Complement alternative pathway genetic variation and Dengue infection in the Thai population.
Dengue disease is a mosquito-borne infection caused by Dengue virus. Infection may be asymptomatic or variably manifest as mild Dengue fever (DF) to the most severe form, Dengue haemorrhagic fever (DHF). Mechanisms that influence disease severity are not understood. Complement, an integral component of the immune system, is activated during Dengue infection and the degree of activation increases with disease severity. Activation of the complement alternative pathway is influenced by polymorphisms within activation (factor B rs12614/rs641153, C3 rs2230199) and regulatory [complement factor H (CFH) rs800292] proteins, collectively termed a complotype. Here, we tested the hypothesis that the complotype influences disease severity during secondary Dengue infection. In addition to the complotype, we also assessed two other disease-associated CFH polymorphisms (rs1061170, rs3753394) and a structural polymorphism within the CFH protein family. We did not detect any significant association between the examined polymorphisms and Dengue infection severity in the Thai population. However, the minor allele frequencies of the factor B and C3 polymorphisms were less than 10%, so our study was not sufficiently powered to detect an association at these loci. We were also unable to detect a direct interaction between CFH and Dengue NS1 using both recombinant NS1 and DV2-infected culture supernatants. We conclude that the complotype does not influence secondary Dengue infection severity in the Thai population.


Current management of migraine in US emergency departments: an analysis of the National Hospital Ambulatory Medical Care Survey.
Published data from 1998 revealed that most patients treated for migraine in an emergency department received opioids. Over the intervening years, a large body of evidence has emerged demonstrating the efficacy and safety of non-opioid alternatives. Expert opinion during these years has cautioned against use of opioids for migraine. Our objectives were to compare current frequency of use of various medications for acute migraine in US emergency departments with use of these same medications in 1998 and to identify factors independently associated with opioid use. We analyzed National Hospital Ambulatory Medical Care Survey data from 2010, the most current dataset available. The National Hospital Ambulatory Medical Care Survey is a public dataset collected and distributed by the Centers for Disease Control and Prevention. It is a multi-stage probability sample from randomly selected emergency departments across the country, designed to be representative of all US emergency department visits. We included in our analysis all patients with the ICD9 emergency department discharge diagnosis of migraine. We tabulated frequency of use of specific medications in 2010 and compared these results with the 1998 data. Using a logistic regression model, into which all of the following variables were entered, we explored the independent association between any opioid use in 2010 and sex, age, race/ethnicity, geographic region, type of hospital, triage pain score and history of emergency department use within the previous 12 months. In 2010, there were 1.2 (95% confidence interval 0.9, 1.4) million migraine visits to US emergency departments. Including opioid-containing oral analgesic combinations, opioids were administered in 59% of visits (95% confidence interval 51, 67). The most commonly used parenteral agent, hydromorphone, was used in 25% (95% confidence interval 19, 33) of visits in 2010 versus less than 1% (95% confidence interval 0, 3) in 1998. Conversely, use of meperidine had decreased markedly over the same timeframe. In 2010, it was used in just 7% (95% confidence interval 4, 12) of visits compared to 37% (95% confidence interval 29, 45) in 1998. Metoclopramide, the most commonly used anti-dopaminergic, was administered in 17% (95% confidence interval 12, 23) of visits in 2010 and 3% (95% confidence interval 1, 6) of visits in 1998. Use of any triptan was relatively uncommon in 2010 (7% (95% confidence interval 4, 11) of visits) and in 1998 (10% (95% confidence interval 6, 15) of visits). Of the predictor variables listed above, only emergency department use within the previous 12 months was associated with opioid administration (adjusted odds ratio: 2.87 (95% confidence interval 1.03, 7.97)). In spite of recommendations to the contrary, opioids are still used in more than half of all emergency department visits for migraine. Though use of meperidine has decreased markedly between 1998 and 2010, it has largely been replaced by hydromorphone. Opioid use in migraine visits is independently associated with prior visits to the same emergency department in the previous 12 months.


Prediction of Success in External Cephalic Version under Tocolysis: Still a Challenge.
External cephalic version is a procedure of fetal rotation to a cephalic presentation through manoeuvres applied to the maternal abdomen. There are several prognostic factors described in literature for external cephalic version success and prediction scores have been proposed, but their true implication in clinical practice is controversial. We aim to identify possible factors that could contribute to the success of an external cephalic version attempt in our population. We retrospectively examined 207 consecutive external cephalic version attempts under tocolysis conducted between January 1997 and July 2012. We consulted the department's database for the following variables: race, age, parity, maternal body mass index, gestational age, estimated fetal weight, breech category, placental location and amniotic fluid index. We performed descriptive and analytical statistics for each variable and binary logistic regression. External cephalic version was successful in 46.9% of cases (97/207). None of the included variables was associated with the outcome of external cephalic version attempts after adjustment for confounding factors. We present a success rate similar to what has been previously described in literature. However, in contrast to previous authors, we could not associate any of the analysed variables with success of the external cephalic version attempt. We believe this discrepancy is partly related to the type of statistical analysis performed. Even though there are numerous prognostic factors identified for the success in external cephalic version, care must be taken when counselling and selecting patients for this procedure. The data obtained suggests that external cephalic version should continue being offered to all eligible patients regardless of prognostic factors for success.


