Inter-tissue differences in fatty acid incorporation as a result of dietary oil manipulation in Port Jackson sharks (Heterodontus portusjacksoni).
Fatty acid profile analysis is a tool for dietary investigation that may complement traditional stomach contents analysis. While recent studies have shown that the liver of sharks fed different diets have differing fatty acid profiles, the degree to which diet is reflected in shark blood serum and muscle tissue is still poorly understood. An 18-week controlled feeding experiment was undertaken using captive Port Jackson sharks (Heterodontus portusjacksoni). Sharks were fed exclusive diets of artificial pellets treated with fish or poultry oil and sampled every 6 weeks. The fatty acid profiles from liver, blood serum, and muscle were affected differently, with the period from which significant differences were observed varying by tissue and diet type. The total fatty acid profiles of fish oil and poultry oil fed sharks were significantly different from week 12 onwards in the liver and blood serum, but significant differences were only observed by week 18 in the muscle tissue of sharks fed different diets. The drivers of dissimilarity which aligned with dietary input were 14:0, 18:2n-6, 20:5n-3, 18:1n-9 and 22:6n-3 in the liver and blood serum. Dietary fatty acids accumulated more consistently in the liver than in the blood plasma or muscle, likely due to its role as the central organ for fat processing and storage. Blood serum and muscle fatty acid profiles were influenced by diet, but fluctuated over-time. The low level of correlation between diet and muscle FA profiles is likely a result of low levels of fat (<1%) in the muscle and the domination of structural, cell-membrane phospholipids in shark muscle tissues. Our findings describe inter-tissue differences in the incorporation of fatty acids from the diet to consumer, which should be taken into account when interpreting dietary patterns from fatty acid profiles.


Aetiology of paediatric pneumonia with effusion in the Dominican Republic and the potential impact of pneumococcal conjugate vaccines.
Pleural effusion is a serious complication of pneumonia, and Streptococcus pneumoniae is a leading cause. We describe the aetiology of pneumonia with effusion among children in the Dominican Republic before the introduction of the 13-valent pneumococcal conjugate vaccine (PCV) in 2013 and the performance characteristics of a rapid immunochromatographic test (ICT) for detecting S. pneumoniae in pleural fluid. From July 2009 to June 2011, we enrolled children <15 years old admitted with pneumonia and pleural effusion to Robert Reid Cabral Children's Hospital, Dominican Republic. Pleural fluid was tested by culture, polymerase chain reaction (PCR) for bacterial (S. pyogenes, S. pneumoniae) and viral (respiratory syncytial virus and human rhinovirus) pathogens, and by ICT for S. pneumoniae. We calculated the performance of ICT and culture compared with PCR. Among 121 cases, the median age was 31 months (range 1 week to 14 years). Pleural fluid culture (n = 121) and PCR testing (n = 112) identified an aetiology in 85 (70.2%) cases, including 62 S. pneumoniae (51.2%) and 19 Staphylococcus aureus (15.7%). The viruses tested were not detected. The most prevalent pneumococcal serotypes were 14 (n = 20), 1 (n = 13), and 3 (n = 12). Serotype coverage of the 10- and 13-valent PCVs would be 70.5% and 95.1%, respectively. The sensitivity of point-of-care ICT was 100% (95% confidence interval [CI] 94.1%-100%), while specificity was 86.3% (95% CI 73.7%-94.3%). S. pneumoniae caused more than half of paediatric pneumonia with effusion cases; introduction of PCV in the Dominican Republic could reduce the burden by 36-49%. ICT is a practical, valid diagnostic tool for clinical care and surveillance in settings with limited laboratory capacity.


The muscle pump function in chronic venous insufficiency: plethysmographic results.
The frequency and the severity of skin lesions were studied in patients with chronic venous insufficiency compared to the function of the muscle pump measured with plethysmography. The study showed that the frequency of skin symptoms parallels the muscle pump dysfunction. Measuring the ambulatory volume change on the calf level and the refilling time required for the normalisation of the calf volume after dorsiflexions of the foot gives suitable parameters for the evaluation of the muscle pump. The plethysmography is a valuable method to quantify the grade of venous insufficiency.


Beta-adrenergic receptor blockers. Adverse effects and drug interactions.
Adverse effects of beta-adrenergic receptor blocking drugs can be divided into two categories: 1) those that result from known pharmacological consequences of beta-adrenergic receptor blockade; and 2) other reactions that do not appear to result from beta-adrenergic receptor blockade. Adverse effects of the first type include bronchospasm, heart failure, prolonged hypoglycemia, bradycardia, heart block, intermittent claudication, and Raynaud's phenomenon. Neurological reactions include depression, fatigue, and nightmares. It is not yet proven whether the beta 1-selective adrenergic blockers or those with partial agonist activity reduce the overall frequency of adverse reactions seen with propranolol. Patient age does not appear, in itself, to be associated with more beta-blocker side effects. Side effects of the second category are rare. They include an unusual oculomucocutaneous reaction and the possibility of oncogenesis. There are also many drugs that interact with beta-blockers, which may increase toxicity. Finally, there are specific patient characteristics where one beta-blocker may be more effective and safer than another.


HEI10 is subject to phase separation and mediates RPA1a degradation during meiotic interference-sensitive crossover formation.
Reciprocal exchanges of DNA between homologous chromosomes during meiosis, or crossovers (COs), shuffle genetic information in gametes and progeny. In many eukaryotes, the majority of COs (class I COs) are sensitive to a phenomenon called interference, which influences the occurrence of closely spaced double COs. Class I COs depend on a group of factors called ZMM (Zip, Msh, Mer) proteins including HEI10 (Human Enhancer of Invasion-10). However, how these proteins are recruited to class I CO sites is unclear. Here, we show that HEI10 forms foci on chromatin via a liquid-liquid phase separation (LLPS) mechanism that relies on residue Ser70. A HEI10S70F allele results in LLPS failure and a defect in class I CO formation. We further used immunoprecipitation-mass spectrometry to identify RPA1a (Replication Protein A 1) as a HEI10 interacting protein. Surprisingly, we find that RPA1a also undergoes phase separation and its ubiquitination and degradation are directly regulated by HEI10. We also show that HEI10 is required for the condensation of other class I CO factors. Thus, our results provide mechanistic insight into how meiotic class I CO formation is controlled by HEI10 coupling LLPS and ubiquitination.


Effect of repetitive firing on passive fit of metal substructure produced by the laser sintering in implant-supported fixed prosthesis.
The aim of the present study was to investigate the passive fit of metal substructure after repetitive firing processes in implant-supposed prosthesis. Five implants (4 mm diameter and 10 mm length) were placed into the resin-based mandibular model and 1-piece of screw-retained metal substructure was produced with the direct metal laser sintering (DMSL) method using Co-Cr compound (n = 10). The distance between the marked points on the multiunit supports and the marginal end of the substructure was measured using a scanning electron microscope (SEM) at each stage (metal, opaque, dentin, and glaze). 15 measurements were taken from each prosthesis, and 150 measurements from 10 samples were obtained. In total, 600 measurements were carried out at 4 stages. One-way ANOVA test was used for statistical evaluation of the data. When the obtained marginal range values were examined, differences between groups were found to be statistically significant (P<.001). The lowest values were found in the metal stage (172.4 ± 76.5 µm) and the highest values (238.03 ± 118.92 µm) were determined after glaze application. When the interval values for groups are compared with pairs, the differences between metal with dentin, metal with glaze, opaque with dentin, opaque with glaze, and dentin with glaze were found to be significant (P<.05), whereas the difference between opaque with metal was found to be insignificant (P=.992). Passive fit of 1-piece designed implant-retained fixed prosthesis that is supported by multiple implants is negatively affected by repetitive firing processes.


Indication for nateglinide in type 2 diabetes mellitus.
Nateglinide is a novel rapid- and short-acting insulin secretagogue that ameliorates postprandial hyperglycemia by improving insulin secretory dynamics to a near normal level more effectively than sulfonylureas. Recent epidemiological studies have demonstrated that postprandial hyperglycemia can result in arteriosclerosis, and that advanced arteriosclerosis is present in the initial stage of impaired glucose tolerance. Since postprandial hyperglycemia could be well treated by nateglinide, we examined the background factors of type 2 diabetic patients to determine the optimal indication for nateglinide. Our results indicate that nateglinide is most effective in young and obese patients. Furthermore, fewer responders had microangiopathy or were previously on oral hypoglycemic agents or sulfonylureas compared with non-responders. Although nateglinide is generally indicated for patients with mild HbA1c level, the present findings indicate that the drug was effective in the aforementioned patients regardless of pretreatment HbA1c levels. In one obese patient, nateglinide improved late hyperinsulinemia to near normal secretory dynamics. Our findings suggest that nateglinide is a physiologically preferable and useful drug for early type 2 diabetes without microangiopathy.


Functional Clusters of Neurons in Layer 6 of Macaque V1.
Layer 6 appears to perform a very important role in the function of macaque primary visual cortex, V1, but not enough is understood about the functional characteristics of neurons in the layer 6 population. It is unclear to what extent the population is homogeneous with respect to their visual properties or if one can identify distinct subpopulations. Here we performed a cluster analysis based on measurements of the responses of single neurons in layer 6 of primary visual cortex in male macaque monkeys (Macaca fascicularis) to achromatic grating stimuli that varied in orientation, direction of motion, spatial and temporal frequency, and contrast. The visual stimuli were presented in a stimulus window that was also varied in size. Using the responses to parametric variation in these stimulus variables, we extracted a number of tuning response measures and used them in the cluster analysis. Six main clusters emerged along with some smaller clusters. Additionally, we asked whether parameter distributions from each of the clusters were statistically different. There were clear separations of parameters between some of the clusters, particularly for f1/f0 ratio, direction selectivity, and temporal frequency bandwidth, but other dimensions also showed differences between clusters. Our data suggest that in layer 6 there are multiple parallel circuits that provide information about different aspects of the visual stimulus.SIGNIFICANCE STATEMENT The cortex is multilayered and is involved in many high-level computations. In the current study, we have asked whether there are subpopulations of neurons, clusters, in layer 6 of cortex with different functional tuning properties that provide information about different aspects of the visual image. We identified six major functional clusters within layer 6. These findings show that there is much more complexity to the circuits in cortex than previously demonstrated and open up a new avenue for experimental investigation within layers of other cortical areas and for the elaboration of models of circuit function that incorporate many parallel pathways with different functional roles.


