed and dysfunctional monocytes and the molecular mechanisms through which ROS accelerate the migration and recruitment of monocyte-derived macrophages. We found that Nox4, a recently identified NADPH oxidase in monocytes and macrophages, localized to focal adhesions and the actin cytoskeleton, and associated with phospho-FAK, paxillin, and actin, implicating Nox4 in the regulation of monocyte adhesion and migration. We also identified Nox4 as a new, metabolic stress-inducible source of ROS that controls actin S-glutathionylation and turnover in monocytes and macrophages, providing a novel mechanistic link between Nox4-derived H2O2 and monocyte adhesion and migration. Actin associated with Nox4 was S-glutathionylated, and Nox4 association with actin was enhanced in metabolically-stressed monocytes. Metabolic stress induced Nox4 and accelerated monocyte adhesion and chemotaxis in a Nox4-dependent mechanism. In conclusion, our data suggest that monocytic Nox4 is a central regulator of actin dynamics, and induction of Nox4 is the rate-limiting step in metabolic stress-induced monocyte priming and dysfunction associated with accelerated atherosclerosis and the progression of atherosclerotic plaques.


Rat cytochromes P450 oxidize 2-nitrophenol, a human metabolite of carcinogenic 2-nitroanisole.
2-Nitrophenol (2-NP) is the major detoxification metabolite of an important industrial pollutant and a potent carcinogen, 2-nitroanisole (2-NA). Characterization of the products of 2-NP metabolism by rat hepatic microsomes containing cytochromes P450 (CYPs) and identification of the major CYP enzymes participating in this process are aims of this study. HPLC with UV detection was employed for the separation and characterization of 2-NP metabolites. Inducers and inhibitors of CYPs and rat recombinant CYPs were used to characterize the enzymes participating in 2-NP oxidation. Rat hepatic microsomes oxidize 2-NP to its hydroxylated metabolite, 2,5-dihydroxynitrobenzene (2,5-DNB). No nitroreductive metabolism leading to the formation of o-aminophenol was evident when using rat hepatic microsomes. Selective CYP inhibitors and hepatic microsomes of rats pre-treated with specific CYP inducers were used to characterize CYPs oxidizing 2-NP in rat livers. Based on these studies, we attribute most of 2-NP oxidation in rat liver to CYP2E1 and 3A, followed by CYP2D and 2C. Among recombinant rat CYP enzymes tested in this study, CYP2E1 and 2C11 were the most effective enzymes oxidizing 2-NP. Oxidation of 2-NP by rat CYP2E1 exhibits the Michaelis-Menten kinetics, having the Km value of 0.35 mM. The results found in this study, the first report on the metabolism of 2-NP by rat hepatic microsomes and rat CYP enzymes, demonstrate that CYP2E1 is the major enzyme oxidizing this compound in rat liver.


French vertical-flow constructed wetland design: adaptations for tropical climates.
The French Outermost Regions are under tropical climate yet still have to comply with both French and EU regulations. French vertical-flow constructed wetland systems appear well adapted to the technical specifics of these regions but their adaptation to tropical climate requires new design guidelines to be defined (area needed, number of filters, type of plants, material to be used, etc.). A study was started in 2008, with backing from the national water authorities, to implement full-scale experimental sites and assess the impacts of local context on design and performances. This paper reports the monitoring results on three vertical-flow constructed wetlands fed directly with raw wastewater (known as the 'French system') in Mayotte and French Guiana. The plants, now in operation for between 1 and 6 years, range from 160 to 480 population equivalent (p.e.). Monitoring consisted of 28 daily composite flow samples in different seasons (dry season, rainy season) at the inlet and outlet of each filter. Performances are benchmarked against French mainland area standards from Irstea's database. Results show that performances are improved by warmer temperature for chemical oxygen demand (COD), suspended solids (SS) and total Kjeldahl nitrogen (TKN) and satisfy national quality objectives with a single stage of filters. Treatment plant footprint can thus be reduced as only two parallel filters are needed. Indeed, warm temperatures allow faster mineralization of the sludge deposit, making it possible to operate at similar rest and feeding period durations. Systems operated using one twin-filter stage can achieve over 90% COD, SS and TKN removal for a total surface of 0.8 m²/p.e.