Bone-grafting for severe patellar bone loss during revision knee arthroplasty.
Severe patellar bone loss may preclude adequate fixation of another patellar prosthesis as a part of revision knee replacement. The purpose of this study was to describe the surgical technique and early clinical results of an alternative to the conventional treatment options of either patellectomy or retention of the remaining patellar osseous shell. The goals of this procedure are to restore patellar bone stock and potentially to improve the functional outcome. Severe patellar bone loss had left a "patellar shell" that precluded insertion of another patellar implant in nine of 100 consecutive knees undergoing revision total knee arthroplasty. Rather than performing a patellectomy or simply retaining the patellar osseous shell in these nine knees (eight patients), I performed a surgical procedure in which a tissue flap was secured to the patellar rim to contain cancellous bone graft inserted into the patellar bone defect. Final follow-up was at a mean of 36.7 months (range, twenty-four to fifty-five months) after the patellar bone-grafting procedure. The mean preoperative Knee Society scores for function and pain were 39 points (range, 18 to 82 points) and 40 points (range, 20 to 80 points), respectively. At the time of final follow-up, the Knee Society function and pain scores had improved significantly, to a mean function score of 91 points (range, 80 to 98 points) and a mean pain score of 84 points (range, 65 to 100 points) (p<0.05). The point of greatest patellar thickness measured intraoperatively ranged from 7 to 9 mm. Patellar thickness on immediate postoperative Merchant radiographs averaged 22 mm (range, 20 to 25 mm) whereas, at the time of final follow-up, patellar thickness averaged 19.7 mm (range, 17 to 22.5 mm). In contrast with other treatment alternatives, this surgical procedure imparts the potential for restoring patellar bone stock and may improve functional outcome by facilitating patellar tracking and improving quadriceps leverage. On the basis of satisfactory short-term to mid-term clinical results, this technique of patellar bone-grafting appears to be an important addition to the armamentarium of surgeons performing revision knee arthroplasties.


Direct quantitation of growth hormone binding protein in human serum by a ligand immunofunctional assay: comparison with immunoprecipitation and chromatographic methods.
Current methods for measuring GH-binding protein (GHBP) are laborious, require separation of GHBP complex, and may be affected by endogenous GH content of the sample. Such methods estimate binding activity and GHBP concentration can only be obtained by Scatchard analysis. We have developed and validated a 2-site immunofunctional assay for the direct quantitation of GHBP in human serum employing a capture monoclonal antibody against GHBP and a polyclonal antibody against hGH. Results were compared with binding activity data and Scatchard-derived capacity estimates obtained by immunoprecipitation and gel chromatography procedures. Serum samples were obtained from 21 subjects with GH deficiency, 24 patients with acromegaly, and 56 normal subjects; 12 of whom were postmenopausal women who were studied before and during oral estrogen treatment. Using the immunofunctional assay, serum GHBP concentrations in normal subjects ranged from 0.14-3.28 nmol/L, was positively related to body mass index (BMI, P = 0.0004) and negatively related to age (P = 0.015). Women had significantly higher levels (0.99 +/- 0.12 vs. 0.63 +/- 0.09 nmol/L; P = 0.0191) than age and BMI-matched men. GHBP levels were not different between normal, acromegalic, or GH-deficient subjects. Oral estrogen therapy in postmenopausal women increased serum GHBP concentrations up to 5-fold. There was a significant nonlinear relationship between the immunofunctional assay measurements and binding activity by either immunoprecipitation (r = 0.84) or chromatographic (r = 0.73) methods; increase in GHBP concentrations was not reflected in proportionate increase in both activity assays. Estrogen-induced changes in circulating GHBP levels were greatest with the immunofunctional assay followed by Scatchard-derived values from immunoprecipitation and chromatographic methods. We conclude that ligand immunofunctional assay measurements of GHBP are higher but show good agreement with binding activity or Scatchard derived estimates from immunoprecipitation and chromatographic methods. This assay provides a direct and practical tool for rapid, accurate and sensitive estimation of GHBP concentration in serum.


