Further characterization of porcine Brachyspira hyodysenteriae isolates with decreased susceptibility to tiamulin.
Brachyspira hyodysenteriae is the causative agent of swine dysentery, a severe diarrhoeal disease in pigs. Few drugs are available to treat the disease, owing to both antimicrobial resistance and withdrawal of drugs authorized for use in pigs. Tiamulin is the drug of choice in many countries, but isolates with decreased susceptibility have recently been reported. The mechanism of tiamulin resistance in B. hyodysenteriae is not known and this facet is essential to understand the dissemination of the trait. To study the resistance epidemiology of B. hyodysenteriae, further characterization of a set of isolates from Germany (n = 16) and the UK (n = 6) with decreased susceptibility to tiamulin was performed. The relatedness between the isolates was studied by comparing PFGE patterns, and the in vitro susceptibility to five other antimicrobials (aivlosin, doxycycline, salinomycin, chloramphenicol and avilamycin) was also determined. For comparison of the antimicrobial-susceptibility pattern, Swedish (n = 20) and British (n = 4) tiamulin-susceptible isolates were tested. The German isolates represented several different PFGE patterns, indicating that tiamulin usage has been sufficient to select clones with decreased tiamulin susceptibility at different farms in Germany. The PFGE pattern for the six British isolates with decreased tiamulin susceptibility was identical to that of the German isolates, and they had a similar antimicrobial-susceptibility pattern, except for resistance to aivlosin, which was only found in a few German isolates. No other co-resistance with tiamulin was found.


Environmental policy. Counting the cost of deforestation.
To halt the dramatic alteration in our climate, there must be a reduction in the emission of greenhouse gases. As Bonnie and colleagues discuss in their Perspective, conservation of forests will increase carbon sequestration and decrease greenhouse gas emissions. In this issue, a cost-benefit analysis by Kremen et al. demonstrates the benefits of forest conservation on a local and global scale.


Analysis by high throughput sequencing of Specific Pathogen Free eggs.
Specific Pathogen Free (SPF) embryonated eggs are used for the production of many veterinary and human vaccines. We have used High Throughput Sequencing to screen allantoic fluids and embryos for the presence of encapsidated viral genomes and viral transcripts, respectively. SPF eggs from two different producers were tested. We evidenced sequences corresponding to known endogenous retroviruses and sequences of Avian Leukosis Virus, but no sequence that might suggest a productive infection of eggs with a virus even distant from known viruses. Our results strongly suggest that SPF eggs such as those used for this study represent a safe substrate for the production of vaccines.


Microfluidics for in vitro biomimetic shear stress-dependent leukocyte adhesion assays.
Recruitment of leukocytes from blood to tissues is a multi-step process playing a major role in the activation of inflammatory responses. Tethering and rolling of leukocytes along the vessel wall, followed by arrest and transmigration through the endothelium result from chemoattractant-dependent signals, inducing adhesive and migratory events. Shear forces exerted by the blood flow on leukocytes induce rolling via selectin-mediated interactions with endothelial cells and increase the probability of leukocytes to engage their chemokine receptors, facilitating integrin activation and consequent arrest. Flow-derived shear forces generate mechanical stimuli concurring with biochemical signals in the modulation of leukocyte-endothelial cell interactions. In the last few years, a host of in vitro studies have clarified the biochemical adhesion cascade and the role of shear stress in leukocyte extravasation. The limitation of the static environment in Boyden devices has been overcome both by the use of parallel-plate flow chambers and by custom models mimicking the in vivo conditions, along with widespread microfluidic approaches to in vitro modeling. These devices create an in vitro biomimetic environment where the multi-step transmigration process can be imaged and quantified under mechanical and biochemical controlled conditions, including fluid dynamic settings, channel design, materials and surface coatings. This paper reviews the technological solutions recently proposed to model, observe and quantify leukocyte adhesion behavior under shear flow, with a final survey of high-throughput solutions featuring multiple parallel assays as well as thorough and time-saving statistical interpretation of the experimental results.


Structural basis for the binding of SNAREs to the multisubunit tethering complex Dsl1.
Multisubunit-tethering complexes (MTCs) are large (250 to >750 kDa), conserved macromolecular machines that are essential for soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-mediated membrane fusion in all eukaryotes. MTCs are thought to organize membrane trafficking by mediating the initial long-range interaction between a vesicle and its target membrane and promoting the formation of membrane-bridging SNARE complexes. Previously, we reported the structure of the yeast Dsl1 complex, the simplest known MTC, which is essential for coat protein I (COPI) mediated transport from the Golgi to the endoplasmic reticulum (ER). This structure suggests how the Dsl1 complex might tether a vesicle to its target membrane by binding at one end to the COPI coat and at the other to ER-associated SNAREs. Here, we used X-ray crystallography to investigate these Dsl1-SNARE interactions in greater detail. The Dsl1 complex comprises three subunits that together form a two-legged structure with a central hinge. We found that distal regions of each leg bind N-terminal Habc domains of the ER SNAREs Sec20 (a Qb-SNARE) and Use1 (a Qc-SNARE). The observed binding modes appear to anchor the Dsl1 complex to the ER target membrane while simultaneously ensuring that both SNAREs are in open conformations, with their SNARE motifs available for assembly. The proximity of the two SNARE motifs, and therefore their ability to enter the same SNARE complex, will depend on the relative orientation of the two Dsl1 legs. These results underscore the critical roles of SNARE N-terminal domains in mediating interactions with other elements of the vesicle docking and fusion machinery.


Accentuated Eccentric Loading and Cluster Set Configurations in the Back Squat: A Kinetic and Kinematic Analysis.
Wagle, JP, Cunanan, AJ, Carroll, KM, Sams, ML, Wetmore, A, Bingham, GE, Taber, CB, DeWeese, BH, Sato, K, Stuart, CA, and Stone, MH. Accentuated eccentric loading and cluster set configurations in the back squat: a kinetic and kinematic analysis. J Strength Cond Res 35(2): 420-427, 2021-This study examined the kinetic and kinematic differences between accentuated eccentric loading (AEL) and cluster sets in trained male subjects (age = 26.1 ± 4.1 years, height = 183.5 ± 4.3 cm, body mass = 92.5 ± 10.5 kg, and back squat to body mass ratio = 1.8 ± 0.3). Four load condition sessions consisted of traditionally loaded (TL) "straight sets," TL cluster (TLC) sets, AEL cluster (AEC) sets, and AEL "straight sets" where only the first repetition had eccentric overload (AEL1). An interrepetition rest interval of 30 seconds was prescribed for both TLC and AEC. Concentric intensity for all load conditions was 80% 1 repetition maximum (1RM). Accentuated eccentric loading was applied to repetitions using weight releasers with total eccentric load equivalent to 105% of concentric 1RM. Traditionally loaded cluster had statistically greater concentric outputs than TL. Furthermore, statistically greater eccentric and concentric outputs were observed during AEC compared with TL with the exception of peak power. Statistically greater concentric characteristics were observed in TLC compared with AEL1, but statistically greater eccentric outputs were observed in AEL1. In the 2 cluster set conditions, statistically greater concentric rate of force development (RFDCON) (d = 0.470, p < 0.001) and average velocity (vavg) (d = 0.560, p < 0.001) in TLC compared with AEC were observed. However, statistically greater eccentric work (WECC) (d = 2.096, p < 0.001) and eccentric RFD (RFDECC) (d = 0.424, p < 0.001) were observed in AEC compared with TLC. Overall, eccentric overload demonstrated efficacy as a means of increasing eccentric work and RFD, but not as a means of potentiating concentric output. Finally, interrepetition rest seems to have the largest influence on concentric power output and RFD.


Clinical and molecular characterization of 112 single-center patients with Neurofibromatosis type 1.
The aim of this retrospective study was to define clinical and molecular characteristics of a large sample of neurofibromatosis type 1 (NF1) patients, as well as to evaluate mutational spectrum and genotype-phenotype correlation. NF1 is a relatively common neurogenetic disorder (1:2500-1:3000 individuals). It is caused by mutations of the NF1 gene on chromosome 17ql1.2, with autosomal dominant pattern of inheritance and wide phenotypical variability. Café-au-lait spots (CALs), cutaneous and/or subcutaneous neurofibromas (CNFs/SCNFs), skinfold freckling, skeletal abnormalities, Lisch nodules of the iris and increased risk of learning and intellectual disabilities, as well as tumors of the nervous system and other organs are its main clinical features. The preliminary group collected 168 subjects with clinical suspicion of NF1. They were evaluated following the National Institutes of Health (NIH) criteria for NF1, revised by Gutmann et al. 1997, integrated for 67 of them by molecular testing. According to these references, 112 of 168 patients were diagnosed as NF1. The sample was characterized by an equal sex ratio (57 males, 55 females) and age distribution ranging from 10 days to 60 years of age (mean age, 13 years). A wide spectrum of clinical features has been observed in our patients. Mutational analysis resulted positive in 51 cases (76%). Twenty-four mutations detected in our cohort have not been reported to date. This study may contribute to a better definition of genotypic and phenotypic features of NF1 patients, with respect to further insights into the clinical characterization of the disease. In addition, an amplification of the spectrum of mutations in the NF1 gene has been documented.


The relationship of living arrangements and depressive symptoms among older adults in sub-Saharan Africa.
Older adults in sub-Saharan Africa are increasingly facing the twin challenges of reduced support from their adult children and taking on new roles caring for orphans and vulnerable children. How these changes affect the mental health of older adults is largely unknown. We use data from the 2002-2003 World Health Surveys for 15 countries in sub-Saharan Africa to examine whether older adults who may be lacking adequate support through living alone or in skipped-generation households are at an increased risk of depressive symptoms compared to those living with at least one working-age adult. Using meta-regression, we also examine whether heterogeneity across countries in the prevalence of depressive symptoms or in the association between living arrangements and depressive symptoms is associated with HIV/AIDS prevalence and national economic status. The pooled prevalence of depressive symptoms among older adults was 9.2%. Older adults living alone had a 2.3% point higher predicted prevalence of depressive symptoms compared to individuals living with at least one working-age adult (95% confidence interval: 0.2%, 4.4%). None of the country characteristics examined explained heterogeneity across countries in the relationship between living arrangements and depressive symptoms. However, there was some evidence suggesting a positive association between depressive symptom prevalence and the severity of a country's HIV/AIDS epidemic. As depressive symptoms are known to be predictive of poor quality of life and increased mortality, it is important to address how health and social policies can be put in place to mitigate the potentially detrimental effects of solitary living on the mental health of older persons in sub-Saharan Africa.