Successful medical management of pseudomembranous cystitis in three cats with lower urinary tract obstruction.
The present case series describes the clinical course and outcome of three cats diagnosed with pseudomembranous cystitis. This is an uncommon presentation of lower urinary tract obstruction but can be easily be identified by ultrasonography, revealing severe bladder wall thickening and thin hyperechoic luminal strips. The condition can be secondary to severe bacterial urinary tract infection. All cats were successfully treated with medical management only, mainly based on antimicrobials and individualised supportive therapy. Further evaluation of this condition is necessary in order to determine potential underlying aetiologies, pathophysiological mechanisms and the most appropriate standardised treatment.


Expression of bonnet monkey (Macaca radiata) zona pellucida-3 (ZP3) in a prokaryotic system and its immunogenicity.
An internal fragment (978 bp) corresponding to the bonnet monkey (Macaca radiata) ZP3, excluding the N-terminus signal sequence and the C-terminus transmembrane-like domain, was amplified by PCR from a full-length cDNA clone. The amplified Bam HI and SacI restricted fragment was cloned in frame downstream of the T5 promoter under lac operator control for expression in the pQE-30 vector. Recombinant ZP3 (r-ZP3) was expressed as a poly-histidine fusion protein in E. coli strains SG13009[pREP4] and BL-21(DE3). Immunoblot with a murine monoclonal antibody, MA-451 (raised against porcine ZP3 beta-a homologue of bonnet ZP3, and cross-reactive with bonnet zona pellucida) revealed a predominant band of 50 kDa besides degraded fragments. Optimum expression of r-ZP3 was observed at 0.5 mM IPTG. Antisera generated in monkeys against synthetic peptides from the N-(23-45 aa residues) and C-(300-322 and 324-347 aa residues) termini of the deduced bonnet monkey precursor ZP3 sequence reacted with the r-ZP3 protein in ELISA. The r-ZP3 expressed in SG13009[pREP4] was purified on Ni-NTA resin under denaturing conditions and conjugated with diphtheria toxoid (DT). Immunization of a female rabbit and six female bonnet monkeys with the r-ZP3-DT conjugate generated antibodies reactive with r-ZP3 in ELISA. Rabbit r-ZP3 antiserum reacted with porcine ZP3 beta and bonnet r-ZP3 but failed to react with porcine ZP3 alpha in a Western blot. Moreover, antisera when tested by indirect immunofluorescence on bonnet monkey ovarian sections, showed positive fluorescence with zona pellucida. The availability of r-ZP3 will further help in evaluating its efficacy for fertility regulation and understanding the autoimmune oophoritis associated with ZP3 immunization in nonhuman primates.


Perceived Effectiveness and Motivations for the Use of Web-Based Mental Health Programs: Qualitative Study.
The prevalence of depression is high and has been stable despite increased treatment, research, and dissemination. People encounter barriers to seeking traditional mental health services, which could be mitigated by using web-based prevention methods. This study aims to understand what people at high risk for depression perceive as effective aspects of web-based mental health programs and what motivates people at high risk for depression to use web-based mental health programs. We conducted an inductive content analysis using telephone interview data from 77 participants at high risk for depression who were recruited from 2 randomized controlled trials (RCTs). Participants from the first RCT were working men who had been randomly assigned to 1 of the following 3 groups: control group, who had access to general depression information from a website called BroMatters; intervention group 1, who had access to the BroMatters website along with the associated BroHealth web-based mental health program; and intervention group 2, who had access to the BroMatters website, the BroHealth web-based mental health program, and telephone sessions with a life coach. Participants from the second RCT were men and women who had been assigned to the intervention group, who received access to the HardHat web-based mental health program, or the control group, who only received access to the HardHat web-based mental health program following completion of the RCT. Participants for this inductive content analysis study were recruited from the intervention groups in both RCTs. Two groups of participants (n=41 and n=20) were recruited from the BroHealth RCT, and a third group comprised 16 participants that were recruited from the HardHat RCT. We generated four categories regarding the perceived effectiveness of web-based programs and five categories related to what motivates the use of web-based programs. Participants identified awareness, program medium and functionality, program content, and coaches as categories related to the effectiveness of the programs. Categories of motivators to use web-based programs included providing reminders or incentives, promotion of the programs, providing appropriate medium and functionality, appropriate content, and perceived need. The final category related to motivators reflects perceptions of participants who were either unsure about what motivates them or believed that there is no way to motivate use. Conflicting evidence was obtained regarding the perceived effectiveness of aspects of the content and functionality of web-based programs. In general, web-based mental health programs were perceived to help increase mental health awareness, especially when it includes live access to a coach. However, the results also revealed that it is difficult to motivate people to begin using web-based mental health programs. Strategies that may motivate the use of such programs include perceived personal need, effective promotion, providing incentives and reminders, and improving functionality.


Cleft palate and craniofacial teams in the United States and Canada: a national survey of team organization and standards of care. The American Cleft Palate-Craniofacial Association (ACPA) Team Standards Committee.
This study is the first comprehensive national survey of the organization, function, and composition of cleft palate and craniofacial teams in the U.S. and Canada. Complete descriptions of cleft and craniofacial teams are not currently provided in the literature, and this study will provide an overview for health services research and policy use. Conducted by a national organization, this study examines teams in detail using a pretested and standardized methodology. All known (n = 296) North American cleft palate and craniofacial teams were contacted for team listing purposes using a self-assessment method developed by an interdisciplinary committee of national stature. Team clinical leaders classified their teams into several possible categories and provided data on team care. The response rate was 83.4% (n = 247). The distribution of listed teams was: 105 (42.5%) cleft palate teams, 102 (41.3%) craniofacial teams (including craniofacial teams that are both cleft palate and craniofacial teams), 12 (4.9%) geographically listed teams, and 28 (11.3%) other teams (including interim cleft palate teams, low-density cleft palate teams, and evaluation and treatment review cleft palate teams). Eighty-five percent of all teams systematically collected and stored clinical data on their team's patient population in the past year. Furthermore, 50% of all teams had a quality assurance program in place to measure treatment outcomes. Other findings presented include the annual number of face-to-face team meetings; new and follow-up patient censuses; and surgical rates for initial repair of cleft lip/palate, orthognathic/osteotomy procedures, and intracranial/craniofacial procedures. Two of five North American teams classify themselves as having the capacity to provide both cleft palate and craniofacial care. An additional two of five teams limit their primary role to cleft palate care. Issues are raised regarding the distribution of teams, the regionalization of craniofacial services, health policy, and resource allocation.


[Advances in revision surgery after primary total hip arthroplasty for Crowe type Ⅳ developmental dysplasia of the hip].
To review research advances of revision surgery after primary total hip arthroplasty (THA) for patients with Crowe type Ⅳ developmental dysplasia of the hip (DDH). The recent literature on revision surgery after primary THA in patients with Crowe type Ⅳ DDH was reviewed. The reasons for revision surgery were analyzed and the difficulties of revision surgery, the management methods, and the related prosthesis choices were summarized. Patients with Crowe type Ⅳ DDH have small anteroposterior diameter of the acetabulum, large variation in acetabular and femoral anteversion angles, severe soft tissue contractures, which make both THA and revision surgery more difficult. There are many reasons for patients undergoing revision surgery after primary THA, mainly due to aseptic loosening of the prosthesis. Therefore, it is necessary to restore anatomical structures in primary THA, as much as possible and reduce the generation of wear particles to avoid postoperative loosening of the prosthesis. Due to the anatomical characteristics of Crowe type Ⅳ DDH, the patients have acetabular and femoral bone defects, and the repair and reconstruction of bone defects become the key to revision surgery. The acetabular side is usually reconstructed with the appropriate acetabular cup or combined metal block, Cage, or custom component depending on the extent of the bone defect, while the femoral side is preferred to the S-ROM prosthesis. In addition, the prosthetic interface should be ceramic-ceramic or ceramic-highly cross-linked polyethylene wherever possible. The reasons leading to revision surgery after primary THA in patients with Crowe type Ⅳ DDH and the surgical difficulties have been clarified, and a large number of clinical studies have proposed corresponding revision modalities based on which good early- and mid-term outcomes have been obtained, but further follow-up is needed to clarify the long-term outcomes. With technological advances and the development of new materials, personalized prostheses for these patients are expected to become a reality.


The importance of alpha-fodrin antibodies in the diagnosis of Sjögren's syndrome.
We wanted to determine the prevalence of IgA and IgG antibodies against alpha-fodrin in the patients with primary and secondary Sjögren's syndrome (SS) and also to compare with anti-Ro and anti-La antibodies in the diagnosis of SS. We tested the prevalence of anti-alpha-fodrin IgA, IgG, anti-Ro, anti-La antibodies, anti-nuclear antibodies (ANA) and rheumatoid factor (RF) in naive patients with primary (n = 20) and secondary SS (n = 20) (Rheumatoid Arthritis [RA]+SS, n = 10; Systemic Lupus Erythematosus [SLE] + SS, n = 10), RA (n = 10), SLE (n = 10) and in healthy controls (n = 20). Salivary gland biopsies were performed in the patients with primary and secondary SS. In primary SS, anti-alpha-fodrin IgA, IgG, anti-Ro and anti-La antibodies were detected as 20, 10, 55 and 20% respectively. In RA + SS, anti-alpha-fodrin IgA was detected to be 10% and IgG was negative; however, anti-Ro antibodies and anti-La antibodies were found to be 40% and 20% respectively. In SLE + SS, anti-alpha-fodrin IgA was found to be 20% and IgG was found to be 10%, but anti-Ro and anti-La antibodies were found to be 90% and 20% respectively. Alpha-fodrin antibodies were not detected in RA, SLE and healthy controls. The detection of anti-alpha-fodrin antibodies by used ELISA does not give much contribution to the diagnosis of SS, and anti-Ro and anti-La are still useful serological markers in the diagnosis of SS.


