Evaluation of vegetation size and its relationship with septic pulmonary embolism in tricuspid valve infective endocarditis: A real time 3DTEE study.
Tricuspid valve infective endocarditis (TVIE) causes septic pulmonary embolism (PE). However, the impact of vegetation size on PE is not fully elucidated. In 26 consecutive patients with definite TVIE according to Duke criteria, we tested clinical, microbiological, and echocardiographic data including real time three-dimensional transesophageal echocardiography (3DTEE) as potential predictors of PE. 3DTEE measurement of maximum length of vegetation (MLV) was obtained with Advanced QLAB Quantification Software by cropping the 3D volume with the appropriate 2D plane to obtain the largest value. The standard two-dimensional transesophageal echocardiography (2DTEE) images were also evaluated to determine the MLV. Pulmonary embolism occurred after TVIE diagnosis with 3DTEE assessment and initiation of antibiotic therapy in 12 patients (46.2%). The 3DTEE MLV was larger than the 2DTEE value with a mean difference of 3.6 mm (95% CI, 2.5-4.6 mm). The best cutoff value for prediction of PE was MLV ≥16.4 mm with 3DTEE and MLV ≥9.5 mm with 2DTEE. The positive predictive value increased from 76.9% to 90% when 3DTEE was used. The accuracy of classification of patients with PE increased from 80.6% to 84.9% with 3DTEE. On multivariate analysis, 3DTEE MLV ≥16.4 mm (odds ratio 20.5; 95% confidence interval 1.31-322; P=.031) was independently associated with the occurrence of PE after adjustment for age, sex, complex vegetation, and the number of vegetation-attached leaflets. In TVIE, vegetation length is a strong predictor of the occurrence of PE. In combination with 2DTEE, 3DTEE may identify high-risk patients who will need a more aggressive therapeutic strategy.


MAD phasing using the (Ta6Br12)2+ cluster: a retrospective study.
The crystal structure of cytokinin-specific binding protein (CSBP) containing four independent molecules with 4 x 155 = 620 residues in the asymmetric unit of the P6(4) unit cell has been solved by three-wavelength MAD using 1.8 angstroms resolution data recorded from a crystal derivatized with the dodecabromohexatantalum cation (Ta6Br12)2+. The diffraction data contained a very strong anomalous signal (allowing successful phasing even using peak SAD data alone) despite the fact that the five (Ta6Br12)2+ clusters found in the asymmetric unit have low occupancy (about 0.3). The derivative structure has been successfully refined to R = 0.158, providing interesting details on the geometry of the (Ta6Br12)2+ cluster, its interactions with the protein and on the backsoaking of a cytokinin ligand that was originally part of a CSBP-cytokinin complex in the native crystals used for (Ta6Br12)2+ derivatization. A simulation analysis of the phasing power of the (Ta6Br12)2+ ions at artificially imposed resolution limits shows that it is not possible to resolve the individual Ta atoms if the dmin limit of the data is higher than 2.9 angstroms. Additionally, for successful Ta identification the (Ta6Br12)2+ complex should be specifically bound and ordered. Good binding at the protein surface is facilitated by the presence of acidic groups, indicating higher pH buffer conditions to be preferable. In addition, the water channels in the crystal should be sufficiently wide (at least 11 angstroms) to allow free diffusion of the (Ta6Br12)2+ ions on soaking. A retrospective look at the initial molecular-replacement calculations provides interesting insights into how the peculiar packing mode and strong bias of the molecular-replacement-phased electron-density maps had hindered successful solution of the structure by this method.


Social context and psychosocial influences on blood pressure among American Samoans.
This study explores social and explores social and economic influences on health within a model formulated to address explicitly both individual and household level phenomena. Dressler's lifestyle incongruity model is used as a basis from which to predict the effects of intracultural contexts of variability on blood pressure. The sample for this survey consists of 134 Samoan men and women living in American Samoa. Based on previous experience and ethnographic sources, two key intracultural contexts were examined; gender, i.e., male-female differences in response to psychosocial stress, and household employment as indicated by whether or not both spouses in a household are employed. Our analysis indicates that lifestyle incongruity, defined as the difference between the material culture presented by a household and the economic resources of the family, is significantly associated with both systolic and diastolic blood pressure. Furthermore, males and females show opposite blood pressure associations with both lifestyle incongruity (male blood pressure increases with increasing incongruity while female blood pressure does not) and household employment (male blood pressure is higher when both spouses work but female blood pressure is lower).


Effects of a sulfonylurea herbicide on the soil bacterial community.
Sulfonylurea herbicides are widely used on a wide range of crops to control weeds. Chevalier® OnePass herbicide is a sulfonylurea herbicide intensively used on cereal crops in Algeria. No information is yet available about the biodegradation of this herbicide or about its effect on the bacterial community of the soil. In this study, we collected an untreated soil sample, and another sample was collected 1 month after treatment with the herbicide. Using a high-resolution melting DNA technique, we have shown that treatment with Chevalier® OnePass herbicide only slightly changed the composition of the whole bacterial community. Two hundred fifty-nine macroscopically different clones were isolated from the untreated and treated soil under both aerobic and microaerobic conditions. The strains were identified by sequencing a conserved fragment of the 16S rRNA gene. The phylogenetic trees constructed using the sequencing results confirmed that the bacterial populations were similar in the two soil samples. Species belonging to the Lysinibacillus, Bacillus, Pseudomonas, and Paenibacillus genera were the most abundant species found. Surprisingly, we found that among ten strains isolated from the treated soil, only six were resistant to the herbicide. Furthermore, bacterial overlay experiments showed that only one resistant strain (related to Stenotrophomonas maltophilia) allowed all the sensitive strains tested to grow in the presence of the herbicide. The other resistant strains allowed only certain sensitive strains to grow. On the basis of these results, we propose that there must be several biodegradation pathways for this sulfonylurea herbicide.


Behavioural responses of fin whales to military mid-frequency active sonar.
The effect of active sonars on marine mammal behaviour is a topic of considerable interest and scientific investigation. Some whales, including the largest species (blue whales, Balaenoptera musculus), can be impacted by mid-frequency (1-10 kHz) military sonars. Here we apply complementary experimental methods to provide the first experimentally controlled measurements of behavioural responses to military sonar and similar stimuli for a related endangered species, fin whales (Balaenoptera physalus). Analytical methods include: (i) principal component analysis paired with generalized additive mixed models; (ii) hidden Markov models; and (iii) structured expert elicitation using response severity metrics. These approaches provide complementary perspectives on the nature of potential changes within and across individuals. Behavioural changes were detected in five of 15 whales during controlled exposure experiments using mid-frequency active sonar or pseudorandom noise of similar frequency, duration and source and received level. No changes were detected during six control (no noise) sequences. Overall responses were more limited in occurrence, severity and duration than in blue whales and were less dependent upon contextual aspects of exposure and more contingent upon exposure received level. Quantifying the factors influencing marine mammal responses to sonar is critical in assessing and mitigating future impacts.


Predictive Modeling of a Batch Filter Mating Process.
Quantitative characterizations of horizontal gene transfer are needed to accurately describe gene transfer processes in natural and engineered systems. A number of approaches to the quantitative description of plasmid conjugation have appeared in the literature. In this study, we seek to extend that work, motivated by the question of whether a mathematical model can accurately predict growth and conjugation dynamics in a batch process. We used flow cytometry to make time-point observations of a filter-associated mating between two E. coli strains, and fit ordinary differential equation models to the data. A model comparison analysis identified the model formulation that is best supported by the data. Identifiability analysis revealed that the parameters were estimated with acceptable accuracy. The predictive power of the model was assessed by comparison with test data that demanded extrapolation from the training experiments. This study represents the first attempt to assess the quality of model predictions for plasmid conjugation. Our successful application of this approach lays a foundation for predictive modeling that can be used both in the study of natural plasmid transmission and in model-based design of engineering approaches that employ conjugation, such as plasmid-mediated bioaugmentation.


Evaluation of Pillars4life: a virtual coping skills program for cancer survivors.
Pillars4Life is an educational program that teaches coping skills to cancer patients in a virtual group setting; it was recently implemented at 17 hospitals across the USA. The cost-effective, scalable, and assessable Pillars4Life curriculum targets psychosocial resources (e.g., self-efficacy and coping skills) as a means to reduce symptoms (e.g., depression, anxiety, and posttraumatic stress) and enhance quality of life. Cancer patients were recruited from hospitals that received the LIVESTRONG Community Impact Project Award to enroll in a pilot study of Pillars4Life. Consenting participants met with a certified instructor weekly for 10 weeks in a virtual environment; the manualized intervention trained participants in personal coping skills. Longitudinal assessments over 6 months were assessed using validated instruments to determine changes in Pillars4Life targeted resources and outcomes. Multiple linear regression models examined the relationship between changes in targeted resources and changes in outcome from baseline to 3 months post-intervention. Participants (n = 130) had the following characteristics: mean age of 56 ± 11 years, 87% women, 11% non-Caucasian, and 77% with college degree. At 3- and 6-month follow-up, mean scores improved on all key outcome measures such as depression (Patient Health Questionnaire), anxiety (Generalized Anxiety Disorder), posttraumatic stress (Posttraumatic Stress Disorder Checklist), fatigue (Functional Assessment of Chronic Illness Therapy-Fatigue), and well-being (Functional Assessment of Cancer Therapy-General) from baseline (all p < 0.01); results were most pronounced among participants who reported ≥4/10 on the Distress Thermometer at baseline (all p < 0.001). Changes in each targeted resource were associated with 3-month improvements in at least one outcome. Participation in the Pillars4Life program was associated with statistically and clinically significant improvements in scores on pre-specified outcomes and targeted resources.