Microbiome interplay: plants alter microbial abundance and diversity within the built environment.
The built indoor microbiome has importance for human health. Residents leave their microbial fingerprint but nothing is known about the transfer from plants. Our hypothesis that indoor plants contribute substantially to the microbial abundance and diversity in the built environment was experimentally confirmed as proof of principle by analyzing the microbiome of the spider plant Chlorophytum comosum in relation to their surroundings. The abundance of Archaea, Bacteria, and Eukaryota (fungi) increased on surrounding floor and wall surfaces within 6 months of plant isolation in a cleaned indoor environment, whereas the microbial abundance on plant leaves and indoor air remained stable. We observed a microbiome shift: the bacterial diversity on surfaces increased significantly but fungal diversity decreased. The majority of cells were intact at the time of samplings and thus most probably alive including diverse Archaea as yet unknown phyllosphere inhabitants. LEfSe and network analysis showed that most microbes were dispersed from plant leaves to the surrounding surfaces. This led to an increase of specific taxa including spore-forming fungi with potential allergic potential but also beneficial plant-associated bacteria, e.g., Paenibacillus. This study demonstrates for the first time that plants can alter the microbiome of a built environment, which supports the significance of plants and provides insights into the complex interplay of plants, microbiomes and human beings.


Pattern ERG correlates of abnormal fundus autofluorescence in patients with retinitis pigmentosa and normal visual acuity.
To examine the functional significance of central abnormalities present in fundus autofluorescence (AF) images in patients with rod-cone dystrophy and good visual acuity. Thirty patients were selected according to three criteria: a clinical diagnosis of retinitis pigmentosa (RP) confirmed with International Society for Clinical Electrophysiology of Vision (ISCEV) standard ERGs, a parafoveal ring of increased high density on fundus AF imaging, and a visual acuity of 20/30 or better. Macular function was assessed with pattern electroretinography (PERG) to checkerboard stimuli of different field sizes. Fundus AF imaging was performed with a confocal scanning laser ophthalmoscope. The radius of the parafoveal ring of high density varied between 1.5 degrees and 9 degrees. The PERG P50 amplitude correlated highly with the radius of the ring of increased autofluorescence (r = 0.80, P < 0.0005, n = 30). PERGs to smaller circular field sizes were present, but increasing field size to beyond that of the high-density autofluorescence ring did not produce further increases in P50 amplitude. There was a high correlation between the minimum stimulus size required to elicit a maximum-amplitude PERG and the radius of the ring (r = 0.87). The high correlation between AF imaging and PERG, an established technique in the assessment of central retinal function, demonstrates the likelihood that autofluorescence abnormalities have functional significance and may therefore be a valuable additional parameter in the monitoring of these patients.


Single versus combination antibiotic therapy in adults hospitalised with community acquired pneumonia.
The benefits of β-lactam/macrolide combination therapy over β-lactam therapy alone for the treatment of hospitalised community-acquired pneumonia (CAP) in relation to pneumonia severity are uncertain. We studied 5240 adults hospitalised with CAP from 72 secondary care trusts across England and Wales. The overall 30-day inpatient (IP) death rate was 24.4%. Combination therapy was prescribed in 3239 (61.8%) patients. In a multivariable model, combination therapy was significantly associated with lower 30-day IP death rate in patients with moderate-severity CAP (adjusted OR 0.54, 95% CI 0.41 to 0.72) and high-severity CAP (adjusted OR 0.76, 95% CI 0.60 to 0.96) but not low-severity CAP.


Discordant Genome Assemblies Drastically Alter the Interpretation of Single-Cell RNA Sequencing Data Which Can Be Mitigated by a Novel Integration Method.
Advances in sequencing and assembly technology have led to the creation of genome assemblies for a wide variety of non-model organisms. The rapid production and proliferation of updated, novel assembly versions can create vexing problems for researchers when multiple-genome assembly versions are available at once, requiring researchers to work with more than one reference genome. Multiple-genome assemblies are especially problematic for researchers studying the genetic makeup of individual cells, as single-cell RNA sequencing (scRNAseq) requires sequenced reads to be mapped and aligned to a single reference genome. Using the Astyanax mexicanus, this study highlights how the interpretation of a single-cell dataset from the same sample changes when aligned to its two different available genome assemblies. We found that the number of cells and expressed genes detected were drastically different when aligning to the different assemblies. When the genome assemblies were used in isolation with their respective annotations, cell-type identification was confounded, as some classic cell-type markers were assembly-specific, whilst other genes showed differential patterns of expression between the two assemblies. To overcome the problems posed by multiple-genome assemblies, we propose that researchers align to each available assembly and then integrate the resultant datasets to produce a final dataset in which all genome alignments can be used simultaneously. We found that this approach increased the accuracy of cell-type identification and maximised the amount of data that could be extracted from our single-cell sample by capturing all possible cells and transcripts. As scRNAseq becomes more widely available, it is imperative that the single-cell community is aware of how genome assembly alignment can alter single-cell data and their interpretation, especially when reviewing studies on non-model organisms.


Examination of acetabular labral tear: a continued diagnostic challenge.
Acetabular labrum tears (ALT) are present in 22-55% of individuals with hip or groin pain. Tears can occur as a result of trauma or degeneration and are markedly associated with femoral acetabular morphological variations. An ALT can lead to biomechanical deficiencies and a loss of stability to the coxafemoral joint due to the labrum serving as a stabilising structure of this joint. The diagnosis of ALT is complex and multidimensional. Although tremendous improvements in diagnostic utility for ALT have occurred in the past 25 years, there are few patient history, clinical examination and special test findings that are unique to the condition. Imaging methods such as MRI, CT and ultrasonography have demonstrated reasonable accuracy, but not at a level that allows use as a stand-alone measure. Outcomes measures that focus on functional limitation or that are used to measure recovery should envelop the complexities of the condition and be captured using both self-report and physical performance measures. Only when patient history, objective testing, clinical examination special testing and imaging are combined can a clinician fully elucidate the multidimensional diagnosis of ALT.