Detergent- and phospholipid-based reconstitution systems have differential effects on constitutive activity of G-protein-coupled receptors.
A hallmark of G-protein-coupled receptors (GPCRs) is the conversion of external stimuli into specific cellular responses. In this tightly-regulated process, extracellular ligand binding by GPCRs promotes specific conformational changes within the seven transmembrane helices, leading to the coupling and activation of intracellular "transducer" proteins, such as heterotrimeric G proteins. Much of our understanding of the molecular mechanisms that govern GPCR activation is derived from experiments with purified receptors reconstituted in detergent micelles. To elucidate the influence of the phospholipid bilayer on GPCR activation, here we interrogated the functional, pharmacological, and biophysical properties of a GPCR, the β2-adrenergic receptor (β2AR), in high-density lipoprotein (HDL) particles. Compared with detergent-reconstituted β2AR, the β2AR in HDL particles had greatly enhanced levels of basal (constitutive) activity and displayed increased sensitivity to agonist activation, as assessed by activation of heterotrimeric G protein and allosteric coupling between the ligand-binding and transducer-binding pockets. Using 19F NMR spectroscopy, we directly linked these functional differences in detergent- and HDL-reconstituted β2AR to a change in the equilibrium between inactive and active receptor states. The contrast between the low levels of β2AR constitutive activity in cells and the high constitutive activity observed in an isolated phospholipid bilayer indicates that β2AR basal activity depends on the reconstitution system and further suggests that various cellular mechanisms suppress β2AR basal activity physiologically. Our findings provide critical additional insights into GPCR activation and reveal how dramatically reconstitution systems can impact membrane protein function.


Visual effects of lesions of cortical area V2 in macaques.
Ibotenic acid lesions were placed in two monkeys in a portion of cortical area V2 that corresponds to a lower quadrant of the visual field extending approximately 3-7 degrees from the fovea. For purposes of comparison, another lesion was placed in area V1 in one animal. A wide range of visual capacities were then measured, using a discrimination between vertical and horizontal orientation, in and near the affected regions of the visual field. Visual acuity declined sharply as the test stimulus approached the visual field location corresponding to the V1 lesion, and no threshold could be measured at its center. In contrast, lesions of area V2 caused no measurable decrease in acuity, nor was there any substantial effect on several measures of contrast sensitivity. Subsequently, two types of more complex visual discriminations were measured (also using a vertical-horizontal discrimination), and these discriminations were severely disrupted by V2 lesions. The first discrimination was of the orientation of two parallel lines of five colinear dots each. We measured the number of background dots that would bring the discrimination to threshold, and this number of dots was greatly decreased by a V2 lesion. The second discrimination was of the orientation of a group of three distinctive texture elements embedded in a six by six element texture. This task could not be done in the visual field region affected by the V2 lesion when the distinctive elements differed in orientation from the others. Control experiments showed that the discrimination could be done when the three distinctive elements differed in size or color. These results suggest that cortical area V2 is not needed for some low-level discriminations, but may be essential for tasks involving complex spatial discriminations.


Spiritual Fitness for Military Veterans: A Curriculum Review and Impact Evaluation Using the Duke Religion Index (DUREL).
Suicide rates among military veterans exceed those found in the general population. While the exact reasons for these high rates are unknown, contributing factors may include the military's perceived rejection of patient identities, creating barriers to mental health care within the clinical sector and a mandate for prevention programs. Spiritual fitness has emerged over the last decade as an important concept in human performance optimization and is included among holistic approaches to developing and maintaining mentally fit fighting forces. In attempts to better understand the role that spiritual fitness and religion play in mitigating and/or reducing suicide risk among veterans, the aims of this study were twofold (1) to assess the utility of the Duke Religion Index as a psychometric instrument for use with veterans completing spiritual fitness training and (2) to offer a post-intervention process evaluation of the spiritual fitness module from one resilience program offered to military veterans of Iraq and Afghanistan in 2016. Twenty-eight attendees at the JRWI Wellness Resilient Leadership Retreat completed post-retreat surveys to assess their satisfaction with the coursework and specifically, to assess the spiritual fitness module of the resiliency retreat's curriculum. In total, the research team reviewed 25 completed post-intervention survey responses (89.3% response rate). Descriptive statistics indicated that respondents (n = 25) were subjectively religious, defined as belief in a higher power practiced in ritualized ways. Over half of program participants indicated they (a) attended religious meetings at least once a week and (b) engaged in private religious activity-such as meditation-at least once a day. Results showed that most program participants reported that the spiritual fitness skills learned during the resilient leadership program were useful (88%) (Z = 3.000, p < 0.001). Additionally, most program participants reported their interest in spiritual exploration was affirmed, renewed, or raised as a result of attending the peer-led resilient leadership program (76%) (Z = 6.000, p = 0.015). Culturally informed prevention programs that emphasize spiritual fitness are indicated for use in veteran outreach and well-being programs. More detailed research is needed to assess curriculum specifics.


Epidemiological history and genomic characterization of non-D1 HBV strains identified in Iran.
Hepatitis B virus (HBV) has been classified into eight genotypes and forty subgenotypes. Genotype D of HBV is the most worldwide distributed genotype and HBV subgenotype D1 has been isolated from Iranian patients. To characterize for the first time complete genomes of recently emerged non-D1 strains in Iran. HBV complete genomes isolated from 9 Iranian HBV carriers were sequenced. Different diversities of the ORFs were mapped and evolutionary history relationships were investigated. Phylogenetic analysis identified four D2 subgenotypes and five D3 subgenotypes of HBV in the studied patients. Of note, D2 strains clustered with strains from Lebanon and Syria. The time of the most recent common ancestor (TMRCA) of the first cluster of D2 was dated at 1953 (BCI=1926, 1976) while the second cluster was dated at 1947 (BCI=1911, 1978). All five Iranian D3 strains formed a monophyletic cluster with Indian strain and dated back to 1967 (BCI=1946, 1987). Surprisingly, two D3 strains had an adw2 subtype. Interestingly, more than 80% of the present strains showed precore mutations, while two isolates carried basal core promoter variation. Iranian D2 and D3 isolates were introduced on at least two and one occasion in Iran and diverged from west and south Asian HBV strains, respectively. Considering the impact of the different (sub) genotypes on clinical outcome, exploring the distinct mutational patterns of Iranian D1 and non-D1 strains is of clinical importance.


Assembly-dependent maturation cleavage in provirions of a small icosahedral insect ribovirus.
Extracts from nodavirus-infected Drosophila cells contained detergent-labile 140S "young" particles much richer than mature virions in their content of protein alpha, a precursor of coat proteins beta and gamma. Incorporation studies in infected cells showed that most newly synthesized alpha protein was assembled into young particles within a few minutes. Incubation of the particles, either in cytoplasmic extracts or after purification, resulted in spontaneous first-order cleavage of alpha protein to form beta-plus-gamma chains. Alpha protein that was not associated with particles failed to cleave. Cleavage was accompanied by a marked increase in detergent stability of the particles and was unaffected by a broad spectrum of protease inhibitors or by coating with precipitating antibody. We conclude (i) that alpha chains are cleaved only after assembly into provirions, (ii) that cleavage occurs internally and is likely therefore autocatalytic, and (iii) that cleavage stabilizes the mature virus particles.


EP-cadherin in muscles and epithelia of Xenopus laevis embryos.
EP-cadherin is a novel Xenopus Ca+2-dependent adhesion molecule, which shares comparable homology with mouse E- and P-cadherins (Ginsberg, De Simone and Geiger; 1991, Development 111, 315-325). We report here the patterns of expression of this molecule in Xenopus laevis embryos at different developmental stages ranging from cleavage to postmetamorphic. EP-cadherin is already expressed in the oocyte and egg and can then be detected in close association with the membrane of all blastomeres up to late blastula stages. Starting at late gastrula stages, the level of EP-cadherin expression increases sharply in non-neural ectodermal cells, in the somites and in the notochord; it persists in endodermal cells and decreases rapidly in all migratory cells. During neurulation the level of EP-cadherin expression declines gradually in the nervous system and is undetectable here throughout later development except in the optic nerve and in the neural part of the olfactory organ. This pattern continues during later development so that in the tailbud stage and up to metamorphosis the most prominent staining is detected in the epidermis and skeletal muscle. After metamorphosis, the molecule gradually disappears from the muscle tissue and the major site of expression remains the skin. EP-cadherin is invariably present in close association with the cell membrane. In the muscle it is associated with the sarcolemma at regions of myoblast-myoblast or myotube-myotube contact. In epidermal cells, EP-cadherin is usually coexpressed with E-cadherin. Yet, while E-cadherin staining is always restricted to the basolateral aspects of the cells, EP-cadherin is often distributed throughout the plasmalemma including the apical surface.


Association between Poor Sleep Quality and Subsequent Peptic Ulcer Recurrence in Older Patients with Mild Cognitive Impairment: Examining the Role of Social Engagement.
This study aims to examine whether perceived poor sleep quality predicts subsequent recurrence of peptic ulcer disease (PUD) in older patients with mild cognitive impairment following Helicobacter pylori eradication and to investigate whether social engagement status alters this association. Of 1,689 older patients with H. pylori-infected PUD recruited from eight Grade-A hospitals in the People's Republic of China between 2011 and 2014, H. pylori was eradicated and PUD cleared in 1,538 patients by the end of 2014; 1,420 of these were followed for up to 36 months. The Kaplan-Meier method was used to compare the proportion of PUD recurrence, as confirmed with esophagogastroduodenoscopy, among older patients with different levels of sleep quality and social engagement statuses. Multivariate Cox-proportional hazards models were performed to examine the association between sleep quality and PUD recurrence, and the role of social engagement in altering this relationship. The results showed that PUD recurrence was more prevalent in poor (10.8%) compared with good sleepers (5.5%). However, increased and continued social engagement reduced the proportion to 7.2% and 8.2% among poor sleepers, respectively. Poor sleep quality was associated with subsequent PUD recurrence (hazard ratio [HR] 1.965 (1.002, 3.518)). However, no significant difference was observed between good and poor sleepers who reported increased (HR 1.428 (0.736, 2.380)) and continued (HR 1.273 (0.915, 2.492)) social engagement, suggesting that increased and continued social engagement prevented the effect of poor sleep quality on PUD recurrence. To conclude, poor sleep quality is associated with subsequent PUD recurrence. However, increased and continued social engagement may moderate this association.