Visit-to-visit variability of blood pressure and cardiovascular disease and all-cause mortality: a systematic review and meta-analysis.
Visit-to-visit variability of blood pressure (BP) has been associated with cardiovascular disease (CVD) and mortality in some but not all studies. We conducted a systematic review and meta-analysis to examine the association between visit-to-visit variability of BP and CVD and all-cause mortality. Medical databases were searched through June 4, 2014, for studies meeting the following eligibility criteria: adult participants; BP measurements at ≥3 visits; follow-up for CVD, coronary heart disease, stroke, or mortality outcomes; events confirmed via database, death certificate, or event ascertainment committee; and adjustment for confounders. Data were extracted by 2 reviewers and pooled using a random-effects model. Overall, 8870 abstracts were identified of which 37 studies, representing 41 separate cohorts, met inclusion criteria. Across studies, visit-to-visit variability of systolic BP and diastolic BP showed significant associations with outcomes in 181 of 312 (58.0%) and 61 of 188 (32.4%) analyses, respectively. Few studies provided sufficient data for pooling risk estimates. For each 5 mm Hg higher SD of systolic BP, the pooled hazard ratio for stroke across 7 cohorts was 1.17 (95% confidence interval [CI], 1.07-1.28), for coronary heart disease across 4 cohorts was 1.27 (95% CI, 1.07-1.51), for CVD across 5 cohorts was 1.12 (95% CI, 0.98-1.28), for CVD mortality across 5 cohorts was 1.22 (95% CI, 1.09-1.35), and for all-cause mortality across 4 cohorts was 1.20 (95% CI, 1.05-1.36). In summary, modest associations between visit-to-visit variability of BP and CVD and all-cause mortality are present in published studies. However, these findings are limited by the small amount of data available for meta-analysis.


The development of expertise in dermatology.
To examine the development of expertise in dermatology, accuracy of diagnosis and response times of subjects at five levels of expertise were assessed. A total of 100 slides, 2 typical and 3 atypical slides from each of 20 common skin disorders, were presented to six subjects at each of the following levels: second-year preclinical medical students, final year medical students, residents in family medicine, general practitioners, and dermatologists. Accuracy of diagnosis rose from 21% for medical students to 87% for dermatologists. Correct diagnosis was associated with a decrease in response time with expertise, whereas errors were associated with a dramatic increase in response time, and was slower than correct response times at all levels, suggesting that errors do not result predominantly from carelessness or speed. Typical slides accounted for a constant proportion of diagnostic errors at all higher levels of expertise, and experts continued to make a significant proportion of errors on slides shown to be relatively easy for residents. The results are shown to be at variance with any model that equates expertise with the mastery of complex rules, but they are consistent with models of expertise that propose that expertise is equated with a rapid "pattern-recognition" process, and errors result from unintended confusion with previous similar examples.


Survey of Immunization Practices in Patients With Inflammatory Bowel Disease Among Pediatric Gastroenterologists.
We aimed to determine vaccination practices of pediatric gastroenterologists, as well as barriers to following immunization guidelines in patients with inflammatory bowel disease. Institutions listed in the North American Society for Pediatric Gastroenterology, Hepatology, and Nutrition clinical research registry and/or ImproveCareNow were contacted. A total of 657 physicians from 129 institutions were asked to complete a 16-question electronic survey. A total of 178 physicians (27.1%) responded, of whom 55% were male and 83.1% practiced in an academic setting. A total of 11 physicians (6.2%) do not routinely assess vaccination status, whereas 63.5% assess at the time of diagnosis, 29.8% at "well" visits, and 44.4% before initiating immunosuppression. At diagnosis, 51.1% verbally inquire about immunization status, 30.9% obtain records, and 9.0% obtain serology.The influenza (78%), hepatitis B (84%), and varicella (82%) vaccines were most frequently assessed. Fewer than 55.5% of physicians reviewed other vaccines. Physicians using a reminder mechanism were more likely to review immunizations at established visits (41.1% vs 20.8%), and before transfer to an adult gastroenterologist (14.4% vs 2.6%). Lack of coordination of care with primary care practitioners (41%), poor access to immunization records (36%), and inability to offer vaccinations in their immediate area (55%) are barriers to vaccination. Only 28% believed that primary care practitioners were solely responsible for immunizations. There is practice variation among pediatric gastroenterologists in assessment of immunizations in patients with inflammatory bowel disease, including the specific vaccines assessed, and timing and method of assessment. Inability to coordinate care, access immunization records, and offer vaccines through their medical practice are barriers to adhering to immunization guidelines.


H5N1 influenza questionnaire: are Scottish emergency departments ready to isolate and treat a possible index case?
H5N1 has emerged as a highly pathogenic strain of avian flu. Cases of bird-to-human transmission have occurred and raised concerns that human-to-human transmission may be possible. Therefore, it is vitally important to isolate any index case arising from bird-to-human transmission. Questionnaires were sent out to emergency departments throughout Scotland seeking information on the availability of facilities and supplies necessary to treat and prevent onward infection from a suspected index case of human H5N1 infection. They were specifically asked about side rooms for isolation and supplies of FFP3 masks, lysis buffer and oseltamivir (Tamiflu).


A two-year review of suicide ideation assessments among medical, nursing, and pharmacy students.
Suicide is the 10th leading cause of death in the U.S. and has increased in prevalence during the past 15 years. Patients who attempt suicide are more likely to have contact with their primary care provider than a mental health provider in the month before attempting suicide, highlighting the need for competency in suicide ideation (SI) assessment. The Communications Skills for Health Professionals is an interprofessional course involving first-year medical, nursing, and pharmacy students. Specific instruction regarding assessment of SI was delivered through an online module and later practiced by students with standardized patients (SP). A final Objective Structured Clinical Examination featured an SP with depression, but without SI, though an assessment of SI was indicated. Three hundred fifty six interviews were reviewed and 55.1% (196/356) of students assessed for SI. Across professions, 65.5% (93/142) of medical students, 52.5% (32/61) of nursing students, and 46.4% (71/153) of pharmacy students performed an assessment. Medical students' SI assessment was highest across the groups (p = 0.001), while pharmacy students' SI assessment was lowest (p = 0.004). Results suggest that additional educational strategies should be developed and implemented to increase SI assessment performance in all professions, but especially in pharmacy students.


Comorbidity between post-traumatic stress disorder and borderline personality disorder: a review.
Traditionally, the presence of post-traumatic stress disorder (PTSD) in subjects diagnosed with borderline personality disorder (BPD) has been the object of scant empirical research. The clarification of issues related to the different areas of study for this comorbidity is not only significant from a theoretical point of view but also relevant for clinical practice. The aim of this review is to describe the main theoretical findings and research conclusions about the comorbidity between PTSD and BPD. A literature review was carried out via PubMed and PsycINFO for the period between 1990 and September 2013. The descriptors used were 'post-traumatic stress disorder', 'borderline personality disorder', 'PTSD', 'complex PTSD' and 'BPD'. Epidemiological studies show that the risk of PTSD among BPD subjects is not regularly higher than in subjects with other personality disorders. Furthermore, there is no conclusive evidence about the main aetiopathogenic mechanism of this comorbidity, either of one disorder being a risk factor for the other one or of common underlying variables. Concerning comparative studies, several studies with PTSD-BPD subjects have found a higher severity of psychopathology and psychosocial impairment than in BPD subjects. With regard to nosological status, the main focus of controversy is the validation of 'complex PTSD', a clinical entity which may comprise a subgroup of PTSD-BPD subjects. With regard to treatment, there are preliminary evidences for the efficient treatment of psychopathology in both PTSD and BPD. These findings are remarkable for furthering the understanding of the link between PTSD and BPD and their implications for treatment. The results of this review are discussed, including methodological constraints that hinder external validity and consistency of referred findings.


Contrasting population and life history responses of a young morph-pair of European whitefish to the invasion of a specialised coregonid competitor, vendace.
Invasions of non-native species represent a global problem of great scientific interest. Here we study in detail the response in population and life history characteristics of closely related native species, with divergent habitat preferences, that are impacted by an invading species over a sufficient time period to allow a new stable state to become established. A time series of 20 years starting at the first occurrence of the invader (vendace Coregonus albula (L.)) allows exploration of the long term population and life history response of two ecologically, morphologically, and genetically different native sympatric morphs (DR- and SR-) of congeneric whitefish C. lavaretus (L.). The whitefish morphs are taxonomically equally related to the invading vendace, but only the planktivorous DR-whitefish share its pelagic niche. We would expect that the ecological differences between the whitefish morphs may be used as a predictor of competitive effects. Vendace exhibited an initial boom-and-bust development, and has continued to fluctuate in density. The responses of the pelagic DR-whitefish were: i) an immediate habitat shift, ii) a subsequent population decline caused by increased annual mortality, and iii) a new stable state at a lower density and apparently relaxed competition. The ecologically more distant benthivorous SR-whitefish also showed significant, but a much more limited response during this process, indicating damped indirect interactions through the food-web. This long-term case-study found that in two native eco-species equally related to the invader, only one of the eco-species was highly affected. Direct competition for resources is obviously important for species interactions, whereas the taxonomic relatedness per se seems to offer little predictive power for invasion effects.


Presence of nonhemolytic pneumolysin in serotypes of Streptococcus pneumoniae associated with disease outbreaks.
Pneumolysin is an important virulence factor of the human pathogen Streptococcus pneumoniae. Sequence analysis of the ply gene from 121 clinical isolates of S. pneumoniae uncovered a number of alleles. Twenty-two strains were chosen for further analysis, and 14 protein alleles were discovered. Five of these had been reported previously, and the remaining 9 were novel. Cell lysates were used to determine the specific hemolytic activities of the pneumolysin proteins. Six strains showed no hemolytic activity, and the remaining 16 were hemolytic, to varying degrees. We report that the nonhemolytic allele reported previously in serotype 1, sequence type (ST) 306 isolates is also present in a number of pneumococcal isolates of serotype 8 that belong to the ST53 lineage. Serotype 1 and 8 pneumococci are known to be associated with outbreaks of invasive disease. The nonhemolytic pneumolysin allele is therefore associated with the dominant clones of outbreak-associated serotypes of S. pneumoniae.