Recommendations for improved data processing from expired gas analysis indirect calorimetry.
There is currently no universally recommended and accepted method of data processing within the science of indirect calorimetry for either mixing chamber or breath-by-breath systems of expired gas analysis. Exercise physiologists were first surveyed to determine methods used to process oxygen consumption ((.)VO2) data, and current attitudes to data processing within the science of indirect calorimetry. Breath-by-breath datasets obtained from indirect calorimetry during incremental exercise were then used to demonstrate the consequences of commonly used time, breath and digital filter post-acquisition data processing strategies. Assessment of the variability in breath-by-breath data was determined using multiple regression based on the independent variables ventilation (VE), and the expired gas fractions for oxygen and carbon dioxide, FEO2 and FECO2, respectively. Based on the results of explanation of variance of the breath-by-breath (.)VO2 data, methods of processing to remove variability were proposed for time-averaged, breath-averaged and digital filter applications. Among exercise physiologists, the strategy used to remove the variability in (.)VO2 measurements varied widely, and consisted of time averages (30 sec [38%], 60 sec [18%], 20 sec [11%], 15 sec [8%]), a moving average of five to 11 breaths (10%), and the middle five of seven breaths (7%). Most respondents indicated that they used multiple criteria to establish maximum ((.)VO2 ((.)VO2max) including: the attainment of age-predicted maximum heart rate (HR(max)) [53%], respiratory exchange ratio (RER) >1.10 (49%) or RER >1.15 (27%) and a rating of perceived exertion (RPE) of >17, 18 or 19 (20%). The reasons stated for these strategies included their own beliefs (32%), what they were taught (26%), what they read in research articles (22%), tradition (13%) and the influence of their colleagues (7%). The combination of VE, FEO2 and FECO2 removed 96-98% of (.)VO2 breath-by-breath variability in incremental and steady-state exercise (.)VO2 data sets, respectively. Correction of residual error in (.)VO2 datasets to 10% of the raw variability results from application of a 30-second time average, 15-breath running average, or a 0.04 Hz low cut-off digital filter. Thus, we recommend that once these data processing strategies are used, the peak or maximal value becomes the highest processed datapoint. Exercise physiologists need to agree on, and continually refine through empirical research, a consistent process for analysing data from indirect calorimetry.


One-year follow-up of platelet-rich plasma treatment in chronic Achilles tendinopathy: a double-blind randomized placebo-controlled trial.
Achilles tendinopathy is a common disease among both athletes and in the general population in which the use of platelet-rich plasma has recently been increasing. Good evidence for the use of this autologous product in tendinopathy is limited, and data on longer-term results are lacking. To study the effects of a platelet-rich plasma injection in patients with chronic midportion Achilles tendinopathy at 1-year follow-up. Randomized controlled trial; Level of evidence, 1. Fifty-four patients, aged 18 to 70 years, with chronic tendinopathy 2 to 7 cm proximal to the Achilles tendon insertion were randomized to receive either a blinded injection containing platelet-rich plasma or saline (placebo group) in addition to an eccentric training program. The main outcome was the validated Victorian Institute of Sports Assessment-Achilles score. Patient satisfaction was recorded and ultrasound examination performed at baseline and follow-up. The mean Victorian Institute of Sports Assessment-Achilles score improved in both the platelet-rich plasma group and the placebo group after 1 year. There was no significant difference in increase between both groups (adjusted between-group difference, 5.5; 95% confidence interval, -4.9 to 15.8, P = .292). In both groups, 59% of the patients were satisfied with the received treatment. Ultrasonographic tendon structure improved significantly in both groups but was not significantly different between groups (adjusted between-group difference, 1.2%; 95% confidence interval, -4.1 to 6.6, P = .647). This randomized controlled trial showed no clinical and ultrasonographic superiority of platelet-rich plasma injection over a placebo injection in chronic Achilles tendinopathy at 1 year combined with an eccentric training program.