The efficacy of topical ciprofloxacin and norfloxacin in the treatment of experimental Pseudomonas keratitis.
An aminoglycoside-resistant strain of Pseudomonas aeruginosa was injected intrastromally into the corneas of rabbits, and keratitis was allowed to develop over a 22-h period. Rabbits were treated with either 0.75% ciprofloxacin, 1% norfloxacin, or 1.36% tobramycin administered topically every 15 min for 1 h and then every 30 min for the following 3 h. All therapy ceased 26 h postinoculation. Rabbits were killed 1 h after the treatment, and the number of bacteria per cornea were quantified in terms of bacterial colony-forming units. Aqueous humor specimens were obtained from rabbits receiving norfloxacin and ciprofloxacin, and bioassays were performed to determine drug concentration. Ciprofloxacin caused a 5 log reduction in the number of bacterial colony-forming units, as compared with untreated controls (p less than 0.0001); it also produced a significantly greater reduction in bacterial colony-forming units than either norfloxacin or fortified tobramycin drops (p less than 0.0001). Norfloxacin produced a 2 log reduction in bacterial colony-forming units, as compared with untreated controls (p less than 0.0001). The mean aqueous concentration of norfloxacin (7.5 micrograms/ml) was substantially less than that achieved by ciprofloxacin (30.5 micrograms/ml). We conclude that ciprofloxacin may be a useful broad spectrum, topical chemotherapeutic agent in the therapy of aminoglycoside-resistant P. aeruginosa keratitis.


ETV2 expression marks blood and endothelium precursors, including hemogenic endothelium, at the onset of blood development.
ETV2 has been identified as an important player in embryonic hematopoiesis. However, the cell populations in which this transcription factor is expressed and operates during blood specification remain to be fully characterized. Here we address these issues using ES cells and a transgenic mouse line expressing green fluorescent protein (GFP) under the control of ETV2 regulatory elements, allowing us to observe the tight association between ETV2 expression and the initiation of hematopoiesis. Both in differentiating ES cells and gastrulating embryos ETV2::GFP is mostly found co-expressed with endothelial markers and defines a subset of cells with greatly enriched primitive erythroid potential. Upon culture ETV2::GFP cells rapidly up-regulate CD41, down-regulate endothelium cell surface markers and generate definitive hematopoietic progenitors. Altogether these characteristics represent the hallmark of hemogenic endothelium cells, a specialized endothelium originating from the hemangioblast and giving rise to hematopoietic cells. Importantly, ETV2 deficiency results in a complete absence of hemogenic endothelium in differentiating ES cells and gastrulating embryos. Altogether our data reveal that ETV2 marks hemogenic endothelium in gastrulating embryos and is absolutely required for the formation of this precursor at the onset of hematopoiesis. These results enhance our understanding of embryonic hematopoiesis and the factors driving hemogenic endothelium specification.


Synthesis and biological evaluation of Ribo 7-N/O/S pyrimidine 9-deaza C-nucleoside analogs as new antiviral agents for inhibiting HCV RNA-dependent RNA polymerases.
Hepatitis C infection is caused by the bloodborne pathogen hepatitis C virus (HCV) and can lead to serious liver diseases and, ultimately, death if the treatment is ineffective. This work reports the synthesis and preclinical evaluation of 7 novel 9-O/N/S pyrimidine nucleosides, including compound 12, the triphosphate of known compound 7b. The nucleosides are 9-deaza modifications of adenosine and guanosine with β-2'-C-methyl substituent on the ribose. Within this series of compounds, a 9-deaza furopyrimidine analog of adenosine, compound 7b, showed high anti-HCV activity in vitro, good stability, low toxicity, and low genotoxicity when administrated in low doses, and an adequate pharmacokinetics profile. An improved synthesis of compound 7b compared to a previous study is also reported. Compound 12 was synthesized as a control to verify phosphorylation of 7b occurred in vivo.


Clinical features differentiating biliary atresia from other causes of neonatal cholestasis.
This study determined any clinical features which may help to differentiate biliary atresia (BA) from other causes of neonatal cholestasis (NC). A prospective and observational study was conducted on consecutive infants with NC referred to the University of Malaya Medical Centre, Malaysia, between November 1996 and May 2004. The 3 most common causes of cholestasis among the 146 infants with NC studied were idiopathic neonatal hepatitis (n = 63, 43%), BA (n = 35, 24%) and congenital cytomegalovirus hepatitis (n = 13, 9%). Common clinical features at presentation were jaundice (100%), hepatomegaly (95%), splenomegaly (52%) and pale stools (47%). Three clinical features noted to be sensitive for BA were the presence of acholic or variably acholic stools on admission, a liver which was firm/hard in consistency and a palpable liver of ≥4 cm (sensitivity of 77%, 80% and 94%, respectively), but the corresponding specificity was poor (51%, 65% and 39%, respectively). The stools of 2 children with BA were pigmented initially but became acholic subsequently. We did not find any single clinical feature with sufficient sensitivity and specificity to differentiate BA from other causes of NC. Repeated inspection of stools colour is necessary as occasionally, patients with BA may have initial pigmented stools. Biochemical assessment and imaging studies are important in the assessment of any infant with NC.


Myocardial protection by insulin at reperfusion requires early administration and is mediated via Akt and p70s6 kinase cell-survival signaling.
The "metabolic cocktail" comprising glucose-insulin-potassium administrated at reperfusion reduces infarct size in the in vivo rat heart. We propose that insulin is the major component mediating this protection and acts via Akt prosurvival signaling. This hypothesis was studied in isolated perfused rat hearts (measuring infarct size to area of risk [%]) subjected to 35 minutes regional myocardial ischemia and 2 hours reperfusion. Insulin administered at the onset of reperfusion attenuated infarct size by >/=45% versus control hearts (P<0.001). Insulin-mediated cardioprotection was found to be independent of the presence of glucose at reperfusion. Moreover, the cell survival benefit of insulin is temporally dependent, in that insulin administration from the onset of reperfusion and maintained for either 15 minutes or for the duration of reperfusion reduced infarct size. In contrast, protection was abrogated if insulin administration was delayed until 15 minutes into reperfusion. Pharmacological inhibition of both upstream and downstream signals in the Akt prosurvival pathway abolished the cardioprotective effects of insulin. Here coadministration of insulin with the tyrosine kinase inhibitor lavendustin A, the phosphatidylinositol3-kinase (PI3-kinase) inhibitor wortmannin, and mTOR/p70s6 kinase inhibitor rapamycin abolished cardioprotection. Steady-state levels of activated/phosphorylated Akt correlated with insulin administration. Finally, downstream prosurvival targets of Akt including p70s6 kinase and BAD were modulated by insulin. In conclusion, insulin administration at reperfusion reduces myocardial infarction, is dependent on early administration during reperfusion, and is mediated via Akt and p70s6 kinase dependent signaling pathway. Moreover, BAD is maintained in its inert phosphorylated state in response to insulin therapy.


The 5-HT3B subunit is a major determinant of serotonin-receptor function.
The neurotransmitter serotonin (5-hydroxytryptamine or 5-HT) mediates rapid excitatory responses through ligand-gated channels (5-HT3 receptors). Recombinant expression of the only identified receptor subunit (5-HT3A) yields functional 5-HT3 receptors. However, the conductance of these homomeric receptors (sub-picosiemens) is too small to be resolved directly, and contrasts with a robust channel conductance displayed by neuronal 5-HT3 receptors (9-17 pS). Neuronal 5-HT3 receptors also display a permeability to calcium ions and a current-voltage relationship that differ from those of homomeric receptors. Here we describe a new class of 5-HT3-receptor subunit (5-HT3B). Transcripts of this subunit are co-expressed with the 5-HT3A subunit in the amygdala, caudate and hippocampus. Heteromeric assemblies of 5-HT3A and 5-HT3B subunits display a large single-channel conductance (16 pS), low permeability to calcium ions, and a current-voltage relationship which resembles that of characterized neuronal 5-HT3 channels. The heteromeric receptors also display distinctive pharmacological properties. Surprisingly, the M2 region of the 5-HT3B subunit lacks any of the structural features that are known to promote the conductance of related receptors. In addition to providing a new target for therapeutic agents, the 5-HT3B subunit will be a valuable resource for defining the molecular mechanisms of ion-channel function.


Exercise augments the acute anabolic effects of intradialytic parenteral nutrition in chronic hemodialysis patients.
Decreased dietary protein intake and hemodialysis (HD)-associated protein catabolism are among several factors that predispose chronic hemodialysis (CHD) patients to uremic malnutrition and associated muscle wasting. Intradialytic parenteral nutrition (IDPN) acutely reverses the net negative whole body and forearm muscle protein balances observed during the HD procedure. Exercise has been shown to improve muscle protein homeostasis, especially if performed with adequately available intramuscular amino acids. We hypothesized that exercise performance would provide additive anabolic effects to the beneficial effects of IDPN. We studied six CHD patients at two separate HD sessions: 1) IDPN administration only and 2) IDPN + exercise. Patients were studied 2 h before, during, and 2 h after an HD session by use of a primed constant infusion of l-[1-(13)C]leucine and l-[ring-(2)H(5)] phenylalanine. Exercise combined with IDPN promoted additive twofold increases in forearm muscle essential amino acid uptake (455 +/- 105 vs. 229 +/- 38 nmol.100 ml(-1).min(-1), P < 0.05) and net muscle protein accretion (125 +/- 37 vs. 56 +/- 30 microg.100 ml(-1).min(-1), P < 0.05) during HD compared with IDPN alone. Measurements of whole body protein homeostasis and energy expenditure were not altered by exercise treatment. In conclusion, exercise in the presence of adequate nutritional supplementation has potential as a therapeutic intervention to blunt the loss of muscle mass in CHD patients.


Quantitative PCR primer design affects quantification of dsRNA-mediated gene knockdown.
RNA interference (RNAi) is a powerful tool for studying functions of candidate genes in both model and nonmodel organisms and a promising technique for therapeutic applications. Successful application of this technique relies on the accuracy and reliability of methods used to quantify gene knockdown. With the limitation in the availability of antibodies for detecting proteins, quantitative PCR (qPCR) remains the preferred method for quantifying target gene knockdown after dsRNA treatment. We evaluated how qPCR primer binding site and target gene expression levels affect quantification of intact mRNA transcripts following dsRNA-mediated RNAi. The use of primer pairs targeting the mRNA sequence within the dsRNA target region failed to reveal a significant decrease in target mRNA transcripts for genes with low expression levels, but not for a highly expressed gene. By contrast, significant knockdown was detected in all cases with primer pairs targeting the mRNA sequence extending beyond the dsRNA target region, regardless of the expression levels of the target gene. Our results suggest that at least for genes with low expression levels, quantifying the efficiency of dsRNA-mediated RNAi with primers amplifying sequences completely contained in the dsRNA target region should be avoided due to the risk of false-negative results. Instead, primer pairs extending beyond the dsRNA target region of the mRNA transcript sequences should be used for accurate and reliable quantification of silencing efficiency.