Determination of 18 phenolic acids in tobacco and rhizosphere soil by ultra high performance liquid chromatography combined with triple quadrupole mass spectrometry.
An ultra high performance liquid chromatography with triple quadrupole mass spectrometry method for the determination of free and bound phenolic acids in tobacco plant and soil was developed. A simple solid-phase extraction, which used Polar Enhanced Polymer column as stationary phase and methanol as mobile phase, was used for the clean-up of bound phenolic acids, and a liquid-phase extraction using chloroform as solvent was used to purify free phenolic acids. With our method, 18 phenolic acids in rhizosphere soil of continuous cropping flue-cured cultivar k326 were separated and determined within 6 min with recoveries of 82-107% and relative standard deviations (n = 5) of 1.1-4.8%. Results showed that free phenolic acids accounted for 0-9, 92-100, and 69-100% of total phenolic acids in rhizosphere soil, cultivar k326 roots and leaves, respectively. Results also revealed that p-hydroxybenzoic acid, p-coumaric acid, vanillic acid, ferulic acid, and syringic acid were the predominant phenolic acids in rhizosphere soil of cultivar k326, and continuous cropping of cultivar k326 in the same farmland could lead to the accumulation of these phenolic acids in soil except syringic acid. The determination of phenolic acids provided detailed information for evaluating their source and characteristics in continuous cropping tobacco plant and soil.


Are there Field Triage Criteria that Can Predict Low-Yield Air Medical Transports?
Air medical transport of trauma patients from the scene of injury plays a critical role in the delivery of severely injured patients to trauma centers. Over-triage of patients to trauma centers reduces the system efficiency and jeopardizes safety of air medical crews. The objective of this study was to determine which triage factors utilized by Emergency Medical Services (EMS) providers are strong predictors of early discharge for trauma patients transported by helicopter to a trauma center. A retrospective chart review over a two-year period was performed for trauma patients flown from the injury site into a Level I trauma center by an air medical transport program. Demographic and clinical data were collected on each patient. Prehospital factors such as Glasgow Coma Score (GCS), Revised Trauma Score (RTS), intubation status, mechanism of injury, anatomic injuries, physiologic parameters, and any combinations of these factors were investigated to determine which triage criteria accurately predicted early discharge. Hospital factors such as Injury Severity Score (ISS), length-of-stay (LOS), survival, and emergency department disposition were also collected. Early discharge was defined as a hospital stay of less than 24 hours in a patient who survives their injuries. A more stringent definition of appropriate triage was defined as a patient with in-hospital death, an ISS >15, those taken to the operating room (OR) or intensive care unit (ICU), or those receiving blood products. Those patients who failed to meet these criteria were also used to determine over-triage rates. An overall early discharge rate of 35% was found among the study population. Furthermore, when the more stringent definition was applied, over-triage rates were as high as 85%. Positive predictive values indicated that patients who met at least one anatomic and physiologic criteria were appropriately transported by helicopter as 94% of these patients had stays longer than 24 hours. No other criteria or combination of criteria had a high predictive value for early discharge. No individual triage criteria or combination of criteria examined demonstrated the ability to uniformly predict an early discharge. Although helicopter transport and subsequent hospital care is costly and resource consuming, it appears that a significant number of patients will be discharged within 24 hours of their transport to a trauma center. Future studies must determine the impact of eliminating "low-yield" triage criteria on under-triage of scene trauma patients.


Prior cytomegalovirus infection does not predict clinical outcome after percutaneous transluminal coronary angioplasty.
A direct association between human cytomegalovirus (HCMV) infection and the development of restenosis after coronary angioplasty has been suggested. The aim of this prospective study was to evaluate the value of HCMV serology in predicting the clinical outcome after percutaneous transluminal coronary angioplasty (PTCA). 112 patients undergoing elective PTCA were included in the study. HCMV antibody levels were measured by ELISA. Cardiac events within a follow-up period of 6 months after PTCA were defined as (1) progression or recurrence of anginal complaints and/or a positive exercise test; (2) restenosis that required repeat revascularization. 73% of PTCA patients were seropositive for HCMV. Successful PTCA was achieved in a total of 94 patients, who were followed for 6 months. In 31/94 patients (33%) cardiac events occurred and in 15/94 (16%), this could be related to restenosis. We found no statistically significant difference between seropositive and negative patients with respect to anginal complaints or the need for revascularization. There was no evidence of acute reactivation, since titers of anti-HCMV antibodies did not increase after PTCA. This study shows that the clinical outcome after PTCA is not related to the HCMV serostatus of the patient. Therefore, our data do not support the hypothesis that serological markers of HCMV infection are of clinical importance for the assessment of a patient's individual risk after PTCA. This does not preclude a role for local reactivation of HCMV at the site of angioplasty.


Where is an experienced medical educator when you need one?
This article was migrated. The article was marked as recommended.


Questions and replies: renal mechanisms for urinary concentrating and diluting processes.
Mechanisms for urinary concentration or dilution depend on counterflow processes, both tubular and vascular, within the renal medulla. Recently, there have emerged differing hypotheses about the renal tubular processes responsible for maintaining a hypertonic medullary interstitium. In this Editorial Review, R.W. Berliner frames three questions germane to this issue, and J.P. Kokko and D.J. Marsh provide their responses to these queries. The major issues addressed are: 1) What are the major unresolved question(s) concerning the mechanism by which concentrated urine is formed? 2) Current evidence suggests that the urea concentration in thin ascending limbs is slightly lower in the lumen than in interstitial fluid. Is the transepithelial concentration gradient between thin ascending limb and renal medullary interstitium sufficient to permit an entirely passive mechanism for diluting tubular fluid in the thin ascending limb? 3) A simple three-compartment model for the renal medullary concentrating process would include the tubular lumen, peritubular capillary, and the interstitium. Is it possible to generate a model that, by juxtaposing medullary structures, might explain renal medullary counterflow processes more adequately than the simple three-compartment model?


Human RecQ Helicases in DNA Double-Strand Break Repair.
RecQ DNA helicases are a conserved protein family found in bacteria, fungus, plants, and animals. These helicases play important roles in multiple cellular functions, including DNA replication, transcription, DNA repair, and telomere maintenance. Humans have five RecQ helicases: RECQL1, Bloom syndrome protein (BLM), Werner syndrome helicase (WRN), RECQL4, and RECQL5. Defects in BLM and WRN cause autosomal disorders: Bloom syndrome (BS) and Werner syndrome (WS), respectively. Mutations in RECQL4 are associated with three genetic disorders, Rothmund-Thomson syndrome (RTS), Baller-Gerold syndrome (BGS), and RAPADILINO syndrome. Although no genetic disorders have been reported due to loss of RECQL1 or RECQL5, dysfunction of either gene is associated with tumorigenesis. Multiple genetically independent pathways have evolved that mediate the repair of DNA double-strand break (DSB), and RecQ helicases play pivotal roles in each of them. The importance of DSB repair is supported by the observations that defective DSB repair can cause chromosomal aberrations, genomic instability, senescence, or cell death, which ultimately can lead to premature aging, neurodegeneration, or tumorigenesis. In this review, we will introduce the human RecQ helicase family, describe in detail their roles in DSB repair, and provide relevance between the dysfunction of RecQ helicases and human diseases.


[Reactivity of GST-SAPA antigen of Trypanosoma cruzi against sera from patients with Chagas disease and leishmaniasis].
Serologic diagnosis of Trypanosoma cruzi infection is important due to the limited sensitivity of direct parasitologic methods for diagnosis in the indeterminate and chronic phases of disease. SAPA antigen has been used in several studies and has been shown to be a good marker for use in the diagnosis of T. cruzi infection. Chagas disease and leishmaniasis are endemic in northern Salta with overlapping zones of transmission, which frequently leads to T. cruzi-Leishmania spp. mixed infections. Diagnosis is complicated by the fact that there is significant cross-reactivity when non-specific antigens are used. We evaluated the reactivity of GST-SAPA antigen in the ELISA test (ELISA-SAPA) against sera from persons infected with T. cruzi (n = 154), leishmaniasis (n = 66), mixed infections (29), and healthy controls (n = 28) using commercial ELISA and IHA kits as reference tests. For ELISA-SAPA the sensitivity, specificity and kappa index were calculated for detection of T. cruzi infection. Among sera from patients infected with leishmaniasis, 30.5% of co-infections were detected. ELISA-SAPA sensitivity was 97.1% (confidence interval 95%: 94.5-99.9), specificity was 100% (confidence interval 95%: 99.4-100), and kappa index was 96% (confidence interval 95%: 93-99%), for detection of T. cruzi infection. Sensitivity, specificity and kappa indices have shown a high efficiency of ELISA-SAPA.


Cytoprotective effects of xanthohumol against methylglyoxal-induced cytotoxicity in MC3T3-E1 osteoblastic cells.
Methylglyoxal (MG) has been suggested to be a major source of intracellular reactive carbonyl compounds, and has been implicated in increasing the levels of advanced glycation end products in age-related diseases. Xanthohumol is a prenylated flavonoid found in hops (Humulus lupulus) and beer. In the present study, we investigated the effects of xanthohumol on MG-induced cytotoxicity in osteoblastic MC3T3-E1 cells. Xanthohumol attenuated MG-induced cytotoxicity, as evidenced by improved cell viability, and prevented MG-induced MG-protein adducts, inflammatory cytokines, reactive oxygen species and mitochondrial superoxide production. In addition, xanthohumol increased glyoxalase I activity, glutathione, heme oxygenase-1 and nuclear factor erythroid 2-related factor 2 levels in the presence of MG. Pretreatment with xanthohumol before MG exposure reduced MG-induced mitochondrial dysfunction. Furthermore, xanthohumol treatment resulted in a significant reduction in the levels of endoplasmic reticulum stress and autophagy induced by MG. Notably, the autophagy-reducing effect of xanthohumol was abolished after the addition of Ex527, a selective inhibitor of sirtuin 1, suggesting that xanthohumol is an effective sirtuin 1 activator for reducing autophagy. Taken together, our findings suggest xanthohumol as a promising new strategy for preventing diabetic osteopathy.


Language style matching and treatment outcome in anorexia nervosa.
Psychotherapy is the treatment of choice for anorexia nervosa (AN) but mechanisms of action are still largely unknown. Growing research suggests that synchrony between patient and therapist contributes to treatment success. Adding to this literature, this study examined the association between language style matching (LSM) among patient and therapist as an indicator of interpersonal (verbal) synchrony and treatment outcome in AN. Audio recordings of mid-treatment therapy sessions (n = 25) in a multi-centre randomized controlled trial on the Maudsley Model of Anorexia Nervosa Treatment for Adults were transcribed and used to calculate LSM for each patient-therapist dyad. These scores were used to predict treatment outcome at 12-month follow-up. LSM did not predict body mass index (primary outcome) at follow-up. However, higher LSM (M = 0.87, SD = 0.04) was associated with lower eating disorder psychopathology (accounting for 11% of the variance) and higher recovery rates (accounting for 28% of variance) at follow-up. These preliminary findings suggest that verbal synchrony between patients with AN and their therapists contributes to favourable treatment outcomes. High levels of LSM may reflect therapeutic empathy, cooperation, or mutual positive perception. Further research should explore the mechanisms of linguistic synchrony with larger samples to allow for stronger conclusions.