Genetic interactions between Shox2 and Hox genes during the regional growth and development of the mouse limb.
The growth and development of the vertebrate limb relies on homeobox genes of the Hox and Shox families, with their independent mutation often giving dose-dependent effects. Here we investigate whether Shox2 and Hox genes function together during mouse limb development by modulating their relative dosage and examining the limb for nonadditive effects on growth. Using double mRNA fluorescence in situ hybridization (FISH) in single embryos, we first show that Shox2 and Hox genes have associated spatial expression dynamics, with Shox2 expression restricted to the proximal limb along with Hoxd9 and Hoxa11 expression, juxtaposing the distal expression of Hoxa13 and Hoxd13. By generating mice with all possible dosage combinations of mutant Shox2 alleles and HoxA/D cluster deletions, we then show that their coordinated proximal limb expression is critical to generate normally proportioned limb segments. These epistatic interactions tune limb length, where Shox2 underexpression enhances, and Shox2 overexpression suppresses, Hox-mutant phenotypes. Disruption of either Shox2 or Hox genes leads to a similar reduction in Runx2 expression in the developing humerus, suggesting their concerted action drives cartilage maturation during normal development. While we furthermore provide evidence that Hox gene function influences Shox2 expression, this regulation is limited in extent and is unlikely on its own to be a major explanation for their genetic interaction. Given the similar effect of human SHOX mutations on regional limb growth, Shox and Hox genes may generally function as genetic interaction partners during the growth and development of the proximal vertebrate limb.


Dinucleotide-sensing proteins: linking signaling networks and regulating transcription.
Differential binding of dinucleotides to key regulatory proteins can modulate their interactions with other proteins and, in some cases, can signal fluctuations in the cellular redox state, to produce changes in transcription and physiological state. The dinucleotide-binding proteins human HSCARG and yeast transcription repressor Gal80p are examples that offer exciting glimpses into the potential for dinucleotide-sensing proteins to couple fluctuations in dinucleotide ratios to changes in transcription and to act as networking agents linking different classes of signaling molecules.


Nest predation increases with parental activity: separating nest site and parental activity effects.
Alexander Skutch hypothesized that increased parental activity can increase the risk of nest predation. We tested this hypothesis using ten open-nesting bird species in Arizona, USA. Parental activity was greater during the nestling than incubation stage because parents visited the nest frequently to feed their young during the nestling stage. However, nest predation did not generally increase with parental activity between nesting stages across the ten study species. Previous investigators have found similar results. We tested whether nest site effects might yield higher predation during incubation because the most obvious sites are depredated most rapidly. We conducted experiments using nest sites from the previous year to remove parental activity. Our results showed that nest sites have highly repeatable effects on nest predation risk; poor nest sites incurred rapid predation and caused predation rates to be greater during the incubation than nestling stage. This pattern also was exhibited in a bird species with similar (i.e. controlled) parental activity between nesting stages. Once nest site effects are taken into account, nest predation shows a strong proximate increase with parental activity during the nestling stage within and across species. Parental activity and nest sites exert antagonistic influences on current estimates of nest predation between nesting stages and both must be considered in order to understand current patterns of nest predation, which is an important source of natural selection.


Novel photocleavable universal support for oligonucleotide synthesis.
A novel photocleavable universal support for the automated solid phase synthesis of oligonucleotides is described. The linker between the growing oligonucleotide chain and CPG support contains a nucleophilic amine protected with a photocleavable group. On exposure to UV light, this group is detached and the free amine affords cleavage of the oligonucleotide from the support. The use of long wavelength UV light avoids damage to the DNA.