Piezoelectric Versus Conventional Rotary Techniques for Impacted Third Molar Extraction: A Meta-analysis of Randomized Controlled Trials.
Impacted third molars are frequently encountered in clinical work. Surgical removal of impacted third molars is often required to prevent clinical symptoms. Traditional rotary cutting instruments are potentially injurious, and piezosurgery, as a new osteotomy technique, has been introduced in oral and maxillofacial surgery. No consistent conclusion has been reached regarding whether this new technique is associated with fewer or less severe postoperative sequelae after third molar extraction.The aim of this study was to compare piezosurgery with rotary osteotomy techniques, with regard to surgery time and the severity of postoperative sequelae, including pain, swelling, and trismus.We conducted a systematic literature search in the Cochrane Library, PubMed, Embase, and Google Scholar.The eligibility criteria of this study included the following: the patients were clearly diagnosed as having impacted mandibular third molars; the patients underwent piezosurgery osteotomy, and in the control group rotary osteotomy techniques, for removing impacted third molars; the outcomes of interest include surgery time, trismus, swelling or pain; the studies are randomized controlled trials.We used random-effects models to calculate the difference in the outcomes, and the corresponding 95% confidence interval. We calculated the weighted mean difference if the trials used the same measurement, and a standardized mean difference if otherwise.A total of seven studies met the eligibility criteria and were included in our analysis. Compared with rotary osteotomy, patients undergoing piezosurgery experienced longer surgery time (mean difference 4.13 minutes, 95% confidence interval 2.75-5.52, P < 0.0001). Patients receiving the piezoelectric technique had less swelling at postoperative days 1, 3, 5, and 7 (all Ps ≤0.023). Additionally, there was a trend of less postoperative pain and trismus in the piezosurgery groups.The number of included randomized controlled trials and the sample size of each trial were relatively small, double blinding was not possible, and cost analysis was unavailable due to a lack of data.Our meta-analysis indicates that although patients undergoing piezosurgery experienced longer surgery time, they had less postoperative swelling, indicating that piezosurgery is a promising alternative technique for extraction of impacted third molars.


Swarm intelligence for mixed-variable design optimization.
Many engineering optimization problems frequently encounter continuous variables and discrete variables which adds considerably to the solution complexity. Very few of the existing methods can yield a globally optimal solution when the objective functions are non-convex and non-differentiable. This paper presents a hybrid swarm intelligence approach (HSIA) for solving these nonlinear optimization problems which contain integer, discrete, zero-one and continuous variables. HSIA provides an improvement in global search reliability in a mixed-variable space and converges steadily to a good solution. An approach to handle various kinds of variables and constraints is discussed. Comparison testing of several examples of mixed-variable optimization problems in the literature showed that the proposed approach is superior to current methods for finding the best solution, in terms of both solution quality and algorithm robustness.


Revision Rhinoplasty Finesse with Digital Osteotomies.
Revision rhinoplasty is a finesse procedure requiring careful consideration of all of its requisite steps. Lateral osteotomies are required in many cases and, in the setting of prior lateral osteotomies, can often be recreated with digital pressure alone, without the use of an osteotome. The advantages of doing so include decreased dead-space creation and preservation of attachments between the skeleton and overlying soft tissue.


Patient positioning method based on binary image correlation between two edge images for proton-beam radiation therapy.
A new technique based on normalized binary image correlation between two edge images has been proposed for positioning proton-beam radiotherapy patients. A Canny edge detector was used to extract two edge images from a reference x-ray image and a test x-ray image of a patient before positioning. While translating and rotating the edged test image, the absolute value of the normalized binary image correlation between the two edge images is iteratively maximized. Each time before rotation, dilation is applied to the edged test image to avoid a steep reduction of the image correlation. To evaluate robustness of the proposed method, a simulation has been carried out using 240 simulated edged head front-view images extracted from a reference image by varying parameters of the Canny algorithm with a given range of rotation angles and translation amounts in x and y directions. It was shown that resulting registration errors have an accuracy of one pixel in x and y directions and zero degrees in rotation, even when the number of edge pixels significantly differs between the edged reference image and the edged simulation image. Subsequently, positioning experiments using several sets of head, lung, and hip data have been performed. We have observed that the differences of translation and rotation between manual positioning and the proposed method were within one pixel in translation and one degree in rotation. From the results of the validation study, it can be concluded that a significant reduction in workload for the physicians and technicians can be achieved with this method.


Surface Textures of Sand Grains--an Application of Electron Microscopy: Glaciation.
To simulate glacial conditions, a mixture of ice and quartz grains was subjected to grinding under hydrostatic pressure of 500 bars. Electron microphotographs of these "synthetic" glaciated surfaces were then compared with electron microphotographs of grain surfaces which had been abraded and crushed by glacial action, and the correspondence was found to be good. Thus it appears possible to distinguish glacial sands from beach and eolian sands byexamining the surface textures of these grains with the electron microscope.


Spatial control of Cdc42 signalling by a GM130-RasGRF complex regulates polarity and tumorigenesis.
The small GTPase Cdc42 is a key regulator of polarity, but little is known in mammals about its spatial regulation and the relevance of spatial Cdc42 pools for polarity. Here we report the identification of a GM130-RasGRF complex as a regulator of Cdc42 at the Golgi. Silencing GM130 results in RasGRF-dependent inhibition of the Golgi pool of Cdc42, but does not affect Cdc42 at the cell surface. Furthermore, active Cdc42 at the Golgi is important to sustain asymmetric front-rear Cdc42-GTP distribution in directionally migrating cells. Concurrent to Cdc42 inhibition, silencing GM130 also results in RasGRF-dependent Ras-ERK pathway activation. Moreover, depletion of GM130 is sufficient to induce E-cadherin downregulation, indicative of a loss in cell polarity and epithelial identity. Accordingly, GM130 expression is frequently lost in colorectal and breast cancer patients. These findings establish a previously unrecognized role for a GM130-RasGRF-Cdc42 connection in regulating polarity and tumorigenesis.


[Computer program for the analysis of clinical diagnostic errors based on a pathology database].
The program is tested on the material of 882 autopsy protocols. All parts of clinical and pathology diagnoses were included into the generation of the data base. Automatized analysis allowed to assess main characteristics of the patient material with a histogram demonstration of group statistics according to all positions considered as well as to obtain information on the influence of more complex pathology diagnosis of a main disease, sex and age on the frequency of clinical diagnosis mistakes.


Effects of piribedil and of d-amphetamine on locomotor activity in immature rats.
The effects of piribedil, a direct dopamine (DA) agonist, and of d-amphetamine, an indirect DA agonist, on locomotor activity in 28-day old rats have been investigated. d-Amphetamine (1,2 and 4 mg/kg) increased activity in a dose dependent manner whereas only a 40 mg/kg dose of piribedil significantly increased activity; low doses of piribedil (1.25-10.0 mg/kg) had no effect on activity. Since hyperactivity induced by a direct DA agonist, such as piribedil, is thought to reflect post-synaptic DA receptor activation, the results appear to indicate that post-synaptic DA receptor mechanisms are fully developed in 28-day old rats. In contrast, as low doses of piribedil which cause sedation in adult animals failed to decrease locomotor activity under the present experimental conditions, it seems that DA autoreceptors may not be well developed in 28-day old rats.


In vitro characterization of a gamma-secretase radiotracer in mammalian brain.
Inhibition of gamma-secretase is a potential therapeutic target for Alzheimer's disease (AD). The present studies have characterized the in vitro properties of a radiolabeled small molecule gamma-secretase inhibitor, [3H]compound D (Yan et al., 2004, J. Neurosci.24, 2942-2952) in mammalian brain. [3H]Compound D was shown to bind with nanomolar affinity (Kd = 0.32-1.5 nM) to a single population of saturable sites in rat, rhesus and human brain cortex homogenates, the density of binding sites ranging from 4 to 7 nM across the species. Competition studies with a structurally diverse group of gamma-secretase inhibitors with a wide range of binding affinities showed that the binding affinities of these compounds correlated well with their ability to inhibit gamma-secretase in vitro. Autoradiographic studies showed that the specific binding of [3H]compound D was widely distributed throughout adult rat, rhesus and normal human brain. There did not appear to be any difference in distribution of [3H]compound D specific binding sites in AD cortex compared with control human cortex as measured using tissue section autoradiography, nor any correlation between gamma-secretase binding and plaque burden as measured immunohistochemically. [3H]compound D is a useful tool to probe the expression and pharmacology of gamma-secretase in mammalian brain.


Lysophosphatidic acid stimulates cell growth by different mechanisms in SKOV-3 and Caov-3 ovarian cancer cells: distinct roles for Gi- and Rho-dependent pathways.
Lysophosphatidic acid (LPA) is an autocrine growth signal critical to the initiation and progression of ovarian cancer. In the current study, we investigated the receptors and signaling cascades responsible for mediating LPA-stimulated cell growth in SKOV-3 and Caov-3 ovarian cancer cell lines. Pharmacological inhibitors of distinct LPA and epidermal growth factor receptors, G proteins and kinases were tested for their effect on LPA-stimulated cell growth, MAP kinase activation and Akt activation in SKOV-3 and Caov-3 cells. Distinct agonist pharmacological profiles were observed. Saturated and unsaturated LPA species were equally potent in Caov-3 cells, while saturated LPA was less potent than unsaturated LPA in SKOV-3 cells. Further, the LPA1/LPA3 receptor antagonist Ki16425 was more potent in SKOV-3 cells. The effect of LPA on cell growth in both cell lines was dependent on phosphatidylinositol-3 kinases and MAP kinases. However, LPA-stimulated SKOV-3 cell growth required Gi G proteins, while Caov-3 cell growth was dependent on the Rho effector p160 Rho kinase. Finally, we demonstrated that regulator of G protein signaling proteins significantly regulated Gi-dependent LPA-stimulated cell growth in SKOV-3 cells. LPA-stimulated cell growth is mediated by distinct but overlapping receptors and signaling pathways in these two model ovarian cancer cell lines.