Co-assembly of Kv4 {alpha} subunits with K+ channel-interacting protein 2 stabilizes protein expression and promotes surface retention of channel complexes.
Members of the K(+) channel-interacting protein (KChIP) family bind the distal N termini of members of the Shal subfamily of voltage-gated K(+) channel (Kv4) pore-forming (α) subunits to generate rapidly activating, rapidly inactivating neuronal A-type (I(A)) and cardiac transient outward (I(to)) currents. In heterologous cells, KChIP co-expression increases cell surface expression of Kv4 α subunits and Kv4 current densities, findings interpreted to suggest that Kv4·KChIP complex formation enhances forward trafficking of channels (from the endoplasmic reticulum or the Golgi complex) to the surface membrane. The results of experiments here, however, demonstrate that KChIP2 increases cell surface Kv4.2 protein expression (∼40-fold) by an order of magnitude more than the increase in total protein (∼2-fold) or in current densities (∼3-fold), suggesting that mechanisms at the cell surface regulate the functional expression of Kv4.2 channels. Additional experiments demonstrated that KChIP2 decreases the turnover rate of cell surface Kv4.2 protein by inhibiting endocytosis and/or promoting recycling. Unexpectedly, the experiments here also revealed that Kv4.2·KChIP2 complex formation stabilizes not only (total and cell surface) Kv4.2 but also KChIP2 protein expression. This reciprocal protein stabilization and Kv4·KChIP2 complex formation are lost with deletion of the distal (10 amino acids) Kv4.2 N terminus. Taken together, these observations demonstrate that KChIP2 differentially regulates total and cell surface Kv4.2 protein expression and Kv4 current densities.


Seasonal alterations in park visitation, amenity use, and physical activity--Grand Forks, North Dakota, 2012-2013.
Park amenities promote visitation and physical activity during summer. Physical activity declines during winter. Identifying park amenities that promote visitation during winter would increase year-round activity. The purpose of this study was to determine how park visitation, amenity choice, and physical activity intensity change across seasons. Physical activity intensity of children and adults was assessed at 16 parks in Grand Forks, North Dakota, during summer and fall of 2012, and winter and spring of 2013. Park visitation was highest in spring and lowest in winter. Amenity use varied by season. Parks with water splash pads were visited more during summer, and playgrounds and open spaces were visited more during spring. Ice rinks were visited most in winter. Physical activity intensity was lowest in summer and highest in winter for each age group. The activity intensity observed for all young age groups ranged from 2.7 to 2.9 metabolic equivalents in summer and greater than 3 metabolic equivalents in all other seasons. Adults' mean activity intensity was greater than 3 metabolic equivalents in winter. Information on park visitation, amenity use, and activity intensity across seasons is valuable; it can be used when designing or redesigning parks in order to promote year-round physical activity. Redesigning parks in cold climates to include ice rinks, sledding hills, cross-country skiing, and indoor areas for physical activity would increase winter visitation and allow the park to serve as a year-round resource for physical activity.


Injuries of the knee associated with fractures of the tibial shaft. Detection by examination under anesthesia: a prospective study.
Ligamentous injuries of the ipsilateral knee have been documented in association with 25 to 35 percent of fractures of the femoral shaft. However, to our knowledge, a similar association between ligamentous injuries of the knee and fracture of the tibial shaft has not been studied prospectively. To determine if there was such an association, a prospective study was conducted in which the knees of fifty patients who had a fracture of the ipsilateral tibial shaft of varying severity were examined manually while the patient was under general anesthesia. Eleven patients (22 percent) were found to have sustained an injury to at least one ligament of the knee that resulted in increased laxity of 2+ or more. One knee had dislocated. On the basis of the results of this study, we believe that, after stabilization of a fracture of the tibial shaft, it is essential to examine the knee thoroughly to identify any associated ligamentous injuries.


Cost minimization analysis of two treatment regimens for low-risk rhabdomyosarcoma in children: a report from the Children's Oncology Group.
Recent Children's Oncology Group trials for low-risk rhabdomyosarcoma attempted to reduce therapy while maintaining excellent outcomes. D9602 delivered 45 weeks of outpatient vincristine and dactinomycin (VA) for patients in Subgroup A. ARST0331 reduced the duration of therapy to 22 weeks but added four doses of cyclophosphamide to VA for patients in Subset 1. Failure-free survival was similar. We undertook a cost minimization comparison to help guide future decision-making. Addressing the costs of treatment from the healthcare perspective we modeled a simple decision-analytic model from aggregate clinical trial data. Medical care inputs and probabilities were estimated from trial reports and focused chart review. Costs of radiation, surgery and off-therapy surveillance were excluded. Unit costs were obtained from literature and national reimbursement and inpatient utilization databases and converted to 2012 US dollars. Model uncertainty was assessed with first-order sensitivity analysis. Direct medical costs were $46,393 for D9602 and $43,261 for ARST0331 respectively, making ARST0331 the less costly strategy. Dactinomycin contributed the most to D9602 total costs but varied with age (42-69%). Chemotherapy administration costs accounted for the largest proportion of ARST0331 total costs (39-57%). ARST0331 incurred fewer costs than D9602 under most alternative distributive models and alternative clinical practice assumptions. Cost analysis suggests that ARST0331 may incur fewer costs than D9602 from the healthcare system's perspective. Attention to the services driving the costs provides directions for future efficiency improvements. Future studies should prospectively consider the patient and family's perspective.


30-Day Readmission Rate and its Causes in General Surgical Patients.
To find out the 30-day readmission rate for different surgical procedures and determine the causes for readmissions. An audit report. Department of Surgery, The Aga Khan University Hospital, Karachi, in the calendar year 2014. Records were retrospectively reviewed for 30-day readmission rate and their cause in general surgical patients who underwent a general surgical procedure at The Aga Khan University Hospital, Karachi, in the year 2014. Out of 217 (6.4%) readmissions in 3,387 patients, more than 50% unplanned readmissions were avoidable. The highest readmission rate was after placement of feeding tubes (28.5%). More than half of the readmissions could have been prevented by proper patient/attendant education, good communication and provision of nursing services to these patients beyond the hospital.


Calreticulin functions as a molecular chaperone in the biosynthesis of myeloperoxidase.
Myeloperoxidase (MPO), a lysosomal heme protein found exclusively in neutrophils and monocytes, is necessary for efficient oxygen-dependent microbicidal activity. Acquisition of heme by the heme-free MPO precursor apopro-MPO appears to be a prerequisite for its subsequent proteolytic processing and advancement along the biosynthetic pathway to mature MPO. We present data indicating that calreticulin (CRT), a high capacity calcium-binding protein residing in the lumen of the endoplasmic reticulum of a wide variety of cells, interacts specifically with fully glycosylated apopro-MPO. Biosynthetically radiolabeled CRT (60 kDa) and apopro-MPO (90 kDa) were coprecipitated from PLB 985 cells by monospecific antiserum against CRT when the immunoprecipitations were performed either under nondenaturing conditions or following reversible crosslinking. Nonglycosylated MPO precursors synthesized in the presence of tunicamycin did not interact with CRT. The CRT-apopro-MPO interaction was restricted to an early phase of MPO biosynthesis, and CRT did not interact with the later appearing, heme-containing species of MPO, i.e. pro-MPO or the heavy subunit of mature MPO. These data show that CRT participates in the post-translational processing of MPO, perhaps by maintaining apopro-MPO in a conformation competent to accommodate insertion of the heme group. In this general way, CRT shares certain functional properties with the structurally homologous transmembrane calcium-binding endoplasmic reticulum protein calnexin. Both interact with glycosylated biosynthetic precursors of proteins selectively expressed in specialized cells.


Correlation between secretion and phospholipase D activation in differentiated HL60 cells.
Receptor-directed agonists including N-formylmethionyl-leucyl-phenylalanine (fMetLeuPhe), C5a, ATP and UTP all activate phospholipase D (PLD), which is accompanied by secretion in differentiated HL60 cells. Interference in the production of phosphatidase (PA) by the PLD pathway by diverting it towards the production of phosphatidylethanol (PEt) in the presence of ethanol leads to near-total inhibition of the secretion evoked by ATP and UTP and a partial inhibition of that evoked by fMetLeuPhe and C5a. In streptolysin-O-permeabilized cells, fMetLeuPhe is able to activate PLD, and this is dependent on the presence of a low concentration of guanosine 5'-[gamma-thio]-triphosphate (GTP[S]). Ca2+ (10 microM) and GTP[S] individually or in combination are also able to activate PLD and secretion. The stimulation of secretion in permeabilized cells stimulated by Ca2+ alone or fMetLeuPhe or GTP[S] is also abrogated when the production of PA is diverted to PEt by the presence of ethanol. Activation of PLD by GTP[S] or fMetLeuPhe is decreased if the cells are permeabilized first and GTP[S] or fMetLeuPhe is added subsequently. This corresponds well with the loss of the secretory response. We conclude that the ability of GTP[S] or fMetLeuPhe to stimulate secretion from permeabilized cells is dependent on a prior activation of the PLD signalling pathway. PA, generated as a consequence of PLD activation, acts as second messenger that can provide an initiating signal for secretion and is not required for exocytosis itself.


Two dimensional, electronic particle tracking in liquids with a graphene-based magnetic sensor array.
The investigation and control of liquid flow at the nanometer scale is a key area of applied research with high relevance to physics, chemistry, and biology. We introduce a method and a device that allows the spatial resolution of liquid flow by integrating an array of graphene-based magnetic (Hall) sensors that is used for tracking the movement of magnetic nanoparticles immersed in a liquid under investigation. With a novel device concept based on standard integration processes and experimentally verified material parameters, we numerically simulate the performance of a single sensor pixel, as well as the whole sensor array, for tracking magnetic nanoparticles having typical properties. The results demonstrate that the device enables (a) the detection of individual nanoparticles in the liquid with high accuracy and (b) the reconstruction of a particle's flow-driven trajectory across the integrated sensor array with sub-pixel precision as a function of time, in what we call the "Magnetic nanoparticle velocimetry" technique. Since the method does not rely on optical detection, potential lab-on-chip applications include particle tracking and flow analysis in opaque media at the sub-micron scale.