Characterization of Staphylococcus aureus strains associated with food poisoning outbreaks in France.
Enterotoxins produced by Staphylococcus aureus are responsible for staphylococcal food-poisoning outbreaks (SFPO). In France, SFPO are the second cause of food-borne diseases after Salmonella. However, very little is known about the strains involved. The objective of this study was to characterize the staphylococcal strains related to these SFPO through phenotypic and genotypic analyses. A total of 178 coagulase-positive staphylococcal isolates recovered from 31 SFPO (1981-2002) were screened through biotyping. Thirty-three strains representative of the different biotypes in each SFPO were further examined for SmaI macrorestriction-type, phage-type, resistance to various antimicrobial drugs, presence of staphylococcal enterotoxin (se) genes sea to sei, and production of enterotoxins SEA to SED. All these 33 strains were identified as S. aureus species: 27 were of human biotypes and six ovine or non-host-specific biotypes. Most (74.1%) strains reacted with group III phages. Eleven strains were resistant to at least two classes of antibiotics and among them, two were resistant to methicillin. Twenty-nine strains carried one or several of the eight se genes tested; the gene sea was most common (n=23), and often linked to sed (n=12) or seh (n=5). The novel se genes seg-i were in all cases associated with se genes sea to sed except for one strain which carried only seg and sei. Pulsed-Field Gel Electrophoresis (PFGE) of SmaI macrorestriction digests of the 33 strains discriminated 32 PFGE patterns grouped into nine biotype-specific clusters. All five strains carrying sea and seh were grouped together into the same sub-cluster. Three of the four se-gene-negative strains were in one PFGE cluster: all four should be tested for se genes not included in this study and, if negative, be further investigated for the presence of unidentified SEs.


Preconception counseling and contraception after gestational diabetes.
Women with gestational diabetes mellitus (GDM) diagnosed in the period 1978-1984 were followed for on average 6 yr after the index pregnancy. Thirty percent had diabetes mellitus at the follow-up examination, and preliminary results indicate that at least another third will develop diabetes during a subsequent pregnancy. Therefore, family planning and contraceptive guidance should follow the lines for women with pregestational diabetes. When low-dose hormonal contraceptives containing ethinyl estradiol and levonorgestrel were given to women with previous GDM, glucose tolerance and lipoprotein levels remained unchanged during a 6-mo treatment. However, insulin response to oral glucose increased significantly after hormonal intake for 6 mo. A triphasic preparation resulted in a significantly lower insulin response than a low-dose monophasic preparation. However, the results indicate that low-dose oral-contraceptive compounds appear to be safe for women with previous GDM when administered for limited periods. At the follow-up examination, we found no increased risk of developing diabetes in women with previous GDM who used oral contraception. We consider the intrauterine contraceptives (IUD) a safe and effective alternative for women with previous GDM. Of 154 women with GDM, 33% chose IUD, 22% a combination-type oral contraceptive, and 16% barrier methods as their first choice of contraception 2 mo postpartum. We conclude that family planning and qualified contraceptive advice are important in women with previous GDM.


Dyslexia and Developmental Language Disorder: comorbid disorders with distinct effects on reading comprehension.
Reading comprehension draws on both decoding and linguistic comprehension, and poor reading comprehension can be the consequence of a deficit in either of these skills. Using outcome data from the longitudinal Wellcome Language and Reading Project, we identified three groups of children at age 8 years: children with dyslexia (N = 21) who had deficits in decoding but not oral language, children with Developmental Language Disorder (DLD; N = 38) whose decoding skills were in the normal range, and children who met criteria for both dyslexia and DLD (N = 29). All three groups had reading comprehension difficulties at the ages of 8 and 9 years relative to TD controls though those of the children with dyslexia were mild (relative to TD controls, d = 0.51 at age 8, d = 0.60 at age 8); while the most severe problems were found in the comorbid dyslexia + DLD group (d = 1.79 at age 8, d = 2.06 at age 9) those with DLD also had significant difficulties (d = 1.56 at age 8, d = 1.56 at age 9). These findings confirm that children with dyslexia or DLD are at-risk for reading comprehension difficulties but for different reasons, because of weak decoding in the case of dyslexia or weak oral language skills in the case of DLD. Different forms of intervention are required for these groups of children, targeted to their particular area(s) of weakness.


Reconstructing ancestral gene content by coevolution.
Inferring the gene content of ancestral genomes is a fundamental challenge in molecular evolution. Due to the statistical nature of this problem, ancestral genomes inferred by the maximum likelihood (ML) or the maximum-parsimony (MP) methods are prone to considerable error rates. In general, these errors are difficult to abolish by using longer genomic sequences or by analyzing more taxa. This study describes a new approach for improving ancestral genome reconstruction, the ancestral coevolver (ACE), which utilizes coevolutionary information to improve the accuracy of such reconstructions over previous approaches. The principal idea is to reduce the potentially large solution space by choosing a single optimal (or near optimal) solution that is in accord with the coevolutionary relationships between protein families. Simulation experiments, both on artificial and real biological data, show that ACE yields a marked decrease in error rate compared with ML or MP. Applied to a large data set (95 organisms, 4873 protein families, and 10,000 coevolutionary relationships), some of the ancestral genomes reconstructed by ACE were remarkably different in their gene content from those reconstructed by ML or MP alone (more than 10% in some nodes). These reconstructions, while having almost similar likelihood/parsimony scores as those obtained with ML/MP, had markedly higher concordance with the coevolutionary information. Specifically, when ACE was implemented to improve the results of ML, it added a large number of proteins to those encoded by LUCA (last universal common ancestor), most of them ribosomal proteins and components of the F(0)F(1)-type ATP synthase/ATPases, complexes that are vital in most living organisms. Our analysis suggests that LUCA appears to have been bacterial-like and had a genome size similar to the genome sizes of many extant organisms.


Assessing the Dim Light Melatonin Onset in Adults with Autism Spectrum Disorder and No Comorbid Intellectual Disability.
This study assessed melatonin levels and the dim light melatonin onset (DLMO) in adults with Autism Spectrum Disorder (ASD) and also investigated the relationships between melatonin and objectively measured sleep parameters. Sixteen adults with ASD (ASD-Only), 12 adults with ASD medicated for comorbid diagnoses of anxiety and/or depression (ASD-Med) and 32 controls participated in the study. Although, the timing of the DLMO did not differ between the two groups, advances and delays of the melatonin rhythm were observed in individual profiles. Overall mean melatonin levels were lower in the ASD-Med group compared to the two other groups. Lastly, greater increases in melatonin in the hour prior to sleep were associated with greater sleep efficiency in the ASD groups.


BMP antagonism is required in both the node and lateral plate mesoderm for mammalian left-right axis establishment.
In mouse, left-right (L-R) patterning depends on asymmetric expression of Nodal around the node, leading to Nodal expression specifically in the left lateral plate mesoderm (LPM). Bone morphogenetic protein (BMP) signaling is also involved, but the mechanistic relationship with Nodal expression remains unclear. We find that BMP signal transduction is higher in the right LPM, although Bmp4, which is required for L-R patterning, is expressed symmetrically. By contrast, the BMP antagonists noggin (Nog) and chordin (Chrd) are expressed at higher levels in the left LPM. In Chrd;Nog double mutants, BMP signaling is elevated on both sides, whereas Nodal expression is absent. Ectopic expression of Nog in the left LPM of double mutants restores Nodal expression. Ectopic Bmp4 expression in the left LPM of wild-type embryos represses Nodal transcription, whereas ectopic Nog in the right LPM leads to inappropriate Nodal expression. These data indicate that chordin and noggin function to limit BMP signaling in the left LPM, thereby derepressing Nodal expression. In the node, they promote peripheral Nodal expression and proper node morphology, potentially in concert with Notch signaling. These results indicate that BMP antagonism is required in both the node and LPM to facilitate L-R axis establishment in the mammalian embryo.


Ten Environmental Sustainability Initiatives for Implementation in Human-Service Organizations.
Climate change is happening at an escalating rate. Every organization, including human-service organizations, needs to determine how their policies and practices positively and negatively affect the environment and enact a plan for reducing their negative impacts. There is no targeted guidance for leaders of human-service organizations, like applied behavior analysis organizations, to reduce their negative impact on the environment. This article provides 10 initiatives human service organizations can implement toward this goal.


Dual Antiplatelet Therapy Duration Determines Outcome After 2- But Not 1-Stent Strategy in Left Main Bifurcation Percutaneous Coronary Intervention.
The aim of this study was to investigate clinical outcomes after left main coronary artery (LM) bifurcation percutaneous coronary intervention (PCI) and the impact of the duration of dual antiplatelet therapy (DAPT) according to treatment strategy. There are limited data regarding the optimal PCI strategy for LM bifurcation lesions with new-generation drug-eluting stents. A patient-level pooled analysis of 5 nationwide multicenter registries was performed. Rates of target lesion failure, thrombotic adverse cardiovascular events, and their individual components at 3-year were analyzed. Subgroup analysis according to DAPT duration was performed. From 13,172 patients undergoing PCI with new-generation drug-eluting stents, a total of 700 patients were treated for LM bifurcation lesions, 567 with a 1-stent strategy and 133 with a 2-stent strategy. Rates of target lesion failure and target lesion revascularization were higher in the 2-stent group, driven mainly by complex lesion profiles. Risks for thrombotic adverse cardiovascular events and its components were comparable between the 2 strategies. Subgroup analysis showed that risks for target lesion failure and thrombotic adverse cardiovascular events in the 2-stent group were significantly higher than in the 1-stent group in those with DAPT interruption <1 year, while they were similar in those receiving DAPT maintenance ≥1 year. Up to 20% of patients who underwent LM bifurcation PCI eventually required a 2-stent strategy, which was as safe as a 1-stent strategy with the use of new-generation drug-eluting stents. Careful pre-emptive case selection as well as prolonged DAPT may be necessary when considering a 2-stent strategy in LM PCI given its higher rate of repeat revascularization and lesion failure than the 1-stent approach.