Applications of expert computer systems.
An expert system is a computer program which uses artificial intelligence to make logical decisions on the basis of input data. The rules the system uses to make its decisions are called heuristics which can be provided in the form of IF . . . THEN statements, or they can be learned by the system from examples. The term "expert" is used because the rules or examples come from human experts and the program is considered to have captured their expertise. Currently there are many expert systems in business and medical use but few, if any, are used in optometry. The rule-oriented nature of many ophthalmic procedures suggests a future role for these systems, but their cost-effectiveness may not yet be favorable enough to justify development of expert systems for optometric practice.


Understanding health inequalities for uninsured Americans: a population-wide survey.
Numbers of the uninsured in America have risen in the past few years to more than 40 million people, yet relatively little is known about their health communication behaviors. Data from the 2003 Health Information National Trends Survey (HINTS) were used to analyze the relationship among demographics, health status, health insurance status, online health seeking, and amount of attention paid to various media for health. A random sample of 6,369 Americans indicated several statistically significant differences between the insured and uninsured: the uninsured were more likely younger, less educated, and Hispanic. Findings also indicated that those without health insurance reported being less healthy and more distressed and hold a greater risk perception for cancer, compared with their insured counterparts. Health insurance, when controlling for demographics and health status, explained a statistically significant but small amount of variance in both online health seeking and attention to health messages in various other media.


The Candidate Cancer Gene Database: a database of cancer driver genes from forward genetic screens in mice.
Identification of cancer driver gene mutations is crucial for advancing cancer therapeutics. Due to the overwhelming number of passenger mutations in the human tumor genome, it is difficult to pinpoint causative driver genes. Using transposon mutagenesis in mice many laboratories have conducted forward genetic screens and identified thousands of candidate driver genes that are highly relevant to human cancer. Unfortunately, this information is difficult to access and utilize because it is scattered across multiple publications using different mouse genome builds and strength metrics. To improve access to these findings and facilitate meta-analyses, we developed the Candidate Cancer Gene Database (CCGD, http://ccgd-starrlab.oit.umn.edu/). The CCGD is a manually curated database containing a unified description of all identified candidate driver genes and the genomic location of transposon common insertion sites (CISs) from all currently published transposon-based screens. To demonstrate relevance to human cancer, we performed a modified gene set enrichment analysis using KEGG pathways and show that human cancer pathways are highly enriched in the database. We also used hierarchical clustering to identify pathways enriched in blood cancers compared to solid cancers. The CCGD is a novel resource available to scientists interested in the identification of genetic drivers of cancer.


Baicalein inhibits the migration and invasion of colorectal cancer cells via suppression of the AKT signaling pathway.
The anticancer effect of baicalein has been known for a number of years. However, its anti-metastatic effect and associated mechanisms in colorectal cancer (CRC) remain unclear. The present study investigated the hypothesis that treatment with baicalein may inhibit the proliferation, motility and invasion of human CRC cell lines via regulation of the protein kinase B (AKT) signaling pathway. Baicalein was demonstrated to significantly inhibit the migration and invasion of CRC cells (P=0.01). Additionally, after treatment with baicalein for 24 h, the protein expression levels of matrix metalloproteinase-2 (MMP-2) and MMP-9 in CRC cells were significantly reduced in a dose-dependent manner (P=0.01). Furthermore, treatment with baicalein significantly reduced the expression levels of phosphorylated AKT (P=0.01). In conclusion, baicalein appears to inhibit CRC cell migration and invasion by reducing the expression of MMP-2 and MMP-9 via suppression of the AKT signaling pathway. Thus, baicalein is a potential novel therapeutic agent for patients with CRC.


Intoxication by star fruit (Averrhoa carambola) in six dialysis patients? (Preliminary report)
We observed six cases of patients in a dialysis programme who were apparently intoxicated by ingestion of star fruit. After ingestion of 2-3 fruits or 150-200 ml of the fruit juice, the six patients, who had previously been stable in a regular dialysis programme, developed a variety of symptoms ranging from insomnia and hiccups to agitation, mental confusion and (in one case) death. In preliminary investigations to characterize the hypothetical neurotoxin in the fruit, an extract, when injected intraperitoneally or intracerebroventricularly in rats, provoked persistent convulsions of the tonic-clonic type. It appears that star fruit (Averrhoa carambola) contains an excitatory neurotoxin. Patients with renal failure on conservative or dialysis treatment should be dissuaded from ingestion of the fruit.


[Utility of analytical parameters in the diagnosis of liver disease].
Liver function tests include biochemical parameters (AST, ALT, GGT or Alkaline phosphatase), bilirubin and albumin levels and coagulation tests as prothrombin activity. These tests are commonly used in the routine screening even in symptomatic as in asymptomatic patients, and the right evaluation of the results is of vital importance. Cytolytic elevation in serum aminotransferases: In mild chronic elevation pharmacological toxicity, viral hepatitis, alcoholic and non-alcoholic fatty liver disease and hemochromatosis, should be excluded. Cholestatic elevation os serum enzymes: The first option should be to establish the origin of the alkaline phosphatase elevation, with the evaluation of the GGT levels to confirm the hepatic origin. The next step should be to distinguish the presence of an extrahepatic (biliary obstruction) or intrahepatic (PBC, PSC, drugs, etc) cholestasis, in these cases the most important test should be the abdominal ultrasound, in order to evaluate the biliary system. Hyperbilirubinemia: Non conjugated hyperbilirubinemia (hemolysis, ineffective erythropoiesis, Gilbert or Criggler-Najjar syndromes) and conjugated hyperbilirubinemia, an unusual situation in which Rotor and Dubin-Johnson Syndromes should be considered. The evaluation of albumin and prothrombin levels evaluates the hepatic function per se, allowing to differentiate between acute and chronic diseases. At present, there are not prospective studies to evaluate the efficacy of the liver function tests. To carry out a complete medical history, an appropriate physical examination and the appropriate application of non-invasive diagnostic tests (serology, iron levels, autoimmunity or abdominal ultrasound) allow to perform a right diagnosis in most patients, making more complex tests, including liver biopsy, secondary.


Coupled macroscopic and microscopic scale modeling of fibrillar tissues and tissue equivalents.
Collagen mechanics are crucial to the function and dysfunction of many tissues, including blood vessels and articular cartilage, and bioartificial tissues. Previous attempts to develop computer simulations of collagenous tissue based on macroscopic property descriptions have often been limited in application by the simplicity of the model; simulations based on microscopic descriptions, in contrast, have numerical limitations imposed by the size of the mathematical problem. We present a method that combines the tractability of the macroscopic approach with the flexibility of the microstructural approach. The macroscopic domain is divided into finite elements (as in standard FEM). Each element contains a microscopic scale network. Instead of a stress constitutive equation; the macroscopic problem is distributed over the microscopic scale network and solved in each element to satisfy the weak formulation of Cauchy's stress continuity equation over the macroscopic domain. The combined method scales by order 1.1 as the overall number of degrees of freedom is increased, allowing it to handle larger problems than a direct microstructural approach. Model predictions agree qualitatively with tensile tests on isotropic and aligned reconstituted type I collagen gels.


Feeding the Soul via Creation of a Suborganization to Promote a Sense of Community.
Pharmacy faculty commonly report feeling stressed, overwhelmed, exhausted, and burnt out. Women may be disproportionally impacted by personal and professional demands. The purpose of this commentary is to describe one mechanism for creating a suborganization (Circle) that establishes a supportive community to combat burnout and promote professional fulfillment. This commentary is a description of one American Academy of Colleges of Pharmacy (AACP) Women Faculty Special Interest Group (SIG) Circle. The authors describe how one Circle sought to enhance the well-being of its members through the various domains of the Stanford Model of Professional Fulfillment, including personal resilience, workplace efficiency, and creating a culture of well-being. Circles and similar frameworks may be effective tools for combatting burnout, improving fulfillment, and promoting wellness and well-being among women and other groups of faculty.


Adiposity as a risk factor for inflammatory bowel disease and the mediating effect of metabolic and inflammatory status: A population-based cohort study.
To examine whether general and abdominal adiposity was a risk factor for the new-onset of inflammatory bowel disease (IBD) and the potential mediating effect of metabolic and inflammation status. A total of 492,998 individuals free of IBD recruited from 2006 to 2010 in the UK Biobank were included in our study, with ongoing follow-up linking to the health-related outcome. Multivariable Cox regression models were used to evaluate the associations between general adiposity (body mass index) and abdominal adiposity (waist circumference) and the subsequent risk of IBD and its subtype. We also investigated the potential mediating effects of metabolic and inflammation status by carrying out exploratory mediation analyses. During a median follow-up of 12.5 years, we documented 2954 incident IBD cases (915 Crohn's disease [CD] and 2039 ulcerative colitis). After adjustment for important confounders, body mass index (hazard ratio [HR] highest quintile [Q5] vs. lowest quintile [Q1] = 1.18, 95% confidence interval [CI] 1.04-1.32; P-trend = 0.006) and waist circumference (HR Q5 vs. Q1 = 1.30, 95% CI 1.14-1.49; P-trend <0.001) showed a positive association with the risk of IBD. The associations were partially mediated by metabolic status (24%; 15%), C-reactive protein (36%; 19%) and inflammation score (82%; 46%). Adiposity bore a risk factor for incident IBD, whereas unhealthy metabolism, especially inflammation, seemed to be an important intermediate condition between the association. Our findings provide evidence for possible mechanisms relating adiposity to IBD from an epidemiological perspective, and experimental studies are needed for further demonstration.


A single loss of photosynthesis in the diatom order Bacillariales (Bacillariophyta).
Loss of photosynthesis is a common and often repeated trajectory in nearly all major groups of photosynthetic eukaryotes. One small subset of "apochloritic" diatoms in the genus Nitzschia have lost their ability to photosynthesize and require extracellular carbon for growth. Similar to other secondarily nonphotosynthetic taxa, apochloritic diatoms maintain colorless plastids with highly reduced plastid genomes. Although the narrow taxonomic breadth of apochloritic Nitzschia suggests a single loss of photosynthesis in their common ancestor, previous phylogenetic analyses suggested that photosynthesis was lost multiple times. We analyzed genes from the nuclear, plastid, and mitochondrial genomes for a broad set of taxa to test whether photosynthesis was lost one or multiple times in Bacillariales. We also sequenced and characterized the plastid genome of a nonphotosynthetic Nitzschia species. Phylogenetic analyses showed that genes from all three genetic compartments either supported or failed to reject monophyly of apochloritic Nitzschia species, consistent with a single loss of photosynthesis in this group. The plastid genomes of two apochloritic Nitzschia are highly similar in all respects, indicating streamlining of the plastid genome before the split of these two species. A better understanding of the phylogeny and ecology of apochloritic Nitzschia, together with emerging genomic resources, will help identify the factors that have driven and maintained the loss of photosynthesis in this group of diatoms. Finally, some habitats host diverse communities of co-occurring nonphotosynthetic diatoms, reflecting resource abundance or resource partitioning in ecologically favorable habitats.