Induction therapy for adults with acute lymphoblastic leukemia: results of more than 1500 patients from the international ALL trial: MRC UKALL XII/ECOG E2993.
The international acute lymphoblastic leukemia (ALL) study was designed to prospectively define the optimal therapy for adults 60 years of age or younger with newly diagnosed ALL. All patients received identical induction therapy, and 91% achieved complete remission (CR). Patients 50 years of age or younger with a compatible sibling were assigned to undergo allogeneic transplantation; the others were randomly assigned to autologous transplantation or to consolidation/maintenance therapy for 2.5 years. Patients who did not achieve CR after induction had an overall survival rate of 5% compared with 45% for patients who achieved CR. Factors at diagnosis predictive of overall survival and disease-free survival were age (P = .001), white blood cell count less than 30 x 10(9)/L for B lineage or less than 100 x 10(9)/L for T lineage (P = .001) and immunophenotype, T lineage versus B lineage (P = .001). The data demonstrate that achieving CR with induction therapy is indispensable for long-term survival in adult patients with ALL. Furthermore, with a response rate greater than 90%, the induction regimen was highly efficacious as remission-inducing therapy. This large database has validated several previously identified independent prognostic factors in ALL, such as age, white blood cell count at presentation, cytogenetics, and immunophenotype. However, the achievement of CR within 4 weeks does not appear to be an independent prognostic factor.


Effect of metformin therapy on cardiac function and survival in a volume-overload model of heart failure in rats.
Advanced HF (heart failure) is associated with altered substrate metabolism. Whether modification of substrate use improves the course of HF remains unknown. The antihyperglycaemic drug MET (metformin) affects substrate metabolism, and its use might be associated with improved outcome in diabetic HF. The aim of the present study was to examine whether MET would improve cardiac function and survival also in non-diabetic HF. Volume-overload HF was induced in male Wistar rats by creating ACF (aortocaval fistula). Animals were randomized to placebo/MET (300 mg·kg(-1) of body weight·day(-1), 0.5% in food) groups and underwent assessment of metabolism, cardiovascular and mitochondrial functions (n=6-12/group) in advanced HF stage (week 21). A separate cohort served for survival analysis (n=10-90/group). The ACF group had marked cardiac hypertrophy, increased LVEDP (left ventricular end-diastolic pressure) and lung weight confirming decompensated HF, increased circulating NEFAs (non-esterified 'free' fatty acids), intra-abdominal fat depletion, lower glycogen synthesis in the skeletal muscle (diaphragm), lower myocardial triacylglycerol (triglyceride) content and attenuated myocardial (14)C-glucose and (14)C-palmitate oxidation, but preserved mitochondrial respiratory function, glucose tolerance and insulin sensitivity. MET therapy normalized serum NEFAs, decreased myocardial glucose oxidation, increased myocardial palmitate oxidation, but it had no effect on myocardial gene expression, AMPK (AMP-activated protein kinase) signalling, ATP level, mitochondrial respiration, cardiac morphology, function and long-term survival, despite reaching therapeutic serum levels (2.2±0.7 μg/ml). In conclusion, MET-induced enhancement of myocardial fatty acid oxidation had a neutral effect on cardiac function and survival. Recently reported cardioprotective effects of MET may not be universal to all forms of HF and may require AMPK activation or ATP depletion. No increase in mortality on MET supports its safe use in diabetic HF.


Homicide in discharged patients with schizophrenia and other psychoses: a national case-control study.
To investigate factors associated with homicide after discharge from hospital in patients with schizophrenia and other psychoses. All homicides committed by patients with psychosis within 6 months of hospital discharge were identified in Sweden from 1988-2001 and compared with patients with psychoses discharged over the same time period who did not subsequently commit any violent offences. Medical records were then collected, and data extracted using a validated protocol. Interrater reliability tests were performed on a subsample, and variables with poor reliability excluded from subsequent analyses. We identified 47 cases who committed a homicide within 6 months of discharge, and 105 controls who did not commit any violent offence after discharge. On univariate analyses, clinical factors on admission associated with homicide included evidence of poor self-care, substance misuse, and being previously hospitalized for a violent episode. Inpatient characteristics included having a severe mental illness for one year prior to admission. After-care factors associated with homicide were evidence of medication non-compliance and substance misuse. The predictive validity of combining two or three of these factors was not high. Depression appeared to be inversely associated with homicide, and there was no relationship with the presence of delusions or hallucinations. There are a number of potentially treatable factors that are associated with homicide in schizophrenia and other psychoses. Associations with substance misuse and treatment compliance could be the focus of therapeutic interventions if validated in other samples. However, their clinical utility in violence risk assessment remains uncertain.


Dynamic modulation of intrinsic functional connectivity by transcranial direct current stimulation.
Transcranial direct current stimulation (tDCS) is a noninvasive brain stimulation technique capable of modulating cortical excitability and thereby influencing behavior and learning. Recent evidence suggests that bilateral tDCS over both primary sensorimotor cortices (SM1) yields more prominent effects on motor performance in both healthy subjects and chronic stroke patients than unilateral tDCS over SM1. To better characterize the underlying neural mechanisms of this effect, we aimed to explore changes in resting-state functional connectivity during both stimulation types. In a randomized single-blind crossover design, 12 healthy subjects underwent functional magnetic resonance imaging at rest before, during, and after 20 min of unilateral, bilateral, and sham tDCS stimulation over SM1. Eigenvector centrality mapping (ECM) was used to investigate tDCS-induced changes in functional connectivity patterns across the whole brain. Uni- and bilateral tDCS over SM1 resulted in functional connectivity changes in widespread brain areas compared with sham stimulation both during and after stimulation. Whereas bilateral tDCS predominantly modulated changes in primary and secondary motor as well as prefrontal regions, unilateral tDCS affected prefrontal, parietal, and cerebellar areas. No direct effect was seen under the stimulating electrode in the unilateral condition. The time course of changes in functional connectivity in the respective brain areas was nonlinear and temporally dispersed. These findings provide evidence toward a network-based understanding regarding the underpinnings of specific tDCS interventions.


Pesticide Residues in Commercial Lettuce, Onion, and Potato Samples From Bolivia-A Threat to Public Health?
Bolivia does not have a surveillance program for pesticide residues in food. The few published studies have suggested that pesticide contamination in food may present a public health problem. Data are lacking for all foods except tomatoes and breast milk. In this study 10 potato, 10 onion, and 10 lettuce samples from La Paz were sampled on August 15, 2015 at a local market and screened for 283 pesticides. Residues of cypermethrin, chlorpyrifos, difenoconazol, or/and λ-cyhalothrin were detected in 50% of the lettuce samples, whereas no pesticides were found in potatoes and onions. In 20% of the lettuce samples, the measurements were above the maximum residue limits, and 2 or 3 pesticides were identified simultaneously. Washing almost halved the pesticide levels, but still 20% of the samples showed measurements above the limits. No samples contained concentrations of pesticides which alone or together would lead to exposures that exceeded the acceptable daily intake or the acute reference dose. To protect consumers from pesticide poisonings and chronic effects, the development of measures for prevention, control, and monitoring of food contamination by pesticides in Bolivia is suggested.


The effect of music therapy on hemodialysis patients' quality of life and depression symptoms.
The sources of stress involved in chronic kidney disease (CKD) can lead to depressive states, directly affecting the hemodialysis patient's quality of life (QOL). There are few reports and studies on therapeutic interventions that aim to minimize depressive symptoms in these patients and an even greater shortage of studies using music therapy. This study evaluated the effect of music therapy on QOL and depressive symptoms in hemodialysis patients. This was a music therapy intervention study in which 23 patients were evaluated regarding QOL and depression symptoms at two distinct stages - before and after the intervention. Eight sessions of music therapy were conducted, with two weekly sessions and an average duration of 75 minutes. The intervention was conducted by the music therapist herself, using specific music therapy techniques, besides voice and guitar to conduct harmonic and rhythmic support for the groups' sound-music production. The patients showed a significant reduction in depression symptoms (p < 0.001) and better QOL results, with significant differences in the following dimensions: functional capacity (p = 0.011), pain (p = 0.036), general health (p = 0.01), vitality (p = 0.004), mental health (p = 0.012), symptom and problem list (p = 0.01), and overall health (p = 0.01). Intervention with music therapy constitutes an effective option in the treatment and prevention of depressive symptoms and improved QOL of HD patients.


Design and demonstration of a colorless WDM-OFDMA PON system architecture achieving symmetric 20-Gb/s transmissions with residual interference compensation.
In this paper, we propose a two-tiered colorless WDM-OFDMA PON system architecture that draws strengths from each individual WDM and OFDM PON systems. Specifically, the two-tiered architecture enables a colorless transceiver to be shared by a group of ONUs, resulting in drastic reduction of the system cost. For achieving colorlessness via reusing downstream light sources, we discover the residual power of downstream signal unexpectedly springs back after transmissions, causing severe interference to the upstream signal, and thus limiting the data rate of the upstream signal. We devise a method of adopting a common dispersion compensation module at OLT to reduce the residual power over all wavelengths. Experimental results show that, with an improvement of upstream signal's SNR up to 10 dB, the system successfully achieves 20-Gb/s bidirectional OFDM-signal transmissions on the same wavelength over a 20-km SMF.


Effects of oxidation on the physicochemical properties of polyunsaturated lipid membranes.
The exposure of biological membranes to reactive oxygen species (ROS) plays an important role in many pathological conditions such as inflammation, infection, or sepsis. ROS also modulate signaling processes and produce markers for damaged tissue. Lipid peroxidation, mainly affecting polyunsaturated phospholipids, results in a complex mixture of oxidized products, which may dramatically alter membrane properties. Here, we have employed a set of biophysical and surface-chemical techniques, including neutron and X-ray scattering, to study the structural, compositional, and stability changes due to oxidative stress on phospholipid bilayers composed of lipids with different degrees of polyunsaturation. In doing so, we obtained real-time information about bilayer degradation under in situ UV exposure using neutron reflectometry. We present a set of interrelated physicochemical effects, including gradual increases in area per molecule, head group and acyl chain hydration, as well as bilayer thinning, lateral phase separation, and defect formation leading to content loss upon membrane oxidation. Such effects were observed to depend on the presence of polyunsaturated phospholipids in the lipid membrane, suggesting that these may also play a role in the complex oxidation processes occurring in cells.