Mapping longitudinal scientific progress, collaboration and impact of the Alzheimer's disease neuroimaging initiative.
Alzheimer's disease neuroimaging initiative (ADNI) is a landmark imaging and omics study in AD. ADNI research literature has increased substantially over the past decade, which poses challenges for effectively communicating information about the results and impact of ADNI-related studies. In this work, we employed advanced information visualization techniques to perform a comprehensive and systematic mapping of the ADNI scientific growth and impact over a period of 12 years. Citation information of ADNI-related publications from 01/01/2003 to 05/12/2015 were downloaded from the Scopus database. Five fields, including authors, years, affiliations, sources (journals), and keywords, were extracted and preprocessed. Statistical analyses were performed on basic publication data as well as journal and citations information. Science mapping workflows were conducted using the Science of Science (Sci2) Tool to generate geospatial, topical, and collaboration visualizations at the micro (individual) to macro (global) levels such as geospatial layouts of institutional collaboration networks, keyword co-occurrence networks, and author collaboration networks evolving over time. During the studied period, 996 ADNI manuscripts were published across 233 journals and conference proceedings. The number of publications grew linearly from 2008 to 2015, so did the number of involved institutions. ADNI publications received much more citations than typical papers from the same set of journals. Collaborations were visualized at multiple levels, including authors, institutions, and research areas. The evolution of key ADNI research topics was also plotted over the studied period. Both statistical and visualization results demonstrate the increasing attention of ADNI research, strong citation impact of ADNI publications, the expanding collaboration networks among researchers, institutions and ADNI core areas, and the dynamic evolution of ADNI research topics. The visualizations presented here can help improve daily decision making based on a deep understanding of existing patterns and trends using proven and replicable data analysis and visualization methods. They have great potential to provide new insights and actionable knowledge for helping translational research in AD.


Endovascular treatment of traumatic pericallosal artery aneurysms. A case report.
Traumatic pericallosal artery aneurysms are rare complications of blunt head trauma. The pericallosal artery is torn under the sharp edge of the rigid falx. CT shows a typical hematoma in the corpus callosum. Endovascular treatment with occlusion of the aneurysm including the parent vessel with coils or glue is the best treatment option.


Aluminum as an endocrine disruptor in female Nile tilapia (Oreochromis niloticus).
The effects of aluminum on plasma ion, lipid, protein and steroid hormone concentration were evaluated in Oreochromis niloticus broodstock females. Lipid and protein concentrations from the gonads and liver were also measured. Experiments were performed at neutral and acidic water pH. Four groups of fish were tested for 96h: 1) control conditions at neutral water pH; 2) control conditions at acidic water pH (CTR-Ac); 3) aluminum at neutral water pH (Al-N); and 4) aluminum at acidic water pH (Al-Ac). Aluminum and acidic water pH exposure caused no ionoregulatory disturbances. Total lipid concentration increased in the mature gonads and decreased in the liver, suggesting an acceleration of lipid mobilization to the ovaries in animals exposed to aluminum. However, a decreased protein concentration in ovaries was also observed. Exposure of control fish to acidic water pH caused an increased concentration of plasma 17alpha-hydroxyprogesterone. However, females exposed to aluminum at acidic water pH showed a decreased of plasma 17alpha-hydroxyprogesterone and cortisol. No differences in plasma 17beta-estradiol were observed. The physiological mechanisms underlying the disturbances observed are discussed focusing on reproduction. We suggest that aluminum can be considered an endocrine disrupting compound in mature O. niloticus females.


Brain regions controlling courtship behavior in the bluehead wrasse.
Bluehead wrasses (Thalassoma bifasciatum) follow a socially controlled mechanism of sex determination. A socially dominant initial-phase (IP) female is able to transform into a new terminal-phase (TP) male if the resident TP male is no longer present. TP males display an elaborate array of courtship behaviors, including both color changes and motor behaviors. Little is known concerning the neural circuits that control male-typical courtship behaviors. This study used glutamate iontophoresis to identify regions that may be involved in courtship. Stimulation of the following brain regions elicited diverse types of color change responses, many of which appear similar to courtship color changes: the ventral telencephalon (supracommissural nucleus of the ventral telencephalon [Vs], lateral nucleus of the ventral telencephalon [Vl], ventral nucleus of the ventral telencephalon [Vv], and dorsal nucleus of the ventral telencephalon [Vd]), parts of the preoptic area (NPOmg and NPOpc), entopeduncular nucleus, habenular nucleus, and pretectal nuclei (PSi and PSm). Stimulation of two regions in the posterior thalamus (central posterior thalamic [CP] and dorsal posterior thalamic [DP]) caused movements of the pectoral fins that are similar to courtship fluttering and vibrations. Furthermore, these responses were elicited in female IP fish, indicating that circuits for sexual behaviors typical of TP males exist in females. Immunohistochemistry results revealed regions that are more active in fish that are not courting: interpeduncular nucleus, red nucleus, and ventrolateral thalamus (VL). Taken together, we propose that the telencephalic-habenular-interpeduncular pathway plays an important role in controlling and regulating courtship behaviors in TP males; in this model, in response to telencephalic input, the habenular nucleus inhibits the interpeduncular nucleus, thereby dis-inhibiting forebrain regions and promoting the expression of courtship behaviors.


Youths carrying a weapon or using a weapon in a fight: what makes the difference?
The objective of this study was to characterize weapon-carrying adolescents and to assess whether weapon carriers differ from weapon users. Data were drawn from a cross-sectional school-based survey of 7548 adolescents aged 16-20 years in Switzerland. Youths carrying a weapon were compared with those who do not. Subsequently, weapon carriers were divided into those who had used it in a fight and those who had not. Individual, family, school and social factors were analyzed using bivariate and stepwise multivariate analysis. For both genders, delinquent behavior and being victim of physical violence were associated with weapon carrying. For males, quarreling while intoxicated, being an apprentice, being sensation seekers, having a tattoo, having a poor relationship with parents and practicing unsafe sex were also related to weapon carrying. Compared with weapon carriers, female weapon users were more likely to be regular smokers. Male weapon users were foreign born, urban and apprentices; had poor school connectedness; practiced unsafe sex and quarreled while intoxicated. Carrying a weapon is a relatively frequent behavior among youths in Switzerland and a sizeable proportion of weapon carriers have used it in a fight. Weapon carrying should be part of the clinical assessment and preventive counseling of adolescents. Preventive programs specific for at-risk youth groups need to be developed.


Pain in Parkinson disease: mechanistic substrates, main classification systems, and how to make sense out of them.
Parkinson disease (PD) affects up to 2% of the general population older than 65 years and is a major cause of functional loss. Chronic pain is a common nonmotor symptom that affects up to 80% of patients with (Pw) PD both in prodromal phases and during the subsequent stages of the disease, negatively affecting patient's quality of life and function. Pain in PwPD is rather heterogeneous and may occur because of different mechanisms. Targeting motor symptoms by dopamine replacement or with neuromodulatory approaches may only partially control PD-related pain. Pain in general has been classified in PwPD according to the motor signs, pain dimensions, or pain subtypes. Recently, a new classification framework focusing on chronic pain was introduced to group different types of PD pains according to mechanistic descriptors: nociceptive, neuropathic, or neither nociceptive nor neuropathic. This is also in line with the International Classification of Disease-11 , which acknowledges the possibility of chronic secondary musculoskeletal or nociceptive pain due to disease of the CNS. In this narrative review and opinion article, a group of basic and clinical scientists revise the mechanism of pain in PD and the challenges faced when classifying it as a stepping stone to discuss an integrative view of the current classification approaches and how clinical practice can be influenced by them. Knowledge gaps to be tackled by coming classification and therapeutic efforts are presented, as well as a potential framework to address them in a patient-oriented manner.


Calcium and osmotic regulation of the Na+/H+ exchanger in neonatal ventricular myocytes.
Intracellular pH regulation in primary cultures of neonatal cardiac myocytes has been characterized. Myocytes were exposed to hyperosmolar solutions to examine the effects on pH regulation by the Na+/H+ exchanger. Exposure to 100 mM NaCl, sorbitol, N-methyl-D-glucamine, or choline chloride all caused significant increases in steady state pHi in myocytes. Omission of extracellular calcium or administration of calmodulin antagonists reduced the osmotic activation of the exchanger. The myosin light-chain inhibitor ML-7 completely blocked osmotic activation of the exchanger suggesting that myosin light-chain kinase is involved in osmotic activation of the exchanger in the myocardium. The calmodulin-dependent protein kinase II inhibitor KN-93 inhibited the rate of recovery from an acute acid load as did trifluoperazine (TFP) and the calmodulin blocker W7, [N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide]. Addition of the calcium ionophore ionomycin caused a large increase in resting pHi in isolated myocytes. However, this effect was largely resistant to HMA (5-(N,N-hexamethylene)-amiloride) indicating that an alternative mechanism of pHi regulation is responsible. The results demonstrate that the Na+/H+ exchanger of the neonatal myocardium is responsive to calcium and osmotically responsive pathways and that myosin light-chain kinase is a key protein involved in mediating the osmotic response.


Leukotrienes B4, C4, D4 and E4 in the exhaled breath condensate (EBC), blood and urine in patients with pneumoconiosis.
Leukotrienes (LTs) are involved in the pathogenesis of lung fibrosis and were increased in exhaled breath condensate (EBC) of the patients with pneumoconiosis. However the possible influence of extra-pulmonary disorders on the EBC markers is not known. Therefore in parallel with EBC, LTs' levels in the plasma and urine were measured in patients with pneumoconiosis (45 × asbestos exposure, 37 × silica exposure) and in 27 controls. Individual LTs B4, C4, D4 and E4 were measured by liquid chromatography - electrospray ionization - tandem mass spectrometry (LC-ESI-MS/MS). In EBC, LT D4 and LT E4 were increased in both groups of patients (p<0.001 and p<0.05), comparing with the controls. Both LT B4 and cysteinyl LTs were elevated in asbestos-exposed subjects (p<0.05). Asbestosis with more severe radiological signs (s1/s2-t3/u2) and lung functions impairment has shown higher cysteinyl LTs and LT C4 in the EBC (p<0.05) than mild asbestosis (s1/s0-s1/s1). In addition, in the subjects with asbestosis, cysteinyl LTs in EBC correlated with TLC (-0.313, p<0.05) and TLCO/Hb (-0.307, p<0.05), and LT C4 with TLC (-0.358, p<0.05). In pneumoconioses, EBC appears the most useful from the 3 fluids studied.