Multigenic system controlling viral systemic infection determined by the interactions between Cucumber mosaic virus genes and quantitative trait loci of soybean cultivars.
Soybean 'Harosoy' is resistant to Cucumber mosaic virus soybean strain C (CMV-SC) and susceptible to CMV-S strain D (CMV-SD). Using enzyme-linked immunosorbent assay and Northern hybridization, we characterized the Harosoy resistance and found that CMV-SC did not spread systemically but was restricted to the inoculated leaves in Harosoy. Harosoy resistance was not controlled by either a dominant or recessive single gene. To dissect this system controlling long-distance movement of CMV in soybean, we constructed infectious cDNA clones of CMV-SC and CMV-SD. Using these constructs and the chimeric RNAs, we demonstrated that two viral components were required for systemic infection by the virus. The region including the entire 2b gene and the 5' region of RNA3 (mainly the 5' untranslated region) together were required. By quantitative trait locus (QTL) analysis using an F(2) population and the F(3) families derived from Harosoy and susceptible 'Nemashirazu', we also showed that at least three QTLs affected systemic infection of CMV in soybean. Our study on Harosoy resistance to CMV-SC revealed an interesting mechanism, in which multiple host and viral genes coordinately controlled viral systemic infection.


Item development and validity testing for a self- and proxy report: the safe driving behavior measure.
We report on item development and validity testing of a self-report older adult safe driving behaviors measure (SDBM). On the basis of theoretical frameworks (Precede-Proceed Model of Health Promotion, Haddon's matrix, and Michon's model), existing driving measures, and previous research and guided by measurement theory, we developed items capturing safe driving behavior. Item development was further informed by focus groups. We established face validity using peer reviewers and content validity using expert raters. Peer review indicated acceptable face validity. Initial expert rater review yielded a scale content validity index (CVI) rating of 0.78, with 44 of 60 items rated > or = 0.75. Sixteen unacceptable items (< or = 0.5) required major revision or deletion. The next CVI scale average was 0.84, indicating acceptable content validity. The SDBM has relevance as a self-report to rate older drivers. Future pilot testing of the SDBM comparing results with on-road testing will define criterion validity.


Evolution and phylogeny of the mud shrimps (Crustacea: Decapoda) revealed from complete mitochondrial genomes.
The evolutionary history and relationships of the mud shrimps (Crustacea: Decapoda: Gebiidea and Axiidea) are contentious, with previous attempts revealing mixed results. The mud shrimps were once classified in the infraorder Thalassinidea. Recent molecular phylogenetic analyses, however, suggest separation of the group into two individual infraorders, Gebiidea and Axiidea. Mitochondrial (mt) genome sequence and structure can be especially powerful in resolving higher systematic relationships that may offer new insights into the phylogeny of the mud shrimps and the other decapod infraorders, and test the hypothesis of dividing the mud shrimps into two infraorders. We present the complete mitochondrial genome sequences of five mud shrimps, Austinogebia edulis, Upogebia major, Thalassina kelanang (Gebiidea), Nihonotrypaea thermophilus and Neaxius glyptocercus (Axiidea). All five genomes encode a standard set of 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes and a putative control region. Except for T. kelanang, mud shrimp mitochondrial genomes exhibited rearrangements and novel patterns compared to the pancrustacean ground pattern. Each of the two Gebiidea species (A. edulis and U. major) and two Axiidea species (N. glyptocercus and N. thermophiles) share unique gene order specific to their infraorders and analyses further suggest these two derived gene orders have evolved independently. Phylogenetic analyses based on the concatenated nucleotide and amino acid sequences of 13 protein-coding genes indicate the possible polyphyly of mud shrimps, supporting the division of the group into two infraorders. However, the infraordinal relationships among the Gebiidea and Axiidea, and other reptants are poorly resolved. The inclusion of mt genome from more taxa, in particular the reptant infraorders Polychelida and Glypheidea is required in further analysis. Phylogenetic analyses on the mt genome sequences and the distinct gene orders provide further evidences for the divergence between the two mud shrimp infraorders, Gebiidea and Axiidea, corroborating previous molecular phylogeny and justifying their infraordinal status. Mitochondrial genome sequences appear to be promising markers for resolving phylogenetic issues concerning decapod crustaceans that warrant further investigations and our present study has also provided further information concerning the mt genome evolution of the Decapoda.


WHO BENEFITS MOST FROM A BROADLY TARGETED PREVENTION PROGRAM? DIFFERENTIAL EFFICACY ACROSS POPULATIONS IN THE TEEN OUTREACH PROGRAM.
This study examined a highly successful, well-documented, national program to prevent teenage pregnancy and school failure-the Teen Outreach program-to address a fundamental question: How well can a developmentally focused, broadly targeted prevention program address the needs of those students within the program who are at the highest risk of problematic behavior. The hypothesis that the developmental focus of a broadly targeted intervention would lead it to have greater program efficacy among those young people who began the program at greatest risk was examined with multisite data collected on more than 3,300 Teen Outreach and comparison group students. Results confirmed prior findings regarding the overall efficacy of the Teen Outreach program, and indicated that the program appeared most effective for those students at greatest initial risk of the problem behaviors being targeted. Implications for the targeting of the Teen Outreach program specifically and of similar primary prevention programs more generally are discussed.