A chemical view of the most ancient metazoa--biomarker chemotaxonomy of hexactinellid sponges.
Hexactinellid sponges are often considered to be the most ancient metazoans. Lipid biomarkers from 23 species were studied for information on their phylogenetic properties, particularly their disputed relation to the two other sponge classes (Demospongiae, Calcarea). The most prominent lipid compounds in the Hexactinellida comprise C28 to C32 polyenoic fatty acids. Their structures parallel the unique patterns found in demosponge membrane fatty acids ('demospongic acids') and strongly support a close phylogenetic association of the Demospongiae and the Hexactinellida. Both taxa also show unusual mid-chain methylated fatty acids (C15-C25) and irregular C25- and C40-isoprenoid hydrocarbons, tracers for specific eubacteria and Archaea, respectively. These biomarkers indicate a similar, highly conservative symbiont community, although some shift in the abundance of the associated microbiota was observed. The lack of these features in calcareous sponges further contradicts the still common view that Calcarea and Demospongiae are more closely related to each other than either is to the Hexactinellida.


Energy and nutrient intakes of Swedish children in relation to consumption of and habits associated with school lunch.
School lunches are provided free in Sweden, although some children choose not to eat school lunch. The aim of this study was to analyse Swedish children's total energy and nutrient intakes on weekdays by the frequency of school lunch consumption and to analyse energy and nutrient intakes from school lunches by sex. Factors associated with children's school lunch habits were also studied. Children in grades 2 and 5 ( n=1905) completed a food diary (school lunch data available for 1840 children) and the mean energy and nutrient intakes per day and per school lunch were calculated. The children also completed questions on the frequency of school lunch consumption and school lunch habits. Logistic regression was used to assess factors associated with school lunch habits. Children who reported eating school lunch every day had significantly higher energy and absolute nutrient intakes than children reporting eating school lunch less than five times a week, but not standardized for energy. Boys had significantly higher energy and absolute nutrient intakes from school lunches than girls, but not standardized for energy. Younger children and children who liked school lunches had higher odds of eating school lunch every day. Children in grade 5, those with a foreign background and those disliking school lunches had higher odds of omitting the main lunch component. Regular school lunch consumption was associated with a higher total intake for most nutrients, but not a better nutrient density. School lunch habits were associated with age, ethnic background and liking school lunches.


Human natural killer (NK) alloreactivity and its association with the major histocompatibility complex: ancestral haplotypes encode particular NK-defined haplotypes.
As ancestral haplotypes of the major histocompatibility complex (MHC) appear to define identical MHC haplotypes in unrelated individuals, unrelated individuals sharing the same ancestral haplotype should also share the same NK-defined allospecificities that have recently been shown to map to the human MHC. To test this prediction, multiple cell lines from unrelated individuals sharing the same ancestral haplotypes were tested for the NK-defined allospecificities. It was found that cells sharing the same ancestral haplotypes do have the same NK-defined specificities. Furthermore, the NK-defined phenotype of cells that possess two different ancestral haplotypes can be predicted from the NK-defined phenotypes of unrelated cells that are homozygous for the ancestral haplotypes concerned. Although the group 1 and 2 NK-defined allospecificities can be explained to some extent by HLA-C alleles, evidence is presented that additional genes may modify the phenotype conferred by HLA-C.


IDPicker 2.0: Improved protein assembly with high discrimination peptide identification filtering.
Tandem mass spectrometry-based shotgun proteomics has become a widespread technology for analyzing complex protein mixtures. A number of database searching algorithms have been developed to assign peptide sequences to tandem mass spectra. Assembling the peptide identifications to proteins, however, is a challenging issue because many peptides are shared among multiple proteins. IDPicker is an open-source protein assembly tool that derives a minimum protein list from peptide identifications filtered to a specified False Discovery Rate. Here, we update IDPicker to increase confident peptide identifications by combining multiple scores produced by database search tools. By segregating peptide identifications for thresholding using both the precursor charge state and the number of tryptic termini, IDPicker retrieves more peptides for protein assembly. The new version is more robust against false positive proteins, especially in searches using multispecies databases, by requiring additional novel peptides in the parsimony process. IDPicker has been designed for incorporation in many identification workflows by the addition of a graphical user interface and the ability to read identifications from the pepXML format. These advances position IDPicker for high peptide discrimination and reliable protein assembly in large-scale proteomics studies. The source code and binaries for the latest version of IDPicker are available from http://fenchurch.mc.vanderbilt.edu/ .


BRAF-V600E expression in precursor versus differentiated dendritic cells defines clinically distinct LCH risk groups.
Langerhans cell histiocytosis (LCH) is a clonal disorder with elusive etiology, characterized by the accumulation of CD207(+) dendritic cells (DCs) in inflammatory lesions. Recurrent BRAF-V600E mutations have been reported in LCH. In this study, lesions from 100 patients were genotyped, and 64% carried the BRAF-V600E mutation within infiltrating CD207(+) DCs. BRAF-V600E expression in tissue DCs did not define specific clinical risk groups but was associated with increased risk of recurrence. Strikingly, we found that patients with active, high-risk LCH also carried BRAF-V600E in circulating CD11c(+) and CD14(+) fractions and in bone marrow (BM) CD34(+) hematopoietic cell progenitors, whereas the mutation was restricted to lesional CD207(+) DC in low-risk LCH patients. Importantly, BRAF-V600E expression in DCs was sufficient to drive LCH-like disease in mice. Consistent with our findings in humans, expression of BRAF-V600E in BM DC progenitors recapitulated many features of the human high-risk LCH, whereas BRAF-V600E expression in differentiated DCs more closely resembled low-risk LCH. We therefore propose classification of LCH as a myeloid neoplasia and hypothesize that high-risk LCH arises from somatic mutation of a hematopoietic progenitor, whereas low-risk disease arises from somatic mutation of tissue-restricted precursor DCs.


NLRC5 knockdown in chicken macrophages alters response to LPS and poly (I:C) stimulation.
NLRC5 is a member of the CARD domain containing, nucleotide-binding oligomerization (NOD)-like receptor (NLR) family, which recognizes pathogen-associated molecular patterns (PAMPs) and initiates an innate immune response leading to inflammation and/or cell death. However, the specific role of NLRC5 as a modulator of the inflammatory immune response remains controversial. It has been reported to be a mediator of type I IFNs, NF-kB, and MHC class I gene. But no study on NLRC5 function has been reported to date in chickens. In the current study, we investigated the role of NLRC5 in the regulation of IFNA, IFNB, IL-6, and MHC class I in the chicken HD11 macrophage cell line, by using RNAi technology. HD11 cells were transfected with one of five siRNAs (s1, s2, s3, negative-siRNA, or a mixture of s1, s2, s3-siRNAs). After 24 hours, cells were exposed to LPS or poly (I:C) or a vehicle control. Gene expression of NLRC5, IFNA, IFNB, IL-6, and MHC class I at 2, 4, 6, and 8 hours post stimulation (hps) was quantified by qPCR. The expression of NLRC5, IFNA, IFNB, and IL-6 genes in negative irrelevant transfection controls was up-regulated at 2 hps after LPS treatment compared to the vehicle controls. S3-siRNA effectively knocked down NLRC5 expression at 4 hps, and the expression of IFNA and IFNB (but not IL-6 and MHC class I) was also down-regulated at 4 hps in s3-siRNA transfected cells, compared to negative irrelevant transfection controls. Stimulation by LPS appeared to relatively restore the decrease in NLRC5, IFNA, and IFNB expression, but the difference is not significant. Functional characterization of chicken NLRC5 in an in vitro system demonstrated its importance in regulating intracellular molecules involved in inflammatory response. The knockdown of NLRC5 expression negatively mediates gene expression of IFNA and IFNB in the chicken HD11 cell line; therefore, NLRC5 likely has a role in positive regulation of IFNA and IFNB expression. No direct relationship was found between NLRC5 knockdown and IL-6 and MHC class I expression. Future studies will further clarify the roles of NLRC5 and other NLRs in infectious diseases of chickens and may increase the efficacy of antiviral vaccine design.


Susceptibility of Baltic and East Atlantic salmon Salmo salar stocks to Gyrodactylus salaris (Monogenea).
The susceptibility of a Baltic salmon stock Salmo salar (Indalsälv, central Sweden) to Norwegian Gyrodactylus salaris (Figga strain, central Norway) was experimentally tested and compared with previously obtained results on East Atlantic salmon (Lierelva, SE Norway). Contrary to expectation, the Baltic salmon, which had no prior exposure to this parasite strain, appeared almost as susceptible as the Norwegian salmon parr that naturally experience G. salaris-induced mortality. Individually isolated salmon of both stocks sustained G. salaris infections with little evidence of innate resistance. A few individuals of the Indalsälv stock controlled their infection from the beginning, but overall there was considerable heterogeneity in the course of infection in both stocks. On individual hosts, G. salaris growth rates declined steadily throughout the infection, a trend which was particularly marked amongst the Lierelva stock. On shoaling Lierelva fish, there was some evidence of reduced parasite population growth towards the end of the infection; this was not apparent in Indalsälv fishes. These results reflect a growing awareness that not all Baltic salmon may be resistant to Norwegian G. salaris, and that Norwegian and Baltic G. salaris strains may differ in virulence. Consequently, management decisions concerning this parasite-host system should be based upon the actual, and tested, susceptibility of stocks under consideration and not upon identification of stocks as either Atlantic or Baltic.


Characterization of a full-length infectious cDNA clone and a GFP reporter derivative of the oncolytic picornavirus SVV-001.
Seneca Valley virus (SVV-001) is an oncolytic picornavirus with selective tropism for a subset of human cancers with neuroendocrine differentiation. To characterize further the specificity of SVV-001 and its patterns and kinetics of intratumoral spread, bacterial plasmids encoding a cDNA clone of the full-length wild-type virus and a derivative virus expressing GFP were generated. The full-length cDNA of the SVV-001 RNA genome was cloned into a bacterial plasmid under the control of the T7 core promoter sequence to create an infectious cDNA clone, pNTX-09. A GFP reporter virus cDNA clone, pNTX-11, was then generated by cloning a fusion protein of GFP and the 2A protein from foot-and-mouth disease virus immediately following the native SVV-001 2A sequence. Recombinant GFP-expressing reporter virus, SVV-GFP, was rescued from cells transfected with in vitro RNA transcripts from pNTX-11 and propagated in cell culture. The proliferation kinetics of SVV-001 and SVV-GFP were indistinguishable. The SVV-GFP reporter virus was used to determine that a subpopulation of permissive cells is present in small-cell lung cancer cell lines previously thought to lack permissivity to SVV-001. Finally, it was shown that SVV-GFP administered to tumour-bearing animals homes in to and infects tumours whilst having no detectable tropism for normal mouse tissues at 1×10(11) viral particles kg(-1), a dose equivalent to that administered in ongoing clinical trials. These infectious clones will be of substantial value in further characterizing the biology of this virus and as a backbone for the generation of additional oncolytic derivatives.