The conformation of the mucopolysaccharides. Hyaluronates.
X-ray-diffraction patterns of hyaluronate fibres from a variety of sources were obtained. Sodium hyaluronate gives well-defined patterns which index on a hexagonal unit cell with dimensions a=1.17+/-nm and a fibre repeat-distance of 2.85+/-0.03nm. A further form of sodium hyaluronate is produced by annealing at 60 degrees C in 75% relative humidity. This stable state indexes on a hexagonal unit cell of unchanged fibre repeat-distance but with a=1.87nm. The chain conformation is a threefold helix. Analysis of these diffraction patterns led to two tentative structures for sodium hyaluronate, involving different packing of the polysaccharide chains. The significance of side-chain interaction is discussed. Hyaluronic acid produces an X-ray pattern different from that obtained with the sodium salt. The fibre repeat-distance is 1.96+/-0.02nm and the unit cell appears to be monoclinic. The chain conformation is a twofold helix and conformational change between free acid and monovalent salt is discussed. These findings, together with model-building experiments, are interpreted as indicating a highly ordered structure, and the physical properties of hyaluronate solutions with regard to molecular shape and polyelectrolyte behaviour are rationalized.


[Importance of initial status of the nervous system in the developing of low intensity EMF exposure neuro-effects].
EMF neuro-effects are weak or under-threshold irritants (from the position of biological significance) and completely determined by initial status of nervous system. Initial brain bioelectrical activity characterized by the first developing symptoms of drowsiness or passive awaking condition promotes the reaction development. But emotional tension or somatic activation interfere this process. This dependence could be remedied by increasing of biological significance of low intensity EMF and changing of the analyzed physiological parameter.


Parkinson disease: pattern of functional MR imaging activation during deep brain stimulation of subthalamic nucleus--initial experience.
To prospectively determine the pattern of functional magnetic resonance (MR) imaging activation at 3 T produced by deep brain stimulation (DBS) of subthalamic nucleus (STN) for treatment of Parkinson disease and to determine the safety of DBS electrode stimulation during functional MR imaging at 3 T. Informed consent was obtained from all subjects participating in the study, and the study protocol was approved by the institutional review board at the Cleveland Clinic Foundation and was HIPAA compliant. After extensive phantom safety testing of DBS lead systems, five patients (three men, two women; mean age, 49.4 years +/- 14.5 [standard deviation]; range, 31-74 years) with percutaneously extended bilateral DBS electrodes placed in the STN for treatment of Parkinson disease were examined at 3 T on the 1st or 2nd postoperative day. Imaging consisted of a three-dimensional anatomic data set with leads disconnected and a blood oxygen level-dependent functional MR image with a single lead connected to the external pulse generator in the MR imaging control room by using stimulation parameters previously determined to produce optimal stimulation for alleviation of symptoms. A total of nine leads were tested with the functional MR imaging protocol. Subjects underwent neurologic examination immediately before and after MR imaging. All five patients completed the study without change in their neurologic examination and with activation seen in eight of nine electrodes stimulated. Activation was seen in the ipsilateral basal ganglia in all subjects and ipsilateral thalamus in six of the electrodes tested. Two of the electrode stimulations demonstrated additional activation in the STN and/or substantia nigra region adjacent to the electrode tip. For three electrode stimulations, activation was seen in the contralateral superior cerebellum. Therapeutically effective DBS of STN can be performed safely during functional MR imaging at 3 T and produces a consistent pattern of ipsilateral activation of deep brain motor structures.


Influence of different modes of real time visual information on single aimed movements.
This study shows that the difference in appearance between two types of computer generated real time visual information namely, cursor and trace feedback, becomes visible in the execution of aimed stylus movements. Movement execution was scrutinized by division into four relevant subsequent constituents on the basis of key kinematic events. As expected, the final section of a movement took more time and showed a larger number of adjustments in the cursor condition. Instead, the influence of trace feedback on movement execution became discernible in earlier sections of movement execution. When a combination of feedback types was employed, trace feedback seemed to dominate the impact on execution. Discussion focuses on the contribution of the present study to understanding the impact of specific features inherent to types of real time visual feedback, and the relevance of the present study's result to future research.


[Patients with Korsakoff's syndrome in a nursing home: characteristics and comorbidity].
This article presents characteristics and co-morbidity of patients with Korsakoff's syndrome after admission to a nursing home. The medical charts were studied of all patients with Korsakoff's syndrome, admitted between 1984 and 1998 to the special ward for Korsakoff patients of a nursing home in the eastern part of the Netherlands. The 'Standard of classification for diseases in nursing homes' ('Standaard van classificatie voor ziekten in de verpleeghuisgeneeskunde') was used to classify co-morbidity. The group included 77 patients. Almost 75% were male. The average age was 53 years on admission. More than 50% of the patients were divorced. After admission, 30% of the group was transferred to a different setting. The estimated average follow-up period was 7.1 year. Patients had 2.9 co-morbid conditions at admission. Diseases due to alcoholism were frequently diagnosed. During the stay skin diseases, psychological disorders and behavioural disorders were frequently noticed. The prevalence of cognitive impairment and alcohol dementia was surprisingly high. Thirteen patients died, most of them of cancer or combined cardiovascular and respiratory diseases. We conclude that patients with Korsakoff's syndrome are an unexplored area in nursing homes. This group of nursing home inhabitants shows its own characteristics and co-morbidity.


[The altitude chamber flights as a method of the psychophysiologic training of pilots against stress situations].
The study of literature and aviation incidents during the depressurization of the aircraft cabin in the altitude higher than 12 km. Shows that their serious outcomes are often connected with wrong evaluation of the emergency situation by pilots, making incorrect decision and carrying out inadequate actions. In order to improve psychophysiologic training for stratosphere flights and to increase their safety the authors suggest approbated methods of periodic altitude chamber flights of flying personnel to the altitude of 13.5 km., imitating the stress situation during the depressurization of the cabin and enabling to form adequate algorithm of pilots' behaviour in these conditions.


Microencapsulation to reduce mechanical loss of microspheres: implications in myocardial cell therapy.
Previous regenerative studies have demonstrated massive cell losses after intramyocardial cellular delivery. Therefore, efforts at reducing mechanical losses may prove more successful in optimising cellular therapy. In this study, we hypothesized that escalating mesenchymal stem cells (MSCs) dose will not produce corresponding improvement in cardiac function due to washout of the small cells in microcirculation. Using microspheres similar in size to MSCs, that are encapsulated in alginate-poly-l-lysine-alginate (APA), we tested the hypothesis that size is an important factor in early losses. In experiment I, five groups of rats (n=9 each) underwent coronary ligation; group I had no treatment; the other groups received escalating 0.5 × 10(6), 1.5 × 10(6), 3 × 10(6) and 5 × 10(6) of MSCs each. Echocardiogram was performed at baseline, 2 days and 7 weeks after surgery. In experiment II, cell-sized microspheres (10 μm) were encapsulated in APA microcapsules. In group I (n=16), rats received bare microspheres, group II (n=16) microspheres within 200 μm microcapsules and in group III (n=16), microspheres within 400 μm microcapsules. After 20 min, hearts were quantified for the amount retained. Myocardial function did not improve further with escalating cell doses beyond an initial response at 1.5 × 10(6) cells. Encapsulated microspheres in 200 μm and 400 μm microcapsules demonstrated a fourfold increase in retention rate compared with 10 μm microspheres. We concluded that suboptimal functional improvement in this animal model starts at 1.5 × 10(6) cells and does not respond to escalating cell doses. Improving mechanical retention is possible by increasing the size of the injectate. Microencapsulation could be used to encapsulate donor cells and facilitate functional improvement in cellular heart failure therapy.


Primary structure of potato kunitz-type serine proteinase inhibitor.
The serine proteinase inhibitor (PSPI-21) isolated from potato tubers (Solanum tuberosum L.) comprises two protein species with pI 5.2 and 6.3, denoted as PSPI-21-5.2 and PSPI-21-6.3, respectively. They were separated by anion exchange chromatography on a Mono Q FPLC column. Both species tightly inhibit human leukocyte elastase, whereas their interaction with trypsin and chymotrypsin is substantially weaker. The sequences of both PSPI-21-5.2 and PSPI-21-6.3 were determined by analysis of overlapping peptides obtained from the oxidized or reduced and S-pyridylethylated proteins after digestion with trypsin or pepsin. Both species of PSPI-21 are composed of two chains, named chains A and B, which are linked by a disulfide bridge between Cys(146) and Cys(157). The other disulfide bridge is located within the A chains between Cys(48) and Cys(97). The amino acid sequences of the large A chains of the two forms, consisting of 150 amino acids residues each, differ in a single residue at position 52. The small chains B, containing 37 and 36 residues in PSPI-21-6.3 and PSPI-21-5.2, respectively, have nine different residues. The entire amino acid sequences of the two inhibitors show a high degree of homology to the other Kunitz-type proteinase inhibitors from plants.


Taking a closer look: On the operation of nonconscious impression formation.
In this article we analyzed the information processing that underlies nonconscious impression formation. In the first experiment, the nonconscious activation of the impression formation goal led to a faster analysis of the trait implications of behaviors, compared with a control group. In Experiment 2, participants who were nonconsciously primed with an impression formation goal were more likely than those in a control condition to form associations in memory between behaviors and implied traits. In Experiment 3, nonconsciously primed participants were more sensitive than those in a control condition to whether inconsistent trait information was relevant or irrelevant to the actor's disposition. Moreover, in Experiments 2 and 3, those with a nonconscious goal showed just as much evidence of impression formation as those who were consciously and intentionally trying to form an impression. Implications for nonconscious goal pursuit and impression formation are discussed.