Deciphering myeloid-derived suppressor cells: isolation and markers in humans, mice and non-human primates.
In cancer, infection and inflammation, the immune system's function can be dysregulated. Instead of fighting disease, immune cells may increase pathology and suppress host-protective immune responses. Myeloid cells show high plasticity and adapt to changing conditions and pathological challenges. Despite their relevance in disease pathophysiology, the identity, heterogeneity and biology of myeloid cells is still poorly understood. We will focus on phenotypical and functional markers of one of the key myeloid regulatory subtypes, the myeloid derived suppressor cells (MDSC), in humans, mice and non-human primates. Technical issues regarding the isolation of the cells from tissues and blood, timing and sample handling of MDSC will be detailed. Localization of MDSC in a tissue context is of crucial importance and immunohistochemistry approaches for this purpose are discussed. A minimal antibody panel for MDSC research is provided as part of the Mye-EUNITER COST action. Strategies for the identification of additional markers applying state of the art technologies such as mass cytometry will be highlighted. Such marker sets can be used to study MDSC phenotypes across tissues, diseases as well as species and will be crucial to accelerate MDSC research in health and disease.


Transposition of a fungal miniature inverted-repeat transposable element through the action of a Tc1-like transposase.
The mimp1 element previously identified in the ascomycete fungus Fusarium oxysporum has hallmarks of miniature inverted-repeat transposable elements (MITEs): short size, terminal inverted repeats (TIRs), structural homogeneity, and a stable secondary structure. Since mimp1 has no coding capacity, its mobilization requires a transposase-encoding element. On the basis of the similarity of TIRs and target-site preference with the autonomous Tc1-like element impala, together with a correlated distribution of both elements among the Fusarium genus, we investigated the ability of mimp1 to jump upon expression of the impala transposase provided in trans. Under these conditions, we present evidence that mimp1 transposes by a cut-and-paste mechanism into TA dinucleotides, which are duplicated upon insertion. Our results also show that mimp1 reinserts very frequently in genic regions for at least one-third of the cases. We also show that the mimp1/impala double-component system is fully functional in the heterologous species F. graminearum, allowing the development of a highly efficient tool for gene tagging in filamentous fungi.


Molecular evolution: codes, clocks, genes and genomes.
The discoveries, advancements and continuing controversies in the field of molecular evolution are reviewed. Topics summarized are (1) the evolution of the genetic code, (2) gene evolution including the demonstration of homology, estimation of sequence divergence, phylogenetic trees, the molecular clock and the origin of genes and gene families by various genetic mechanisms, and (3) eukaryotic genome evolution, including the highly repeated satellite sequences, the interspersed and potentially mobile repeated sequences and the unique sequence fraction of the genome.


Clinical implications of scapular dyskinesis in shoulder injury: the 2013 consensus statement from the 'Scapular Summit'.
The second international consensus conference on the scapula was held in Lexington Kentucky. The purpose of the conference was to update, present and discuss the accumulated knowledge regarding scapular involvement in various shoulder injuries and highlight the clinical implications for the evaluation and treatment of shoulder injuries. The areas covered included the scapula and shoulder injury, the scapula and sports participation, clinical evaluation and interventions and known outcomes. Major conclusions were (1) scapular dyskinesis is present in a high percentage of most shoulder injuries; (2) the exact role of the dyskinesis in creating or exacerbating shoulder dysfunction is not clearly defined; (3) shoulder impingement symptoms are particularly affected by scapular dyskinesis; (4) scapular dyskinesis is most aptly viewed as a potential impairment to shoulder function; (5) treatment strategies for shoulder injury can be more effectively implemented by evaluation of the dyskinesis; (6) a reliable observational clinical evaluation method for dyskinesis is available and (7) rehabilitation programmes to restore scapular position and motion can be effective within a more comprehensive shoulder rehabilitation programme.


Renal clearance of glutathione measured in rats pretreated with inhibitors of glutathione metabolism.
Renal clearance measurements were performed in rats pretreated with various inhibitors of glutathione metabolism. Approximately 97% of the renal gamma-glutamyltranspeptidase was inactivated by infusion of AT-125. Within 4.5 h arterial plasma concentration and urinary excretion of glutathione increased from 4 microM and 1.2 nmol/h to 27 microM and 3,900 nmol/h, respectively. The ratio of excreted to filtered glutathione increased from less than 0.01 to 1. When renal glutathione was decreased to 35% of normal by pretreating rats with D,L-buthionine-S,R-sulfoximine, the subsequent inactivation of the transpeptidase caused only a two-fold increase in arterial plasma glutathione, and urinary excretion increased to only 70% of the filtered load. The filtered load of glutathione was reduced by intravenous infusion of purified gamma-glutamyltranspeptidase. At 1 to 3 h after infusion, arterial glutathione decreased to 0.3 microM. Under these conditions, the amount of glutathione excreted by an AT-125-treated rat was 40-fold greater than the filtered load. When similar experiments were carried out in rats that were pretreated with D,L-buthionine-S,R-sulfoximine and AT-125, urinary excretion of glutathione was decreased 10-fold, but it still exceeded the amount filtered. These results firmly establish that apical secretion contributes to the glutathione catabolized in the tubular lumen.


Medication-Wide Study: Exploring Medication Use 10 Years Before a Diagnosis of Inflammatory Bowel Disease.
There is growing interest in the prediagnostic phase of inflammatory bowel disease (IBD) and in the overlap of IBD with other diseases. We described and compared use of any prescription medication between individuals with and without IBD in a 10-year period preceding diagnosis. Based on cross-linked nationwide registers, we identified 29,219 individuals diagnosed with IBD in Denmark between 2005 and 2018 and matched to 292,190 IBD-free individuals. The primary outcome was use of any prescription medication in years 1-10 before IBD diagnosis/matching date. Participants were considered as medication users if they redeemed ≥1 prescription for any medication in the World Health Organization Anatomical Therapeutic Chemical (ATC) main groups or subgroups before diagnosis/matching. The IBD population had a universally increased use of medications compared with the matched population before IBD diagnosis. At 10 years before diagnosis, the proportion of users was 1.1-fold to 1.8-fold higher in the IBD population in 12 of 14 ATC main groups of medication ( P -value < 0.0001). This applied across age, sex, and IBD subtypes, although it was the most pronounced for Crohn's disease (CD). Two years before diagnosis, the IBD population had a steep increase in medication use for several organ systems. When analyzing therapeutic subgroups of medication, the CD population exhibited 2.7, 2.3, 1.9, and 1.9 times more users of immunosuppressants, antianemic preparations, analgesics, and psycholeptics, respectively, than the matched population 10 years before diagnosis ( P -value < 0.0001). Our findings demonstrate universally increased medication use years before IBD, especially CD, diagnosis and indicates multiorgan involvement in IBD.


Adenosine and opioid receptor-mediated cardioprotection in the rat: evidence for cross-talk between receptors.
The relative roles of free-radical production, mitochondrial ATP-sensitive K+ (mitoKATP) channels and possible receptor cross-talk in both opioid and adenosine A1 receptor (A1AR) mediated protection were assessed in a rat model of myocardial infarction. Sprague-Dawley rats were subjected to 30 min of occlusion and 90 min of reperfusion. The untreated rats exhibited an infarct of 58.8 +/- 2.9% [infarct size (IS)/area at risk (AAR), %] at the end of reperfusion. Pretreatment with either the nonselective opioid receptor agonist morphine or the selective A1AR agonist 2-chloro-cyclopentyladenosine (CCPA) dramatically reduced IS/AAR to 41.1 +/- 2.2% and 37.9 +/- 5.5%, respectively (P < 0.05). Protection afforded by either morphine or CCPA was abolished by the reactive oxygen species scavenger N-(2-mercaptopropionyl)glycine or the mitoKATP channel blocker 5-hydroxydecanoate. Both morphine- and CCPA-mediated protection were attenuated by the selective A1AR antagonist 1,3-dipropyl-8-cyclopentylxanthine and the selective delta1-opioid receptor (DOR) antagonist 7-benzylidenealtrexone. Simultaneous administration of morphine and CCPA failed to enhance the infarct-sparing effect of either agonist alone. These data suggest that both DOR and A1AR-mediated cardioprotection are mitoKATP and reactive oxygen species dependent. Furthermore, these data suggest that there are converging pathways and/or receptor cross-talk between A1AR- and DOR-mediated cardioprotection.


Comparison of deposited surface area of airborne ultrafine particles generated from two welding processes.
This article describes work performed on the assessment of the levels of airborne ultrafine particles emitted in two welding processes metal-active gas (MAG) of carbon steel and friction-stir welding (FSW) of aluminium in terms of deposited area in alveolar tract of the lung using a nanoparticle surface area monitor analyser. The obtained results showed the dependence from process parameters on emitted ultrafine particles and clearly demonstrated the presence of ultrafine particles, when compared with background levels. The obtained results showed that the process that results on the lower levels of alveolar-deposited surface area is FSW, unlike MAG. Nevertheless, all the tested processes resulted in important doses of ultrafine particles that are to be deposited in the human lung of exposed workers.