Recruitment of neutrophils during IgE-dependent cutaneous late phase reactions in the mouse is mast cell-dependent. Partial inhibition of the reaction with antiserum against tumor necrosis factor-alpha.
Much of the clinically important pathology associated with IgE-dependent disorders is thought to reflect the actions of the blood-borne leukocytes recruited during these responses. To evaluate the extent to which mast cells are responsible for the leukocyte infiltration associated with IgE-dependent cutaneous reactions, we attempted to elicit these responses in normal mice, genetically mast cell-deficient W/Wv mice, and in W/Wv mice selectively repaired of their mast cell deficiency by the intradermal injection of cultured mast cells derived from the congenic normal (+/+) mice. We found that the tissue swelling associated with IgE-dependent passive cutaneous anaphylaxis reactions developed rapidly and diminished markedly from 2 to 4 h after antigen challenge, but remained detectable for at least 24 h after elicitation of the responses. Infiltration of leukocytes (predominantly neutrophils) also occurred at these sites, but reached maximal levels 6-12 h after antigen challenge, persisted at high levels for 24 h, and largely waned by 48 h. Virtually all of the tissue swelling and leukocyte infiltration associated with IgE-dependent cutaneous reactions was mast cell dependent. Intradermal injection of 40 U of recombinant murine TNF-alpha (rmTNF-alpha) elicited neutrophil infiltration similar in magnitude and kinetics to that observed after IgE-dependent mast cell degranulation. A rabbit anti-rmTNF-alpha (R anti-rmTNF-alpha) antiserum, which was able to inhibit 84% of the neutrophil infiltration observed after i.d. injection of rmTNF-alpha, inhibited IgE-, and mast cell-dependent leukocyte infiltration by 47 +/- 7% in three separate experiments. These findings indicate that TNF-alpha contributes to mast cell-dependent recruitment of leukocytes during IgE-dependent cutaneous late phase reactions, but suggest that other mast cell-associated mediators probably also contribute to this response.


American Academy of Pediatrics. Clinical report--sport-related concussion in children and adolescents.
Sport-related concussion is a "hot topic" in the media and in medicine. It is a common injury that is likely underreported by pediatric and adolescent athletes. Football has the highest incidence of concussion, but girls have higher concussion rates than boys do in similar sports. A clear understanding of the definition, signs, and symptoms of concussion is necessary to recognize it and rule out more severe intracranial injury. Concussion can cause symptoms that interfere with school, social and family relationships, and participation in sports. Recognition and education are paramount, because although proper equipment, sport technique, and adherence to rules of the sport may decrease the incidence or severity of concussions, nothing has been shown to prevent them. Appropriate management is essential for reducing the risk of long-term symptoms and complications. Cognitive and physical rest is the mainstay of management after diagnosis, and neuropsychological testing is a helpful tool in the management of concussion. Return to sport should be accomplished by using a progressive exercise program while evaluating for any return of signs or symptoms. This report serves as a basis for understanding the diagnosis and management of concussion in children and adolescent athletes.


A primary care, multi-disciplinary disease management program for opioid-treated patients with chronic non-cancer pain and a high burden of psychiatric comorbidity.
Chronic non-cancer pain is a common problem that is often accompanied by psychiatric comorbidity and disability. The effectiveness of a multi-disciplinary pain management program was tested in a 3 month before and after trial. Providers in an academic general medicine clinic referred patients with chronic non-cancer pain for participation in a program that combined the skills of internists, clinical pharmacists, and a psychiatrist. Patients were either receiving opioids or being considered for opioid therapy. The intervention consisted of structured clinical assessments, monthly follow-up, pain contracts, medication titration, and psychiatric consultation. Pain, mood, and function were assessed at baseline and 3 months using the Brief Pain Inventory (BPI), the Center for Epidemiological Studies-Depression Scale scale (CESD) and the Pain Disability Index (PDI). Patients were monitored for substance misuse. Eighty-five patients were enrolled. Mean age was 51 years, 60% were male, 78% were Caucasian, and 93% were receiving opioids. Baseline average pain was 6.5 on an 11 point scale. The average CESD score was 24.0, and the mean PDI score was 47.0. Sixty-three patients (73%) completed 3 month follow-up. Fifteen withdrew from the program after identification of substance misuse. Among those completing 3 month follow-up, the average pain score improved to 5.5 (p = 0.003). The mean PDI score improved to 39.3 (p < 0.001). Mean CESD score was reduced to 18.0 (p < 0.001), and the proportion of depressed patients fell from 79% to 54% (p = 0.003). Substance misuse was identified in 27 patients (32%). A primary care disease management program improved pain, depression, and disability scores over three months in a cohort of opioid-treated patients with chronic non-cancer pain. Substance misuse and depression were common, and many patients who had substance misuse identified left the program when they were no longer prescribed opioids. Effective care of patients with chronic pain should include rigorous assessment and treatment of these comorbid disorders and intensive efforts to insure follow up.


Distinct cortical anatomy linked to subregions of the medial temporal lobe revealed by intrinsic functional connectivity.
The hippocampus and adjacent cortical structures in the medial temporal lobe (MTL) contribute to memory through interactions with distributed brain areas. Studies of monkey and rodent anatomy suggest that parallel pathways converge on distinct subregions of the MTL. To explore the cortical areas linked to subregions of the MTL in humans, we examined cortico-cortical and hippocampal-cortical correlations using high-resolution, functional connectivity analysis in 100 individuals. MTL seed regions extended along the anterior to posterior axis and included hippocampus and adjacent structures. Results revealed two separate brain pathways that correlated with distinct subregions within the MTL. The body of the hippocampus and posterior parahippocampal cortex correlated with lateral parietal cortex, regions along the posterior midline including posterior cingulate and retrosplenial cortex, and ventral medial prefrontal cortex. By contrast, anterior hippocampus and the perirhinal/entorhinal cortices correlated with distinct regions in the lateral temporal cortex extending into the temporal pole. The present results are largely consistent with known connectivity in the monkey and provide a novel task-independent dissociation of the parallel pathways supporting the MTL memory system in humans. The cortical pathways include regions that have undergone considerable areal expansion in humans, providing insight into how the MTL memory system has evolved to support a diverse array of cognitive domains.


Osteoarthritis and telomere shortening.
Osteoarthritis is the most common disease of joints caused by degradation of articular cartilage and subchondral bone. It is classified as primary form with unknown cause and as secondary form with known etiology. Genetic and epigenetic factors interact with environmental factors and contribute to the development of primary osteoarthritis. Thus far, many polymorphisms associated with osteoarthritis have been identified and recent studies also indicate the involvement of epigenetic factors (e.g., telomere shortening) in the initiation of this disorder. Accelerated shortening of telomeres was detected in osteoarthritis and other age-related diseases. Studies revealed that telomere length is severely reduced in blood leukocytes and chondrocytes of patients with osteoarthritis, and this may contribute to the initiation and development of osteoarthritis, whose major cause is still unknown.


Rheumatoid arthritis antirheumatic drug trials. III. Setting the delta for clinical trials of antirheumatic drugs--results of a consensus development (Delphi) exercise.
Defining the minimum clinically important difference or delta to be detected in a clinical trial depends on a number of factors including the research hypothesis, patient characteristics, the nature of the intervention and the trial design. In 2 previous studies, we have developed standardized procedures for conducting outcome measurement based on current Food and Drug Administration and European League Against Rheumatism guidelines for RA clinical trials, and thereafter, determined the standard deviation for these outcome measures. In the final component of this series of studies, we employed a Delphi technique to establish estimates for delta and calculated the sample size requirements under different conditions of Type I and Type II error probabilities.


Retrospective Assessment of Occupational Exposures for the GENEVA Study of ALS among Military Veterans.
This paper describes the retrospective exposure assessment conducted to assess occupational exposures for the Genes and Environmental Exposures in Veterans (GENEVA) study, a case-control study investigating the joint contribution of genetics and environmental exposures to the risk of amyotrophic lateral sclerosis (ALS) among military veterans. Occupational histories for 1597 study participants collected as part of the GENEVA study were the basis for this retrospective exposure assessment. The data set included 15528 jobs held from 1924 to 2010, representing 4539 unique industry and occupation (I&O) combinations. Three industrial hygiene experts were recruited to independently rate occupational exposures to specific agents previously associated with an increased risk of ALS. Utilizing information on industry, job title, tasks performed, and materials used for each job held, raters assigned exposures associated with each I&O for the 'current time' defined as the period after 1995 (post-1995). The exposure assessment targeted agents identified as potential occupational risk factors for ALS. Experts rated semi-quantitatively exposure intensity in five exposure categories (0-4) for Group A agents (lead, formaldehyde, hydrocarbon solvents, and chlorinated solvents) and qualitatively as yes/no (1/0) exposed for Group B agents (mercury, selenium, arsenic, polychlorinated biphenyls, electromagnetic field, pesticides, and viral agents). Confidence scores (0-3) were reported for every I&O rated based on raters' experience with that industry and/or job. Each I&O was assigned an average exposure score of the raters and an alternative exposure rating was developed for each I&O by excluding low confidence (<2) scores before averaging. Exposure reconstruction for jobs held pre-1995 was done by comparing exposure data extracted from the OSHA Chemical Exposure and Health Database (CEHD) during pre-1995 and post-1995. For agents with limited exposure data in the CEHD, pre-1995 exposures were determined based on raters' judgment. The proportion of I&O combinations determined to be 'exposed' ranged from 0.1 to 26% across different agents, with the highest values corresponding to hydrocarbon solvents and the lowest to selenium. Industries with the highest proportion of exposed records include manufacturing, utilities, healthcare, and military with non-combat jobs. Analyses for raters' reliability showed the best agreement between the raters when rating exposure to viral agents (kappa = 0.67), hydrocarbon solvents (kappa = 0.53), and lead (kappa = 0.50). The proportion of 'exposed' I&O combinations increased for hydrocarbon solvents, chlorinated solvents, and pesticides when exposure ratings were adjusted by raters' confidence. Compared to post-1995, exposures in the earlier period (pre-1995) were deemed higher or the same for most of the agents and lower for formaldehyde and electromagnetic field exposures. Our results indicate that using raters' confidence assessment in determining exposure scores increases both the proportion of I&O combinations regarded as exposed and the intensity scores, suggesting raters tend to be conservative in their assessment when they lack detailed knowledge of an industry or job.