Biodegradation property of beta-tricalcium phosphate-collagen composite in accordance with bone formation: a comparative study with Bio-Oss Collagen® in a rat critical-size defect model.
The objective of this study was to compare osteoconductivity and biodegradation properties of an in-house fabricated beta-tricalcium phosphate (b-TCP)-collagen composite with those of Bio-Oss Collagen® (Osteohealth, Shirley, NY, USA) using a rat calvarial critical-size defect model. b-TCP-collagen composite material was fabricated by mixing b-TCP granules having a particle size of 0.15 to 0.8 mm and 75% porosity, with bovine dermis-derived soluble collagen sponge. The dry weight ratio of b-TCP granules-to-collagen ratios was 4:1. Bio-Oss Collagen or the b-TCP-collagen composite was used to fill a 5.0 mm-diameter calvarial defect in rats. The defects were evaluated by histological and histomorphological analyses of decalcified histological sections with hematoxylin and eosin staining 6 and 10 weeks, respectively, after surgery. The defect implanted with the b-TCP composite contained immature bone structures with dense connective tissue in contrast to the abundant fibrous tissue, but no trabecular structure was observed within the defect implanted with Bio-Oss Collagen at 6 weeks postoperatively. Eventually, the defect filled with the b-TCP composite was covered with dense, continuous, mature bone tissue with complete replacement of the graft material. However, in defects filled with Bio-Oss Collagen, only dense connective tissue, containing limited amounts of immature trabecular bone and abundant remnant Bio-Oss particles, was observed. Histomorphological analysis revealed that the b-TCP composite caused greater tissue augmentation with a larger volume of bone tissue observed in the defect and greater bioabsorption of remnant material than Bio-Oss Collagen. These results indicated that the b-TCP composite has greater osteoconductivity and better biodegradation properties than Bio-Oss Collagen; these properties of the b-TCP-collagen composite complimented bone formation and remodeling.


Attention Deficit Hyperactivity Disorder (ADHD) and its Comorbid Mental Disorders: An Evaluation of their Labor Market Outcomes.
Attention-deficit/hyper-activity disorder's (ADHD) prevalence rate has been increasing during the last decade. Evidence from different studies suggests that the effect of ADHD on earnings and employment could be more detrimental than other disorders such as depression or anxiety. Although it is widely known that these mental disorders can coexist with ADHD, none of these studies has considered the joint evaluation of ADHD and its comorbidities. In this paper, we evaluate whether ADHD is a more severe disorder than three other comorbid mental disorders -- learning disabilities, depression, and anxiety -- regarding their effects on earnings and employment. We use the National Longitudinal Study of Adolescent to Adult Health to estimate regression models of earnings and employment. We consider the use of the sampling weights, school fixed effects, and multiple imputation of missing values. Robustness checks include a more exogenous measure of depression. On average, an individual with ADHD or depression (but not both) has around 20% lower earnings than those without any of these disorders. The earnings gap for learning disabilities is around 25%. The probability of being employed is 5 percentage points lower for an individual who has any of these disorders. Anxiety is not associated with lower earnings. Females with learning disabilities have an earnings gap that is 20 percentage points larger than males. The employment gap for females or African Americans with learning disabilities is 8%. In contrast to existing evidence, we find that ADHD is not more serious than learning disabilities or depression. Although the magnitude of each employment gap is not substantial, the fact that these are comorbid disorders indicates that their joint occurrence could be detrimental for employment. Females and African Americans with learning disabilities could face more adverse labor market outcomes. Even though our results are robust to a more exogenous measure of depression, the potential endogeneity of the diagnosis of ADHD or anxiety could still bias the estimates. However, baseline results are also robust when individuals with an age of diagnosis equal to the year of the interview are dropped from the estimations. The increasing number of Supplemental Security Income beneficiaries with ADHD is in line with the observed tendency to screen for ADHD more frequently than for other comorbid disorders, and with the evidence of ADHD overdiagnosis. Since our study shows that other disorders are equally or potentially more disabling than ADHD in terms of labor market outcomes, mental health policies should be reoriented to prevent policies targeting ADHD from crowding out those for other mental disorders. It would be interesting to verify our findings using a more recent cohort with a higher rate of ADHD diagnosis. Since we show that there are unexplained gender and race differences that persist after controlling for educational attainment or the number of hours worked, more research is needed to determine the reason behind these differences. Further research about the positive school effect on the adulthood performance of ADHD children is needed.


Definitions of sleeplessness in children with attention-deficit hyperactivity disorder (ADHD): implications for mothers' mental state, daytime sleepiness and sleep-related cognitions.
Sleep disturbances are common in children with attention-deficit hyperactivity disorder (ADHD). Sleeplessness is frequently reported although results are inconsistent perhaps because different definitions for it are applied. This study looked at maternal functioning and child objective sleep patterns in relation to different definitions of sleeplessness in children with ADHD. The study included 45 children (aged 3-14 years) with ADHD and their mothers. Sleeplessness was defined according to: (i) yes/no report of whether mothers thought their children had a problem with sleeplessness (Maternal definition MD) and (ii) mothers' responses to a quantitative standardized questionnaire (Quantitative definition QD) designed to detect the frequency and duration of parent-reported problems with settling, night waking and early waking. Objective sleep patterns were also assessed by means of actigraphy. Maternal mental health, daytime sleepiness and cognitions related to child sleep were assessed by questionnaire. Both definitions appeared to tap similar although slightly different constructs. There were no group differences in objective sleep patterns. Maternal mental health was found to be significantly worse in the mothers who considered their child to be sleepless (MD) (P < 0.025), but not in those mothers whose child was found to be sleepless according to the standardized criteria (QD). Maternal sleepiness did not differ between groups. For both categories of sleepless children (MD and QD), the mothers had significantly more doubts about their competency as a parent (P < 0.01 and P < 0.025, respectively) compared to mothers of children without sleeplessness. Two different maternal assessments of child sleeplessness in children with ADHD may assess subtly different constructs, but both may provide useful information about potential problems across the family.


The many issues in reliability research: choosing from a horn of plenty.
We respond to three commentaries on our discussion article on different conceptions of test score reliability. First, we discuss the use of standard errors for reliability estimates. Second, we discuss the desirability not to confuse issues pertaining to the dimensionality of the test data (closely related to construct validity) and the degree to which measurement values are repeatable under the same circumstances (i.e., the reliability issue). Third, we discuss a new reliability estimation method that is almost unbiased irrespective of the dimensionality of the test data.


Proteomics in Hypothermia as Adjunctive Therapy in Patients with ST-Segment Elevation Myocardial Infarction: A CHILL-MI Substudy.
Cardiovascular and inflammatory biomarkers in therapeutic hypothermia have been studied in cardiac arrest, but data on patients with ST-segment elevation myocardial infarction (STEMI) treated with therapeutic hypothermia are currently unavailable. A multiplex proximity extension assay allowed us to measure 157 cardiovascular disease (CVD) and inflammatory disease-related biomarkers in patients from the international, multicenter, and randomized trial; CHILL-myocardial infarction (MI) and to explore the associations of cardiovascular and inflammatory biomarkers. Blood samples were obtained from 119 patients with STEMI, randomized to hypothermia as adjunctive therapy to percutaneous coronary intervention (PCI) or standard care with PCI only. Blood samples were obtained at baseline (0 hour), 6, 24, and 96 hours post PCI, and stored at -80°C until they were analyzed by PROSEEK Multiplex CVD and PROSEEK Multiplex INF (Olink Bioscience, Uppsala, Sweden). Peak values from 6, 24, and 96 hours postrandomization were compared between treatment groups. One hundred fifty-seven cardiovascular and inflammatory biomarkers were evaluated. Peak values of four biomarkers (BDNF, DNER, CCL20, MMP3) were reduced in the hypothermia group as compared with the control group. In addition, seven markers were slightly elevated in the hypothermia group (OPG, FGF21, FS, IL12B, PRL, TIM, IL6). In a prespecified subgroup analysis of anterior infarctions, two additional markers were reduced (PTX3 and SELE). In this explorative proteomic study from the randomized trial CHILL-MI, four biomarkers were identified as having reduced peak plasma values in patients with STEMI treated with therapeutic hypothermia as adjunctive therapy to PCI as compared with patients treated with standard care of PCI. In addition, seven biomarkers were elevated in the group treated with hypothermia therapy. The effect of hypothermia on biomarker peak values was modest, possibly due to a low reduction in mean body temperature. Whether a faster and deeper cooling results in more pronounced effects is yet to be established.


[Seasonal dynamics of the content of proteins inhibiting trypsin-like proteases in the leaves of common oak (Quercus petraea Liebl.)].
After the successive purification of proteins-inhibitors of trypsin-like proteases using the methods of chromatography on DEAE-cellulose, trypsin-agarose and sephadex G-50, their numerous forms with the values of molecular weight 3.5; 6.3; 12.5; and 25.0 kDa have been found out. From spring to autumn the increase of the absolute content of proteins-inhibitors of all types was accompanied by the change of their relative content. The more than by an order increase of general content of proteins inhibitors is discussed from the position of their stress origin as a result of the effect of unfavorable ecological factors.


Petrobactin is produced by both pathogenic and non-pathogenic isolates of the Bacillus cereus group of bacteria.
Petrobactin is the primary siderophore synthesized by Bacillus anthracis str Sterne and is required for virulence of this organism in a mouse model. The siderophore's biosynthetic machinery was recently defined and gene homologues of this operon exist in several other Bacillus strains known to be mammalian pathogens, but are absent in several known to be harmless such as B. subtilis and B. lichenformis. Thus, a common hypothesis regarding siderophore production in Bacillus species is that petrobactin production is exclusive to pathogenic isolates. In order to test this hypothesis, siderophores produced by 106 strains of an in-house library of the Bacillus cereus sensu lato group were isolated and identified using a MALDI-TOF-MS assay. Strains were selected from a previously defined phylogenetic tree of this group in order to include both known pathogens and innocuous strains. Petrobactin is produced by pathogenic strains and innocuous isolates alike, and thus is not itself indicative of virulence.


Bismahanine exerts anticancer effects on human cervical cancer cells by inhibition of growth, migration and invasion via suppression of NF-kB signalling pathway.
Bismahanine, a carbazole alkaloid, has been shown to exhibit tremendous pharmacological potential. In this study, the effect Bismahanine was examined against human cervical cancer cells. The HeLa human cervical cancer cells were treated with various concentrations of Bismahanine from 0-320 μM for 24 h. The anticancer effects of Bismahanine were measured by WST-1 cell viability assay. DAPI and annexin V/propidium iodide (PI) assays were employed to examine the induction of apoptosis. Transwell assays were performed to examine the cell migration and invasion. The expression of the proteins was examined by western blot analysis. Bismahanine decreased the viability of HeLa cells and exhibited an IC50 of 20 µM due to induction of apoptosis as indicated by DAPI staining. Additionally, the annexin V/PI staining revealed that apoptotic cell percentage increased with increasing concentration of Bismahanine. The expression of Bcl-2 was decreased while that of Bax, Caspase 3 and 9 was increased. Bismahanine treatment also resulted in significant decrease of metalloproteinase (MMP) 2 , 3 and 9 expressions. Transwell assays showed that Bismahanine inhibited the migration and invasion of HeLa cells. Bismahanine exhibits significant anti-proliferative effects on the cervical cancer cells and may prove essential in the development of chemotherapy for cervical cancer.