Ixodes scapularis and Borrelia burgdorferi among diverse habitats within a natural area in east-central Illinois.
The distributions of the tick vector, Ixodes scapularis, and of the etiologic agent of Lyme disease, Borrelia burgdorferi (Bb), have continued expanding in Illinois over the past 20 years, but the extent of their spread is not well known. The role of multiple habitats in the establishment and maintenance of I. scapularis and Bb at local scales is not well understood, and the use of integrated approaches to evaluate local scale dynamics is rare. We evaluated habitat diversity and temporal changes of I. scapularis occurrence and Bb infection within a natural area in Piatt County, Illinois, where I. scapularis were first detected in 2002. Small mammals were trapped and attached ticks were collected in young forest, prairie, mature forest, and flood plain sites from 2005 to 2009. Small mammal abundance, and the prevalence (% mammals infested), mean intensity (I. scapularis per infested mammal), and relative density (I. scapularis per mammal trapped) of I. scapularis were computed for each habitat type and compared. Immature I. scapularis were tested for Bb infection using polymerase chain reaction techniques. Out of 2446 trapped small mammals, 388 were infested with I. scapularis. The prairie had the highest diversity of small mammal hosts. Prevalence, mean intensity, and relative density of I. scapularis and prevalence of Bb infection were highest for the prairie and young forest sites; in the former, all infection was associated with the prairie vole, Microtus ochrogaster. The minimum Bb infection prevalence of on-host I. scapularis collected in the natural area was 14% (n=56). Unlike previous studies solely focused on forested areas and Peromyscus leucopus, our study is the first to provide evidence of I. scapularis collected from prairie habitat and other reservoir hosts, particularly M. ochrogaster.


Postoperative increase in grey matter volume in visual cortex after unilateral cataract surgery.
The developing visual cortex has a strong potential to undergo plastic changes. Little is known about the potential of the ageing visual cortex to express plasticity. A pertinent question is whether therapeutic interventions can trigger plastic changes in the ageing visual cortex by restoring vision. Twelve patients aged 50-85 years underwent structural high-resolution T1-weighted MRI of the whole brain 2 days and 6 weeks after unilateral cataract surgery. Voxel-based morphometry (VBM) based on T1-weighted magnetic resonance imaging (MRI) was employed to test whether cataract surgery induces a regional increase in grey matter in areas V1 and V2 of the visual cortex. In all patients, cataract surgery immediately improved visual acuity, contrast sensitivity and mean sensitivity in the visual field of the operated eye. The improvement in vision was stable throughout the 6 weeks after operation. VBM revealed a regional expansion of grey matter volume in area V2 contralateral to the operated eye during the 6-week period after surgery. Individual increases in grey matter were predicted by the symmetry in visual acuity between the operated eye and nonoperated eye. The more symmetrical visual acuity became after unilateral cataract surgery, the more pronounced was the grey matter increase in visual cortex. The data suggest that cataract surgery triggered a use-dependent structural plasticity in V2 presumably through improved binocular integration of visual input from both eyes. We conclude that activity-dependent cortical plasticity is preserved in the ageing visual cortex and may be triggered by restoring impaired vision.


Physiological and biochemical alterations induced in the mussel Mytilus galloprovincialis after short and long-term exposure to carbamazepine.
The bivalve Mytilus galloprovincialis collected in the Ria de Aveiro, was selected to evaluate the acute and chronic effects of carbamazepine (CBZ) at environmentally relevant concentrations. CBZ is an antiepileptic drug widely found in the aquatic environment with toxic effects to inhabiting organisms. However, few studies evaluated the acute and chronic toxicity of this drug. The experiment was performed by exposing mussels to 0.0, 0.3, 3.0, 6.0 and 9.0 CBZ μg/L, for 96 h and 28 days. To assess the toxicity of the drug, a battery of biomarkers related to mussels general physiological health status and oxidative stress was applied. CBZ was quantified in mussel tissues by an Enzyme-Linked Immunosorbent Assay (ELISA). The results obtained show that CBZ did not induce oxidative stress. However, our findings demonstrated that the drug was taken up by mussels even though presenting low bioconcentration factor (BCF) values (up to 2.2). Furthermore, our results demonstrated that after a chronic exposure the physiological parameters, namely the condition and gonadosomatic indices, were negatively affected which may impair organisms' reproductive capacity with consequences to population sustainability.


Inflammation During Gestation Induced Spatial Memory and Learning Deficits: Attenuated by Physical Exercise in Juvenile Rats.
Gestational infections induced inflammation (GIII) is a cause of various postnatal neurological deficits in developing countries. Such intra uterine insults could result in persistent learning-memory disabilities. There are no studies elucidating the efficacy of adolescence exercise on spatial learning- memory abilities of young adult rats pre-exposed to inflammatory insult during fetal life. The present study addresses the efficacy of physical (running) exercise during adolescent period in attenuating spatial memory deficits induced by exposure to GIII in rats. Pregnant Wistar dams were randomly divided into control and lipopolysaccharide (LPS) groups, injected intra peritoneally (i.p) with saline (0.5ml) or lipopolysaccharide (LPS) (0.5mg/kg) on alternate days from gestation day 14 (GD 14) till delivery. After parturition, pups were divided into 3 groups (n=6/group) a) Sham control and LPS group divided into 2 subgroups- b) LPS and c) LPS exercise group. Running exercise was given only to LPS exercise group during postnatal days (PNDs) 30 to 60 (15min/day). Spatial learning and memory performance was assessed by Morris water maze test (MWM), on postnatal day 61 to 67 in all groups. Young rats pre-exposed to GIII and subjected to running exercise through juvenile period displayed significant decrease in latency to reach escape platform and spent significant duration in target quadrant in MWM test, compared to age matched LPS group. Results of the current study demonstrated that exercise through juvenile/adolescent period effectively mitigates gestational inflammation-induced cognitive deficits in young adult rats. Inflammation during gestation impairs offspring's spatial memory and learning abilities. Whereas, early postnatal physical exercise attenuates, to higher extent, cognitive impairment resulted from exposure to LPS induced inflammation during intrauterine growth period.