Assessing point-of-use ultraviolet disinfection for safe water in urban developing communities.
Residents of urban developing communities often have a tap in their home providing treated and sometimes filtered water but its microbial quality cannot be guaranteed. Point-of-use (POU) disinfection systems can provide safe drinking water to the millions who lack access to clean water in urban communities. While many POU systems exist, there are several concerns that can lead to low user acceptability, including low flow rate, taste and odor issues, high cost, recontamination, and ineffectiveness at treating common pathogens. An ultraviolet (UV) POU system was constructed utilizing developing community-appropriate materials and simple construction techniques based around an inexpensive low-wattage, low pressure UV bulb. The system was tested at the bench scale to characterize its hydrodynamic properties and microbial disinfection efficacy. Hydraulically the system most closely resembled a plug flow reactor with minor short-circuiting. The system was challenge tested and validated for a UV fluence of 50 mJ/cm(2) and greater, over varying flow rates and UV transmittances, corresponding to a greater than 4 log reduction of most pathogenic bacteria, viruses, and protozoa of public health concern. This study presents the designed system and testing results to demonstrate the potential architecture of a low-cost, open-source UV system for further prototyping and field-testing.


Vasopressin-mediated contractions in mesenteric arteries from spontaneously hypertensive rats: differences in response compared with Wistar-Kyoto animals.
1. The sensitivity of mesenteric resistance arterioles to [arginine]vasopressin (AVP) was investigated in spontaneously hypertensive rats (SHR) and control Wistar-Kyoto rats (WKY). No difference in pAVP (-log dose of AVP producing 50% of the maximum response) was observed [SHR 0.21 +/- 0.03 m-units/ml (n = 10) vs WKY 0.15 +/- 0.06 m-units/ml (n = 9)], although SHR vessels exhibited greater absolute tension development. 2. Both strains of rat displayed tachyphylaxis to repeated stimulation with AVP, and oscillatory tension changes were observed in all vessels from SHR and rarely in WKY vessels at activating concentrations of AVP. 3. AVP did not elicit a contractile response after noradrenaline-induced calcium depletion. 4. After vessels were depleted of calcium by using a combination of calcium-free media and noradrenaline stimulation, restoration of calcium in the presence of AVP elicited a greater contractile response in SHR vessels. 5. The results therefore provide evidence for an increased calcium response to AVP in SHR resistance vessels, although this was only demonstrable by calcium recovery experiments.


Wolf in Sheep's Clothing: Clostridioides difficile Biofilm as a Reservoir for Recurrent Infections.
The microbiota inhabiting the intestinal tract provide several critical functions to its host. Microorganisms found at the mucosal layer form organized three-dimensional structures which are considered to be biofilms. Their development and functions are influenced by host factors, host-microbe interactions, and microbe-microbe interactions. These structures can dictate the health of their host by strengthening the natural defenses of the gut epithelium or cause disease by exacerbating underlying conditions. Biofilm communities can also block the establishment of pathogens and prevent infectious diseases. Although these biofilms are important for colonization resistance, new data provide evidence that gut biofilms can act as a reservoir for pathogens such as Clostridioides difficile. In this review, we will look at the biofilms of the intestinal tract, their contribution to health and disease, and the factors influencing their formation. We will then focus on the factors contributing to biofilm formation in C. difficile, how these biofilms are formed, and their properties. In the last section, we will look at how the gut microbiota and the gut biofilm influence C. difficile biofilm formation, persistence, and transmission.


Intravenous administration of prochlorperazine by 15-minute infusion versus 2-minute bolus does not affect the incidence of akathisia: a prospective, randomized, controlled trial.
We sought to compare the rate of akathisia after administration of intravenous prochlorperazine as a 2-minute bolus or 15-minute infusion. We conducted a prospective, randomized, double-blind study in the emergency department of a central-city teaching hospital. Patients aged 18 years or older treated with prochlorperazine for headache, nausea, or vomiting were eligible for inclusion. Study participants were randomized to receive 10 mg of prochlorperazine administered intravenously by means of 2-minute push (bolus group) or 10 mg diluted in 50 mL of normal saline solution administered by means of intravenous infusion during a 15-minute period (infusion group). The main outcome was the number of study participants experiencing akathisia within 60 minutes of administration. Akathisia was defined as either a spontaneous report of restlessness or agitation or a change of 2 or more in the patient-reported akathisia rating scale and a change of at least 1 in the investigator-observed akathisia rating scale. The intensity of headache and nausea was measured with a 100-mm visual analog scale. One hundred patients were enrolled. One study participant was excluded after protocol violation. Seventy-three percent (73/99) of the study participants were treated for headache and 70% (70/99) for nausea. In the bolus group, 26.0% (13/50) had akathisia compared with 32.7% (16/49) in the infusion group (Delta=-6.7%; 95% confidence interval [CI] -24.6% to 11.2%). The difference between the bolus and infusion groups in the percentage of participants who saw a 50% reduction in their headache intensity within 30 minutes was 11.8% (95% CI -9.6% to 33.3%). The difference in the percentage of patients with a 50% reduction in their nausea was 12.6% (95% CI -4.6% to 29.8%). A 50% reduction in the incidence of akathisia when prochlorperazine was administered by means of 15-minute intravenous infusion versus a 2-minute intravenous push was not detected. The efficacy of prochlorperazine in the treatment of headache and nausea likewise did not appear to be affected by the rate of administration, although no formal statistical comparisons were made.


The use of ecological footprint in estimating the Environmental Kuznets Curve hypothesis for BRICST by considering cross-section dependence and heterogeneity.
A vast body of literature estimates the impact of economic growth on environmental degradation in the framework of EKC model. Typical empirical studies proxy environmental degradation with CO2 emissions; however, this indicator does not consider the complex nature of environmental degradation. To fulfill this omission, ecological footprint that tracks the use of multiple categories of productive surface areas is used as proxy for the environment. Moreover, studies that do not consider issues of heterogeneity and cross-sectional dependence may not produce reliable outcomes. Hence, the present study re-investigates the validity of the EKC hypothesis for BRICST (Brazil, Russia, India, China, South Africa, Turkey) by using ecological footprint and considering the mentioned issues in the estimation process. Based on the annual data covering the period of 1980-2014, excluding Russia due to data unavailability, empirical results show that the EKC hypothesis is not valid, and energy intensity and energy structure are important determinants of environmental degradation. In line with the empirical outputs, possible policy suggestions are discussed in the present study.


Evaluation of implants coated with recombinant human bone morphogenetic protein-2 and vacuum-dried using the critical-size supraalveolar peri-implant defect model in dogs.
Endosseous implants coated with recombinant human bone morphogenetic protein-2 (rhBMP-2) in a laboratory bench setting and air-dried induce relevant bone formation but also resident bone remodeling. Thus, the objective of this study is to evaluate the effect of implants fully or partially coated with rhBMP-2 and vacuum-dried using an industrial process on local bone formation and resident bone remodeling. Twelve male adult Hound Labrador mongrel dogs were used. Critical-size, supraalveolar, peri-implant defects received titanium porous oxide surface implants coated in their most coronal aspect with rhBMP-2 (coronal-load, six animals), or by immersion of the entire implant in a rhBMP-2 solution (soak-load, six animals) for a total of 30 μg rhBMP-2 per implant. All implants were vacuum-dried. The animals were sacrificed at 8 weeks for histometric evaluation. Clinical healing was unremarkable. Bone formation was not significantly affected by the rhBMP-2 application protocol. New bone height and area averaged (± SE) 3.2 ± 0.5 versus 3.6 ± 0.3 mm, and 2.3 ± 0.5 versus 2.6 ± 0.8 mm(2) for coronal-load and soak-load implants, respectively (P >0.05). The corresponding bone density and bone-implant contact registrations averaged 46.7% ± 5.8% versus 31.6% ± 4.4%, and 28% ± 5.6% versus 36.9% ± 3.4% (P >0.05). In contrast, resident bone remodeling was significantly influenced by the rhBMP-2 application protocol. Peri-implant bone density averaged 72.2% ± 2.1% for coronal-load versus 60.6% ± 4.7% for soak-load implants (P <0.05); the corresponding bone-implant contact averaged 70.7% ± 6.1% versus 47.2% ± 6.0% (P <0.05). Local application of rhBMP-2 and vacuum-drying using industrial process seems to be a viable technology to manufacture implants that support local bone formation and osseointegration. Coronal-load implants obviate resident bone remodeling without compromising local bone formation.


Enzymatic regulation of calcium current in dialyzed and intact molluscan neurons.
Isolated neurons of Helix aspersa were dialyzed and voltage clamped under conditions that isolate the Ca current. The rapid time-dependent run-down, or washout, of Ca current could be slowed by addition of 1 mM EGTA to the dialysis solution. A more effective means of slowing washout was the use of agents that promote protein phosphorylation, such as cAMP, Mg-ATP and the catalytic subunit (CS) of cAMP-dependent protein kinase, along with leupeptin, a tripeptide inhibitor of proteases. In the presence of these agents, no internal EGTA was required to prevent Ca current washout. Thus, during dialysis with 100 microM leupeptin, 7 mM Mg-ATP and 20 micrograms/ml CS, the Ca current remained stable for up to several hours. The rate of Ca-dependent inactivation of the current that occurs during a depolarizing step showed only a small decline during prolonged dialysis. Under these conditions, introduction of 10 microM calmodulin plus 40 micrograms/ml calcineurin, a Ca-calmodulin-dependent phosphatase, caused a significant increase in the rate of Ca current inactivation during a depolarizing step. This increase in rate of inactivation, as well as the original inactivation, was eliminated by introduction of EGTA or replacement of external Ca with Ba, results that are consistent with the ion dependency for activation of calcineurin. When internal ATP was replaced with ATP-gamma-S, a hydrolysis-resistant analogue, the rate of Ca current inactivation slowed, providing further evidence that inactivation involves a dephosphorylation.(ABSTRACT TRUNCATED AT 250 WORDS)


Neurogenic and proneural genes control cell fate specification in the Drosophila endoderm.
The Drosophila endoderm segregates into three non-neural cell types, the principle midgut epithelial cells, the adult midgut precursors, and the interstitial cell precursors, early in development. We show that this process occurs in the absence of mesoderm and requires proneural and neurogenic genes. In neurogenic mutants the principle midgut epithelial cells are missing and the other two cell types develop in great excess. Consequently, the midgut epithelium does not form. In achaete-scute complex and daughterless mutants the interstitial cell precursors do not develop and the number of adult midgut precursors is strongly reduced. Development of the principle midgut epithelial cells and formation of the midgut epithelium is restored in neurogenic proneural double mutants. The neurogenic/proneural genes are, in contrast to the neuroectoderm, not expressed in small clusters of cells but initially homogeneously in the endoderm suggesting that no prepattern exists which determines the position of the segregating cells. Hence, the segregation pattern solely depends on neurogenic/proneural gene interaction. Proneural genes are required but not sufficient to determine specific cell fates because they are required for cell type specification in both ectoderm and endoderm. Our data also suggest that the neurogenic/proneural genes are involved in the choice between epithelial versus mesenchymal cell morphologies.