Specific impairments of selective attention in mild Alzheimer's disease.
The aim of the present study was to evaluate selective visual attention in subjects affected by Alzheimer's Disease (AD), by means of a computerized spatial exploration test that adopts a Touch Screen (TS) interface, which has already proved able to characterize alternative strategies in performing search tasks. We assessed a group of 16 patients affected by mild to moderate AD, comparing them with 16 control subjects matched for age and education. In the experimental tasks the performance of the AD patients was worse than that of the normal elderly, both quantitatively (slower speeds) and qualitatively (poorer planning and higher number of omissions and perseverations). In the visual attention tasks there appeared to be no close connection between AD patients' performance and increased Reaction Times (RT); this evidenced a specific role of non-elementary cognitive structures enclosed in a higher attentional domain, rather than a general decrease in the speed of basic cognitive processes. Our results are in line with specific AD literature: while psychomotor speed and lower attention levels (sensorimotor) are preferentially impaired in subcortical forms of dementia, the higher levels of selective and divided attention could be the first to deteriorate and appear more markedly disrupted in the Alzheimer type of dementia.


De novo design and synthesis of a 30-cistron translation-factor module.
Two of the many goals of synthetic biology are synthesizing large biochemical systems and simplifying their assembly. While several genes have been assembled together by modular idempotent cloning, it is unclear if such simplified strategies scale to very large constructs for expression and purification of whole pathways. Here we synthesize from oligodeoxyribonucleotides a completely de-novo-designed, 58-kb multigene DNA. This BioBrick plasmid insert encodes 30 of the 31 translation factors of the PURE translation system, each His-tagged and in separate transcription cistrons. Dividing the insert between three high-copy expression plasmids enables the bulk purification of the aminoacyl-tRNA synthetases and translation factors necessary for affordable, scalable reconstitution of an in vitro transcription and translation system, PURE 3.0.


Quantitative real-time polymerase chain reaction as an efficient molecular tool for detecting minimal residual disease in Moroccan chronic myeloid leukemia patients.
Chronic myeloid leukemia (CML) is characterized by BCR-ABL translocation and an increased number and migration of immature myeloid cells into the peripheral blood. The detection limit of the BCR-ABL transcript, particularly after treatment, is controversial. In the present study, we used quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) to monitor BCR-ABL expression in Moroccan CML patients undergoing imatinib treatment, and compared the results with those of conventional PCR and fluorescence in situ hybridization (FISH). The aim of this study was to establish a new molecular tool for in vitro diagnosis of CML. In a retrospective comparative analysis, 20 CML Moroccan patients who had received imatinib treatment (N = 20) were analyzed by real-time PCR, conventional RT-PCR, and FISH. Half of the samples analyzed (N = 10) were positive for BCR-ABL gene expression, while the other half (N = 10) were negative according to conventional PCR. Interestingly, 5 of the 10 samples shown to be negative by conventional PCR showed positive expression of the BCR-ABL gene according to RT-qPCR. The RT-qPCR results were confirmed by FISH, which revealed a high concordance (100%) rate. We found that real-time RT-qPCR is more reliable and should be used in Moroccan biomedical analysis laboratories to monitor CML progression, particularly for minimal residual disease, following imatinib treatment.


Local and long-range reciprocal regulation of cAMP and cGMP in axon/dendrite formation.
Cytosolic cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) often mediate antagonistic cellular actions of extracellular factors, from the regulation of ion channels to cell volume control and axon guidance. We found that localized cAMP and cGMP activities in undifferentiated neurites of cultured hippocampal neurons promote and suppress axon formation, respectively, and exert opposite effects on dendrite formation. Fluorescence resonance energy transfer imaging showed that alterations of the amount of cAMP resulted in opposite changes in the amount of cGMP, and vice versa, through the activation of specific phosphodiesterases and protein kinases. Local elevation of cAMP in one neurite resulted in cAMP reduction in all other neurites of the same neuron. Thus, local and long-range reciprocal regulation of cAMP and cGMP together ensures coordinated development of one axon and multiple dendrites.


A statistical analysis of the effect of warfare on the human secondary sex ratio.
Many factors have been hypothesized to affect the human secondary sex ratio (the annual percentage of males among all live births), among them race, parental ages, and birth order. Some authors have even proposed warfare as a factor influencing live birth sex ratios. The hypothesis that during and shortly after periods of war the human secondary sex ratio is higher has received little statistical treatment. In this paper we evaluate the war hypothesis using 3 statistical methods: linear regression, randomization, and time-series analysis. Live birth data from 10 different countries were included. Although we cannot speak of a general phenomenon, statistical evidence for an association between warfare and live birth sex ratio was found for several countries. Regression and randomization test results were in agreement. Time-series analysis showed that most human sex-ratio time series can be described by a common model. The results obtained using intervention models differed somewhat from results obtained by regression methods.


Relationship of white potato to other vegetable consumption by schoolchildren and adolescents in the U.S.A: National Health and Nutrition Examination Survey, 2003-2008.
Intake of white potatoes in and out of school was estimated to provide context for a recent proposal by the U.S. Department of Agriculture to limit provision of white potatoes in U.S. school meals. Mean daily servings of white potatoes and other vegetables consumed in and out of school for school-aged children were estimated from two days of 24 h dietary recall data from the National Health and Nutrition Examination Surveys (NHANES). Total energy intake and percentage energy contribution from discretionary oils and solid fats were also estimated for all white potato dishes consumed. The NHANES is nationally representative of the U.S. civilian noninstitutionalized population. Children and adolescents aged 6–19 years (n 8466) from three combined NHANES cycles (2003–2004, 2005–2006 and 2007–2008). White potatoes represented 32% of all vegetable servings consumed by U.S. children and adolescents. Preparations high in fats and oils, including French fries, were most popular both in and out of school. Mean consumption of white potatoes obtained from school cafeterias was approximately 0.05 servings/d among all children and adolescents, and about 0.15 servings/d among children and adolescents acquiring at least one item from the school cafeteria, implying current weekly intake levels well below the limit of 2 servings/week proposed by the U.S. Department of Agriculture. Although white potatoes represent a substantial proportion of vegetable consumption among school-aged children, it is unclear that proposed limits would influence white potato intake from school cafeterias. Policy makers should consider targeting preparation methods to improve the healthfulness of white potato dishes.


Hot-spot engineering in polygonal nanofinger assemblies for surface enhanced Raman spectroscopy.
Multiparticle assemblies of nanoscale structures are the fundamental building blocks for powerful plasmonic devices. Here we show the controlled formation of polygonal metal nanostructure assemblies, including digon, trigon, tetragon, pentagon, and hexagon arrays, which were formed on top of predefined flexible polymer pillars that undergo self-coalescence, analogous to finger closing, with the aid of microcapillary forces. This hybrid approach of combining top-down fabrication with self-assembly enables the formation of complex nanoplasmonic structures with sub-nanometer gaps between gold nanoparticles. On comparison of the polygon-shaped assemblies, the symmetry dependence of the nanoplasmonic structures was determined for application to surface enhanced Raman spectroscopy (SERS), with the pentagonal assembly having the largest Raman enhancement for the tested molecules. Electromagnetic simulations of the polygonal structures were performed to visualize the field enhancements of the hot spots so as to guide the rational design of optimal SERS structures.


Colocalization of chaperone Cpn60, proinsulin and convertase PC1 within immature secretory granules of insulin-secreting cells suggests a role for Cpn60 in insulin processing.
Many of the mechanisms that control insulin processing and packaging by interaction with different elements along the secretory pathway remain poorly understood. We have investigated the possibility that Cpn60, a member of the heat shock protein family, may be present in rat insulin-secreting cells, participating in the proinsulin-insulin maturation process. Immunofluorescence and high resolution immunocytochemical studies revealed the presence of the Cpn60 protein all along the insulin secretory pathway, being particularly abundant over the proinsulin-containing immature secretory granules. Double-labeling experiments showed associations between Cpn60 and proinsulin, as well as between Cpn60 and PC1 convertase, with a preferential binding to proinsulin. These findings paralleled those of coimmunoprecipitation studies showing the Cpn60 chaperone and the mature form of the PC1 convertase in proinsulin immunoprecipitates, as well as the PC1 in Cpn60 immunoprecipitates from total islet cell extracts. In vitro binding of Cpn60 to proinsulin, insulin and glucagon was also documented. Cpn60, significantly abundant in proinsulin-containing secretory granules where conversion of proinsulin to insulin takes place, and the colocalization of the chaperone with proinsulin and PC1 convertase suggest that the Cpn60 protein may play a role directing precise molecular interactions during insulin processing and/or packaging.


Nutrient intake values for folate during pregnancy and lactation vary widely around the world.
Folate is a B-vitamin with particular importance during reproduction due to its role in the synthesis and maintenance of DNA. Folate is well known for its role in preventing neural tube defects (NTDs) during the periconceptional period. There is also an increased need for folate throughout pregnancy to support optimal growth and development of the fetus and blood volume expansion and tissue growth of the mother. During lactation, women are at risk of folate deficiency due to increased demands to accommodate milk folate levels. Nutrient Intake Values (NIVs) for folate have been calculated to take into account additional needs during pregnancy and lactation. However, these values vary widely between countries. For example, the folate requirement that is set to meet the needs of almost all healthy women during pregnancy varies from 300 µg/day in the United Kingdom to 750 µg/day in Mexico. Currently, there is no accepted standardized terminology or framework for establishing NIVs. This article reviews country-specific NIVs for folate during pregnancy and lactation and the basis for setting these reference values.