PlexinD1 Is a Novel Transcriptional Target and Effector of Notch Signaling in Cancer Cells.
The secreted semaphorin Sema3E controls cell migration and invasiveness in cancer cells. Sema3E-receptor, PlexinD1, is frequently upregulated in melanoma, breast, colon, ovarian and prostate cancers; however, the mechanisms underlying PlexinD1 upregulation and the downstream events elicited in tumor cells are still unclear. Here we show that the canonical RBPjk-dependent Notch signaling cascade controls PlexinD1 expression in primary endothelial and cancer cells. Transcriptional activation was studied by quantitative PCR and promoter activity reporter assays. We found that Notch ligands and constitutively activated intracellular forms of Notch receptors upregulated PlexinD1 expression; conversely RNAi-based knock-down, or pharmacological inhibition of Notch signaling by gamma-secretase inhibitors, downregulated PlexinD1 levels. Notably, both Notch1 and Notch3 expression positively correlates with PlexinD1 levels in prostate cancer, as well as in other tumor types. In prostate cancer cells, Sema3E-PlexinD1 axis was previously reported to regulate migration; however, implicated mechanisms were not elucidated. Here we show that in these cells PlexinD1 activity induces the expression of the transcription factor Slug, downregulates E-cadherin levels and enhances cell migration. Moreover, our mechanistic data identify PlexinD1 as a pivotal mediator of this signaling axis downstream of Notch in prostate cancer cells. In fact, on one hand, PlexinD1 is required to mediate cell migration and E-cadherin regulation elicited by Notch. On the other hand, PlexinD1 upregulation is sufficient to induce prostate cancer cell migration and metastatic potential in mice, leading to functional rescue in the absence of Notch. In sum, our work identifies PlexinD1 as a novel transcriptional target induced by Notch signaling, and reveals its role promoting prostate cancer cell migration and downregulating E-cadherin levels in Slug-dependent manner. Collectively, these findings suggest that Notch-PlexinD1 signaling axis may be targeted to impair prostate cancer cell invasiveness and metastasis.


First Report of Anthracnose Stalk Rot of Maize Caused by Colletotrichum graminicola in Switzerland.
Maize stem samples exhibiting symptoms of anthracnose were collected from a field near Zurich, Switzerland, in September of 2012 and were sent to the fungal genetics laboratory, Centro Hispano-Luso de Investigaciones Agrarias (CIALE) at the University of Salamanca, Spain, for further analysis. The stem samples exhibited glossy, black, and irregularly shaped lesions. Tissue samples, approximately 5 mm2, were dissected from below the epidermis. The tissue samples were surface disinfested for 1 min in 20% sodium hypochlorite and cultured on one half strength acidified PDA supplemented with ampicillin (2). Monoconidial isolates from three different stems were grown on potato dextrose agar (PDA) and had dark gray aerial mycelium with orange spore masses. Conidia were falcate, slightly curved, tapered toward the tips with an average length of 31.77 μm and an average width of 4.76 μm and produced in acervuli with setae, consistent with descriptions of C. graminicola Ces. Wils. Conidial suspensions were prepared for each isolate, and were inoculated onto the leaves of 2-week-old maize plants by laying the plants horizontally in a tray (in pots with their root systems intact) and placing 7.5-μl droplets of a 106 conidial suspension on the leaf surface. The trays were covered and plants were incubated overnight at 23°C. The plants were then returned to their upright position and grown in a growth chamber at 25°C with a 12-h light cycle (3). After 6 days, the inoculated plant leaves exhibited lesions that were elongated and irregularly shaped with necrotic centers and chlorotic margins. The water-inoculated controls did not show symptoms. Microscopic examination revealed the production of conidia on the surface of the leaves, identical to the original isolates. Genomic DNA was extracted using the protocol of Baek and Kenerley (1). A region of the ribosomal DNA repeat was amplified and sequenced using the universal primers ITS4 and ITS5. The resulting sequences were 100% identical to each other and 100% identical to C. graminicola sequences in GenBank. One representative sequence was deposited in GenBank under accession no. KF597538. The 100 most similar sequences in GenBank were used to construct a phylogenetic tree using the neighbor-joining method. The phylogenetic analysis revealed that the isolates clustered within the C. graminicola clade, consistent with their identification as C. graminicola. To our knowledge, this is the first report of anthracnose on maize caused by C. graminicola in Switzerland. Previous reports have demonstrated that the pathogen exists in neighboring countries Germany and France. References: (1) J.-M. Baek and C. M. Kenerley. Fungal Genet. Biol. 23:34, 1998. (2) S. A. Sukno et al. Appl. Environ. Microbiol. 74:823, 2008. (3) W. A. Vargas et al. Plant Physiol. 158:1342, 2012.


Some guiding principles in emerging models of voice therapy for children.
Voice disorders are the most common communication disorder across the life span, affecting more than five million school-aged children annually in the United States alone. Although such disorders are commonly associated with benign pathology, medically malicious conditions cannot be ruled out without proper medical examination by an otolaryngologist. Many children with voice disorders are otherwise healthy. However, some are not. Moreover, children with voice problems of any origin often experience distinctly negative consequences in quality of life and academic participation as a result of them. Regrettably, this population is badly underserved by speech-language pathology. In this article, possible reasons are suggested, among them the relative unattractiveness of traditional therapy that emphasizes voice conservation using a Skinnerian paradigm. Alternatives are presented in terms of foundational principles of biomechanics, biology, and learning and factors affecting children's compliance with health care directives, which may help to shape the next generation of pediatric voice therapies. The article concludes with discussion of challenges surrounding voice therapy in the school setting and possible solutions.


Patient Activation of Persons With Opioid Use Disorder in Intensive Outpatient Treatment.
High patient activation is associated with a variety of positive health outcomes. Although increasing patient activation in persons with opioid use disorder (OUD) in intensive outpatient treatment (IOT) programs may increase engagement and improve outcomes, little is known about how patient activation is manifested in these programs. To describe types of instances in which persons play an active role in their IOT or show self-determination in their recovery generally (patient activation) and types of instances in which they play a passive role in their IOT or have recovery directed by others (patient nonactivation). A qualitative descriptive study using data from a larger grounded theory study was conducted. Interviews were completed with 14 persons with OUD who attended an IOT program within a large health care system in the Midwest. Content analysis was used to create a typology of instances of patient activation or nonactivation in participants' IOT experiences. Six types of instances were identified: (1) making and enacting one's own treatment decisions, (2) actively collaborating with staff, (3) self-determining one's disclosure in groups, (4) making a commitment to treatment, (5) taking responsibility for one's recovery, and (6) taking actions to avoid return to use. Patient activation is multidimensional and plays a salient role in IOT experiences. IOT staff should engage patients as active participants in their treatment and encourage mutual goal-setting and shared-decision but should be aware that some approaches used too early in treatment may impede recovery.


Repression and catabolite repression of the lactose operon of Staphylococcus aureus.
The lacR gene encodes the repressor of the lactose operon of S. aureus. The nucleotide sequence of this gene and the promoter-operator region of the operon are reported. The lacR gene encodes a protein with a molecular weight of 28,534. This protein was found to share sequence homology with the DeoR protein, the repressor of the E. coli deoxyribonucleotide operon. Directly and invertedly repeated sequences were found associated with the promoter for the structural genes of the operon. These sequences were examined by site-directed mutagenesis and found to be important in repressor binding and in the binding of a catabolite repressor. Evidence is presented in support of a model for catabolite repression of the operon which involves a negative-acting transcriptional regulator which binds to the promoter region of the operon and prevents transcription.


Depression and anxiety symptoms moderate the relation between negative reinforcement smoking outcome expectancies and nicotine dependence.
Smoking reinforcement expectancies-expectations that smoking modulates mood-can be powerful motivators to smoke, resulting in increased nicotine dependence. The impact of smoking reinforcement expectancies on nicotine dependence may be particularly strong in individuals with increased mood or anxiety symptoms because they may be more likely to act on expectancies with smoking behavior in order to offset their affective symptoms. This study examined levels of emotional symptom dimensions as moderators of the relation between positive and negative smoking reinforcement expectancies and nicotine dependence severity in a community sample. In a cross-sectional design, 317 daily cigarette smokers (215 men) completed self-report measures of smoking reinforcement expectancies, mood and anxiety symptoms, and nicotine dependence. Increasing levels of negative affect and anxiety symptoms strengthened associations between negative reinforcement smoking expectancies and nicotine dependence severity (moderation effects; (βs > .13; ps < .03) but did not moderate relations between positive reinforcement expectancies and dependence. Anhedonia did not moderate relations involving either positive or negative reinforcement smoking expectancies. Distinct components of anxiety and depressive symptoms interact differently with smoking reinforcement expectancies. Emotional symptoms characterized by excesses in aversive (but not deficits in appetitive) functioning may amplify tendencies to compulsively act on negative reinforcement expectancies by smoking. Cessation treatments that target negative reinforcement expectancies may be particularly salient for emotionally distressed smokers.


Effect of Intraoperative Low Tidal Volume vs Conventional Tidal Volume on Postoperative Pulmonary Complications in Patients Undergoing Major Surgery: A Randomized Clinical Trial.
In patients who undergo mechanical ventilation during surgery, the ideal tidal volume is unclear. To determine whether low-tidal-volume ventilation compared with conventional ventilation during major surgery decreases postoperative pulmonary complications. Single-center, assessor-blinded, randomized clinical trial of 1236 patients older than 40 years undergoing major noncardiothoracic, nonintracranial surgery under general anesthesia lasting more than 2 hours in a tertiary hospital in Melbourne, Australia, from February 2015 to February 2019. The last date of follow-up was February 17, 2019. Patients were randomized to receive a tidal volume of 6 mL/kg predicted body weight (n = 614; low tidal volume group) or a tidal volume of 10 mL/kg predicted body weight (n = 592; conventional tidal volume group). All patients received positive end-expiratory pressure (PEEP) at 5 cm H2O. The primary outcome was a composite of postoperative pulmonary complications within the first 7 postoperative days, including pneumonia, bronchospasm, atelectasis, pulmonary congestion, respiratory failure, pleural effusion, pneumothorax, or unplanned requirement for postoperative invasive or noninvasive ventilation. Secondary outcomes were postoperative pulmonary complications including development of pulmonary embolism, acute respiratory distress syndrome, systemic inflammatory response syndrome, sepsis, acute kidney injury, wound infection (superficial and deep), rate of intraoperative need for vasopressor, incidence of unplanned intensive care unit admission, rate of need for rapid response team call, intensive care unit length of stay, hospital length of stay, and in-hospital mortality. Among 1236 patients who were randomized, 1206 (98.9%) completed the trial (mean age, 63.5 years; 494 [40.9%] women; 681 [56.4%] undergoing abdominal surgery). The primary outcome occurred in 231 of 608 patients (38%) in the low tidal volume group compared with 232 of 590 patients (39%) in the conventional tidal volume group (difference, -1.3% [95% CI, -6.8% to 4.2%]; risk ratio, 0.97 [95% CI, 0.84-1.11]; P = .64). There were no significant differences in any of the secondary outcomes. Among adult patients undergoing major surgery, intraoperative ventilation with low tidal volume compared with conventional tidal volume, with PEEP applied equally between groups, did not significantly reduce pulmonary complications within the first 7 postoperative days. ANZCTR Identifier: ACTRN12614000790640.