Development of the Successful Transitions Assessment Tool (STAT) for Crossover Youth (COY): Youth voice and expert feedback with a Delphi approach.
Crossover youth (COY) are involved with child welfare and/or the justice system and experience multifaceted challenges in their transition into adulthood. A literature review identified eight critical indicators of successful transitions for COY and the absence of a validated comprehensive assessment that is youth informed and culturally sensitive. To develop a Successful Transitions Assessment Tool (STAT) that is informed by research, Critical Race Theory, and subject matter experts. Field experts were approached to review the initial STAT, developed from literature findings, using the Delphi approach; 13 experts completed the first round and nine completed the second round. Child welfare youth participated in a focus group and youth justice youth completed interviews; 10 youth aged 16 to 24 provided feedback. A two-round Delphi study with experts used a survey link. The focus groups and interviews with youth experts were conducted virtually. Expert rankings in the examined areas were aggregated and qualitative data were thematically analyzed. Responding experts in both rounds of the Delphi process ranked the STAT highly, achieving scores of 16.8 and 16.4 of 20 possible points, respectively (consensus >80 %). No additional rounds were deemed necessary. All suggested tool clarifications were incorporated. Additional questions were added based on suggestions relevant to the eight critical domains. This innovative study created a single, brief, yet comprehensive assessment tool of eight key domains for successful transitions out of care settings, informed by experts through the Delphi approach and youth themselves.


Triglyceride Glucose Index as a Surrogate Measure of Insulin Sensitivity in a Caucasian Pediatric Population
The triglyceride and glucose (TyG) index has been proposed as a simple surrogate of insulin resistance (IR) with high sensitivity as an IR index besides the well known homeostasis model assessment of IR (HOMA-IR). Limited data are reported in children. We investigated the sensitivity and specificity of TyG index in a pediatric Caucasian population, as a surrogate measure of IR and compared the results with HOMA-IR. We enrolled 541 children (11.7±2.71 yrs). According to body mass index (BMI) chart, the subjects were divided into three groups: normal weight BMI<75th percentile, overweight BMI 75th–95th percentile, and obese>95th percentile. TyG index was calculated as (ln[fasting triglycerides(mg/dl)×fasting plasma glucose(mg/dl)/2]) and considered pathological when exceeding 7.88. HOMA-IR was calculated as (insulin×glucose)/22.5 and defined pathological whenever exceeding 97.5th percentile for age and sex. In children with overweight/obesity TyG index was higher compared to normal weight subjects (p<0.001). TyG index was correlated with BMI (p<0.001); WHtR (p<0.001), total and HDL cholesterol (p<0.001); ALT (p<0.001), blood pressure (p<0.001). A correlation between TyG index and HOMAIR (p<0.001) as well as high TyG index and pathological HOMA-IR (p<0.001) were noted. The optimal cut-off for IR was considered 7.98 (sensitivity 60%; specificity 78%; AUC 0.69). TyG index is a useful and cost-effective index of IR among children and adolescents. The cutoff 7.98 may be used for IR risk screening in childhood obesity, but we recommend caution when used in other populations.


Intracellular accumulation and immunological responses of lipid modified magnetic iron nanoparticles in mouse antigen processing cells.
Understanding the effects of magnetic iron nanoparticles (MINPs) on the immune response is vitally important for biomedical applications such as cancer therapy, disease diagnosis and novel cancer imaging. In this study, lipid modified MINPs were designed and prepared by introducing the neutral lipid DSPE-PEG or the zwitterionic lipid DSPE-PCB into hydrophobic MINPs through hydrophobic interaction (L-MINPs and ZL-MINPs, respectively). The effect of L-MINPs and ZL-MINPs on the intracellular accumulation and immune responses of three kinds of antigen processing cells was examined. The results indicated that the high cellular uptake efficiency of surface coated MINPs was strongly related to the nature of the coating lipid, with the zwitterionic lipid being more effective than PEGylated ones. Besides, the results from flow cytometry (FCM), confocal laser scanning microscopy (CLSM) and Prussian blue staining demonstrated a time- and concentration-dependent MINP internalization. The uptake of zwitterionic lipid modified MINPs (ZL-MINPs) induced very low cytotoxicity and a strong mixed Th1/Th2 type immune response. L-MINPs could induce a strong increase in pro-inflammatory cytokines with a slight secretion of Th2 cytokines. Besides, no IL-10 was observed in both groups, indicating that MINPs with lipid modification were absence of immunosuppression. In conclusion, this study addresses an important implication of the lipid type and Fe concentration on the immune stimulation of cells and supports the potential for further development of biomedical applications.