The IGF system and longitudinal growth in preterm infants in relation to gestational age, birth weight and gender.
Growth factors in the blood of very preterm infants may reflect growth and contribute to the understanding of early development. We investigated postnatal levels of insulin-like growth factors (IGFs) in infants born very preterm and related them to early growth development. Blood samples were analyzed weekly for IGF-I, IGF-II, IGF binding protein (BP)-1, IGFBP-3, and acid-label subunit (ALS). 73 children born very preterm (gestational age (GA) <32 weeks) were divided according to their gender-specific birth weight standard deviation score (SDS) into either appropriate for GA (AGA) or small for GA (SGA). Fifty-two (71%) and forty-three (59%) infants completed follow-up with anthropometry at approximately 3 years and at 5 years of age respectively. Thirty-six subjects (49%) had blood sampling for IGF-I and IGFBP-3 measurements up to 3 years of age. IGF-I, IGFBP-3, and ALS levels increased in all groups from week 31 to week 36, with generally lower levels in the SGAs, with a concomitant lower growth velocity. Postnatal ALS was strongly associated with IGF-I and IGFBP-3 in boys, girls and AGA infants. IGF-II was higher in earlier born preterms (GA < 27 weeks) at postmenstrual ages 27.5-29.9 weeks compared with SGAs and late GA (GA ≥ 27 weeks) preterms (p < .0001). IGF-II, in contrast to IGF-I, did not differ between SGAs and AGAs at weeks 31-36. Mean IGFBP-1 was highest in the SGAs compared to AGAs at mean week 28,5 and 31 (p = .001) and IGFBP-1 levels were elevated in relation to IGF-I in the SGAs at that period. At follow-up, the increase in IGF-I between week 31 and 33.5 was a significant positive determinant of height SDS at 3 and 5 years of age in forward multiple regression analysis, independent of target height. This is the first study to investigate postnatal ALS levels in preterm infants. In very preterm infants, IGF-II is less affected by size at birth during early postnatal weeks compared with IGF-I. Early elevated IGFBP-1 might protect the SGA infants from an intense metabolic rate. Our results indicate that anabolic and metabolic processes during weeks 31-36 predicts later height.


Patient and public attitudes to and awareness of clinical practice guidelines: a systematic review with thematic and narrative syntheses.
Clinical practice guidelines are typically written for healthcare providers but there is increasing interest in producing versions for the public, patients and carers. The main objective of this review is to identify and synthesise evidence of the public's attitudes towards clinical practice guidelines and evidence-based recommendations written for providers or the public, together with their awareness of guidelines. We included quantitative and qualitative studies of any design reporting on public, patient (and their carers) attitudes and awareness of guidelines written for providers or patients/public. We searched electronic databases including MEDLINE, PSYCHINFO, ERIC, ASSIA and the Cochrane Library from 2000 to 2012. We also searched relevant websites, reviewed citations and contacted experts in the field. At least two authors independently screened, abstracted data and assessed the quality of studies. We conducted a thematic analysis of first and second order themes and performed a separate narrative synthesis of patient and public awareness of guidelines. We reviewed 5415 records and included 26 studies (10 qualitative studies, 13 cross sectional and 3 randomised controlled trials) involving 24 887 individuals. Studies were mostly good to fair quality. The thematic analysis resulted in four overarching themes: Applicability of guidelines; Purpose of guidelines for patient; Purpose of guidelines for health care system and physician; and Properties of guidelines. Overall, participants had mixed attitudes towards guidelines; some participants found them empowering but many saw them as a way of rationing care. Patients were also concerned that the information may not apply to their own health care situations. Awareness of guidelines ranged from 0-79%, with greater awareness in participants surveyed on national guideline websites. There are many factors, not only formatting, that may affect the uptake and use of guideline-derived material by the public. Producers need to make clear how the information is relevant to the reader and how it can be used to make healthcare improvements although there were problems with data quality. Awareness of guidelines is generally low and guideline producers cannot assume that the public has a more positive perception of their material than of alternative sources of health information.


Effect of cellular senescence on the P2Y-receptor mediated calcium response in trabecular meshwork cells.
The objectives of this study were to evaluate the potential release of ATP that is mediated by mechanical stress on trabecular meshwork (TM) cells, to identify the specific P2Y receptors mediating the ATP response, and to determine whether cellular senescence might interfere with the P2Y receptor-mediated calcium response, thus contributing to the loss of physiologic TM function in aging and primary open angle glaucoma (POAG). Experiments were conducted using primary cultures of porcine TM cells. Cyclic mechanical stretch (10% stretching/second) was generated using the Flexcell system. ATP release and ectoATPase activity induced by mechanical stress were measured using a luciferin/luciferase assay. Replicative senescence was induced by passing the cells 18 times at a 1:2 split ratio and confirmed by the presence of senescence-associated beta-galactosidase (sa-beta-gal) and autofluorescence. For calcium imaging, cells were plated on gelatin-coated coverslips, bathed in calcium Ringer's solution, and loaded with fluo-4 (5 microM) for 1 h. Agonists of P2Y1 (ADP) and P2Y2/P2Y4 (ATP, UTP) receptors at 10 microM or 100 microM concentrations were added to the bathing medium. Relative changes in cytosolic calcium concentration as a function of time were measured by fluorescent microscopy and reported as peak amplitudes of fluo-4 fluorescence normalized to baseline values (deltaF/Fo). Mechanical stress induced an increase in ATP release from TM cells (258%+/-23% at 15 min, 188%+/-11% at 30 min, and 900%+/-203% at 1 h; p<0.017, n=4) as well as an increase in ectoATPase activity present in the extracellular media during the first 15 min of stress (57%+/-15%, p=0.011, n=4). The P2Y receptor agonists listed above induced a concentration-dependent rise in intracellular calcium in the TM cells. The peak amplitude, deltaF/Fo, was 1.07+/-0.12 (n=3) for 10 microM ADP, 2.59+/-0.33 (n=6) for 100 microM ADP, 1.21+/-0.64 (n=12) for 10 microM UTP, 3.22+/-2.0 (n=12) for 100 microM UTP, 0.88+/-0.40 (n=9) for 10 microM ATP, and 1.37+/-0.61 (n=25) for 100 microM ATP. Cells at passage 18 showed significantly lower levels of intracellular calcium induced by ATP (36%), UTP (34%), and ADP (52%) compared to cells at passage 2, independent from any changes in P2Y receptor changes in expression. The ability to release ATP in response to mechanical stress and the presence of functional P2Y receptors in TM cells suggest a novel mechanism by which TM cells could sense and respond to changes in intraocular pressure (IOP). In addition, the decrease in P2Y receptor-mediated calcium responses observed in senescent TM cells suggests that the disregulation of calcium homeostasis in senescence may contribute to the alterations of the TM in aging and POAG.


Effect-Directed Analysis of Human Peroxisome Proliferator-Activated Nuclear Receptors (PPARγ1) Ligands in Indoor Dust.
Agonism of human peroxisome proliferator-activated nuclear receptor gamma (PPARγ1) was recently observed in 15 of 25 samples of indoor dust extracts at environmentally relevant exposure levels. In this study, an effect-directed analysis approach was used to identify the primary contributors of PPARγ1 activity in the dust extracts. Three dust extracts showing significant PPARγ1 activity were fractionated with normal phase high-performance liquid chromatography (NP-HPLC) and each fraction was tested for PPARγ1 activity. Three dust extracts showed a similar PPARγ1 activity distribution in the NP-HPLC fractions. In most active fractions, fatty acids (FAs), including oleic acid, stearic acid, palmitic acid and myristic acid, were the primary chemicals identified using gas-chromatography mass spectrometry (GC-MS). Chemical measurements of the FAs in house dust extracts revealed a positive and significant correlation with the observed PPARγ1 activity. To test the role of FAs in the activity, a mixture of four FAs was prepared in the ratios measured in the dust samples and tested for activity. The activity of this mixture was 30-50% of the activity observed in the dust extracts, suggesting they were contributing to the observed activity, but also suggesting additional unknown compounds are likely still present in the dust extracts. To tentatively identify sources of FAs in the dust samples, FAs were quantified in human/animal hair, dead skin cells, and cooking oil. FAs were abundant in all samples and our data indicate that all of these may be sources to indoor dust.


TcpC secreting uropathogenic E. coli promoted kidney cells to secrete MIP-2 via p38 MAPK pathway.
Pyelonephritis is an infection of the upper urinary tract with characteristic histological change to neutrophil infiltration in the kidney. The majority of pyelonephritis is caused by uropathogenic Escherichia (E.) coli (UPEC) bearing distinct virulence factors. Toll/interleukin‑1 receptor domain‑containing protein C (TcpC) encoded by E. coli is an important virulence factor in the majority of strains of UPEC and inhibits macrophage‑mediated innate immunity, which serves an essential role in the pathogenesis of pyelonephritis. In the present study, it was demonstrated that TcpC induced kidney cells to produce macrophage inflammatory protein‑2 (MIP‑2; also known as C‑X‑C motif chemokine 2). MIP‑2 concentration in kidney homogenates from TcpC‑secreting UPEC CFT073 (TcpCwt) murine pyelonephritis models was significantly higher compared with that in kidney homogenates from tcpC knockout CFT073 (TcpC‑/‑) models. In vitro, TcpCwt dose‑dependently promoted MIP‑2 secretion in HEK‑293 cells. The concentration of MIP‑2 in culture supernatants of HEK‑293 co‑cultured with TcpCwt was profoundly higher compared with that of HEK‑293 co‑cultured with TcpC‑/‑. In the presence of anti‑TcpC antibody, the enhancement effect of TcpCwt on MIP‑2 production was completely abrogated, suggesting that the enhanced production of MIP‑2 was mediated by secreted TcpC. Furthermore, it was demonstrated that TcpC‑/‑ treatment had no effect on the p38 mitogen activated protein kinase (MAPK) signaling pathway, while TcpCwt treatment resulted in the activation of p38 MAPK in HEK‑293 cells, as indicated by a simultaneous increase in p38 and phosphorylated‑p38. In addition, inhibition of p38 MAPK with SB203580 significantly decreased MIP‑2 concentration and neutrophil recruitment activity in the supernatants of HEK‑293 cells co‑cultured with TcpCwt. This indicates that TcpC may promote MIP‑2 production in kidney cells through the p38 MAPK signaling pathway. Taken together, the data of the present study demonstrated that TcpC can induce MIP‑2 production, which may contribute to the characteristic histological change associated with pyelonephritis. This data has provided novel evidence to further clarify the pathogenesis of pyelonephritis and novel directions on the pathogenicity of TcpC‑secreting UPEC.