Salt-induced hypertension in the 'Sabra' rat strain: influence of nifedipine treatment.
The effects of chronic dietary salt-loading and nifedipine therapy on hypertension-prone (SBH), -resistant (SBN) and parental (SB) Sabra rats were investigated. Salt diet for 12 weeks resulted in a sustained hypertension and heart hypertrophy only in SBH. Nifedipine therapy (300 p.p.m. = 300 mg/kg of food) introduced after week 7 on a salt diet, achieved small changes in salt-loaded SBN and SB rats, but resulted in a marked decrease in blood pressure in SBH rats within 1 week and in a regression of cardiac hypertrophy. Plasma renin activity rose slightly in nifedipine treated SB and SBN rats, but decreased significantly in treated SBH rats. Histopathological investigations revealed hypertensive vasculopathy in three out of nine untreated SBH rats, whereas there were no morphological changes in the treated rats.


How Different are Men Who Do Not Know Their HIV Status from Those Who Do? Results from an U.S. Online Study of Gay and Bisexual Men.
We compared self-described HIV-positive (31.6 %, n = 445), HIV-negative (56.8 %, n = 801), and HIV-unknown (11.6 %, n = 164) gay and bisexual men on sociodemographic and behavioral characteristics. Participants from across the U.S. were enrolled via a popular sexual networking website to complete an online survey. In total, 44.8 % of HIV-negative and HIV-unknown men said they had not been tested for HIV in the CDC-recommended last 6 months. HIV-unknown men significantly differed from HIV-negative and HIV-positive men in sexual behavior and HIV status disclosure patterns. HIV-unknown men were more willing than HIV-negative men to take PrEP; however, HIV-unknown men were significantly less likely than others to have health insurance or a primary care provider. Given the observed differences, researchers should consider analyzing men who are HIV-unknown distinctly from HIV-negative and HIV-positive men.


Rethinking research ethics.
Contemporary research ethics policies started with reflection on the atrocities perpetrated upon concentration camp inmates by Nazi doctors. Apparently, as a consequence of that experience, the policies that now guide human subject research focus on the protection of human subjects by making informed consent the centerpiece of regulatory attention. I take the choice of context for policy design, the initial prioritization of informed consent, and several associated conceptual missteps, to have set research ethics off in the wrong direction. The aim of this paper is to sort out these confusions and their implications and to offer instead a straightforward framework for considering the ethical conduct of human subject research. In the course of this discussion I clarify different senses of autonomy that have been confounded and present more intelligible justifications for informed consent. I also take issue with several of the now accepted dogmas that govern research ethics. These include: the primacy of informed consent, the protection of the vulnerable, the substitution of beneficence for research's social purpose, and the introduction of an untenable distinction between innovation and research.


Quantification of heterosubtypic immunity between avian influenza subtypes H3N8 and H4N6 in multiple avian host species.
Low-pathogenicity avian influenza virus (LPAIV) can lead to epizootics that cause economic losses in poultry or the emergence of human-infectious strains. LPAIVs experience a complex immunity landscape as they are endemic in numerous host species, and many antigenically distinct strains co-circulate. Prevention and control of emergence of detrimental strains requires an understanding of infection/transmission characteristics of the various subtypes in different hosts, including interactions between subtypes. In order to develop analytical frameworks for examining control efficacy, quantification of heterosubtypic immunity interactions is fundamental. However, these data are scarce, especially for wild avian subtypes in natural hosts. Consequently, in this study, three host species (mallards, quail and pheasants) were infected with two LPAIV subtypes isolated from wild birds: H3N8 and H4N6. The recovered hosts were also reinfected with the alternate subtype to measure the effects of heterosubtypic immunity. Oropharyngeal and cloacal swabs were collected and viral RNA load was quantified by real-time RT-PCR. For secondary infections in recovered hosts, peak viral load was up to four orders of magnitude lower and shedding length was up to 4 days shorter. However, both the magnitude and presence of heterosubtypic immunity varied across specific host species/subtype combinations. Using a mathematical model of virus replication, the variation in virus replication dynamics due to host individuals was quantified. It was found that accounting for individual heterogeneity is important for drawing accurate conclusions about treatment effects. These results are relevant for developing epidemiological models to inform control practices and for analysing virus replication data.


Relationship between psychological stress and reproductive outcome in women undergoing in vitro fertilization treatment: psychological and neurohormonal assessment.
To evaluate whether psychological stress, as well as changes in hypothalamus-pituitary-adrenal (HPA) axis and sympathetic nervous system (SNS) at different time points during a first in vitro fertilization (IVF) cycle, correlates with the reproductive outcome. A prospective study was conducted in 264 women undergoing IVF or intracytoplasmic sperm injection (ICSI) treatment between January 2009 and March 2010. Standardized psychological questionnaires were used to assess anxiety and depression. Norepinephrine and cortisol in serum were measured with specific assays. The non-pregnant women reported higher anxiety and depression scores at the pregnancy detection day compared with the pregnant group. Lower levels of norepinephrine and cortisol at the time of oocyte retrieval and lower levels of cortisol at the time of pregnancy test were found in women with successful treatment. Significant increases in serum norepinephrine and cortisol values were observed during ovarian stimulation. State Anxiety scores were negatively correlated with live birth rate, and positively associated with serum norepinephrine and cortisol values. State anxiety is associated with both pregnancy rate and live birth rate in IVF patients, an effect that is partly mediated by activities in the HPA and SNS.


Ranibizumab for neovascular age-related macular degeneration.
Ranibizumab--a recombinant, humanized, monoclonal antibody Fab that neutralizes all active forms of vascular endothelial growth factor A--has been evaluated for the treatment of neovascular age-related macular degeneration. In this multicenter, 2-year, double-blind, sham-controlled study, we randomly assigned patients with age-related macular degeneration with either minimally classic or occult (with no classic lesions) choroidal neovascularization to receive 24 monthly intravitreal injections of ranibizumab (either 0.3 mg or 0.5 mg) or sham injections. The primary end point was the proportion of patients losing fewer than 15 letters from baseline visual acuity at 12 months. We enrolled 716 patients in the study. At 12 months, 94.5% of the group given 0.3 mg of ranibizumab and 94.6% of those given 0.5 mg lost fewer than 15 letters, as compared with 62.2% of patients receiving sham injections (P<0.001 for both comparisons). Visual acuity improved by 15 or more letters in 24.8% of the 0.3-mg group and 33.8% of the 0.5-mg group, as compared with 5.0% of the sham-injection group (P<0.001 for both doses). Mean increases in visual acuity were 6.5 letters in the 0.3-mg group and 7.2 letters in the 0.5-mg group, as compared with a decrease of 10.4 letters in the sham-injection group (P<0.001 for both comparisons). The benefit in visual acuity was maintained at 24 months. During 24 months, presumed endophthalmitis was identified in five patients (1.0%) and serious uveitis in six patients (1.3%) given ranibizumab. Intravitreal administration of ranibizumab for 2 years prevented vision loss and improved mean visual acuity, with low rates of serious adverse events, in patients with minimally classic or occult (with no classic lesions) choroidal neovascularization secondary to age-related macular degeneration. (ClinicalTrials.gov number, NCT00056836 [ClinicalTrials.gov].).


Fluorescent Nanodiamonds as Versatile Intracellular Temperature Sensors.
Temperature is a widely known phenomenon, which plays an extremely important role in biological systems. Its behavior on the macro-scale has been quite well investigated and understood, thanks to the availability of reliable and precise thermometers such as thermocouples and infrared cameras. However, temperature measurements on the subcellular scale present an ongoing challenge due to the absence of universal nanoscale temperature sensors. Recent work on fluorescent nanodiamonds has revealed their unique ability to measure temperature with high spatial and temporal resolution, of particular importance in the intracellular environment. This review summarizes recent progress in the field and highlights the future directions for intracellular temperature sensing using fluorescent nanodiamonds.


The effect of milrinone on hemodynamic and gas exchange parameters in children.
Milrinone is a drug frequently used for hemodynamic support in children during critical illness. Although the hemodynamic changes induced by milrinone in children may appear similar to those of adults, the physiologic contributors of these changes remain vastly unknown. A systematic review was conducted to identify studies characterising the hemodynamic effects of milrinone in children during critical illness for hemodynamic support for various medical conditions. Studies were assessed for quality and those of satisfactory quality with pre- and post-operative hemodynamics for each patient were included in the final analyses. Those not limited to children and those not limited to patients with critical illness were excluded from the final analyses. A total of six studies with 791 patients were included in the final analyses. Milrinone infusion doses ranged from 0.3 to 0.75 mcg/kg/minute with the mean infusion dose being 0.5 mcg/kg/minute. Patients whom received milrinone infusion had greater cardiac output, greater left ventricle shortening fraction, lower right ventricular systolic pressure, and lower serum lactate levels. Systolic blood pressure mean arterial blood pressure and arterial oxygen concentration did not significantly change with administration of milrinone. These results were irrespective of milrinone infusion dose, infusion duration, and study size. Milrinone was found to have several beneficial hemodynamic effects in children during critical illness when used at usual clinical doses.