Efficacy and safety of radotinib in chronic phase chronic myeloid leukemia patients with resistance or intolerance to BCR-ABL1 tyrosine kinase inhibitors.
Radotinib (IY5511HCL), a novel and selective BCR-ABL1 tyrosine kinase inhibitor, has shown pre-clinical and phase I activity and safety in chronic myeloid leukemia. This phase II study investigated the efficacy and safety of radotinib in Philadelphia chromosome-positive chronic phase-chronic myeloid leukemia patients with resistance and/or intolerance to BCR-ABL1 tyrosine kinase inhibitors. Patients received radotinib 400 mg twice daily for 12 cycles based on results from the phase I trial. The primary end point was rate of major cytogenetic response by 12 months. A total of 77 patients were enrolled. Major cytogenetic response was achieved in 50 (65%; cumulative 75%) patients, including 36 (47%) patients with complete cytogenetic response by 12 months. Median time to major cytogenetic response and complete cytogenetic response were 85 days and 256 days, respectively. Major cytogenetic response and complete cytogenetic response rates were similar between imatinib-resistant and imatinib-intolerant patients, but were higher in patients without BCR-ABL1 mutations. Overall and progression-free survival rates at 12 months were 96.1% and 86.3%, respectively. All newly-occurring or worsening grade 3/4 hematologic abnormalities included thrombocytopenia (24.7%) and anemia (5.2%); grade 3/4 drug-related non-hematologic adverse events included fatigue (3.9%), asthenia (3.9%), and nausea (2.6%). The most common biochemistry abnormality was hyperbilirubinemia (grade 3/4 23.4%), and 12 of 18 cases were managed with dose modification. Study findings suggest radotinib is effective and well tolerated in chronic phase-chronic myeloid leukemia patients with resistance and/or intolerance to BCR-ABL1 tyrosine kinase inhibitors and may represent a promising alternative for these patients. (clinicaltrials.gov identifier: 01602952).


Adrenaline Stimulates Glucagon Secretion by Tpc2-Dependent Ca2+ Mobilization From Acidic Stores in Pancreatic α-Cells.
Adrenaline is a powerful stimulus of glucagon secretion. It acts by activation of β-adrenergic receptors, but the downstream mechanisms have only been partially elucidated. Here, we have examined the effects of adrenaline in mouse and human α-cells by a combination of electrophysiology, imaging of Ca2+ and PKA activity, and hormone release measurements. We found that stimulation of glucagon secretion correlated with a PKA- and EPAC2-dependent (inhibited by PKI and ESI-05, respectively) elevation of [Ca2+]i in α-cells, which occurred without stimulation of electrical activity and persisted in the absence of extracellular Ca2+ but was sensitive to ryanodine, bafilomycin, and thapsigargin. Adrenaline also increased [Ca2+]i in α-cells in human islets. Genetic or pharmacological inhibition of the Tpc2 channel (that mediates Ca2+ release from acidic intracellular stores) abolished the stimulatory effect of adrenaline on glucagon secretion and reduced the elevation of [Ca2+]i Furthermore, in Tpc2-deficient islets, ryanodine exerted no additive inhibitory effect. These data suggest that β-adrenergic stimulation of glucagon secretion is controlled by a hierarchy of [Ca2+]i signaling in the α-cell that is initiated by cAMP-induced Tpc2-dependent Ca2+ release from the acidic stores and further amplified by Ca2+-induced Ca2+ release from the sarco/endoplasmic reticulum.


Non-attendance at psychiatric outpatient clinics: communication and implications for primary care.
What should happen when an outpatient fails to attend a psychiatric clinic? Guidelines from the General Medical Services Committee suggest that general practitioners (GPs) have no further responsibility of care for a patient once a referral to a psychiatrist has been made. This raises questions about the formulation of effective management plans for those patients with whom psychiatric services find it difficult to engage due to non-compliance with assessment and follow-up. To investigate communication between GPs, patients, and psychiatrists at referral and following attendance or non-attendance at outpatient appointments. A prospective study of a random sample of attenders and non-attenders at psychiatric outpatient clinics. Patients and GPs were interviewed to obtain data about the referral process. GPs' views on communication from psychiatrists and the appropriate course of action following clinic non-attendance were investigated. The quality of referral and clinic letters for attenders and non-attenders was compared. There was no difference in the quality of referral letter for attenders and non-attenders. Psychiatrists were less likely to write to GPs about follow-up patients' appointments than new patients' appointments; communication was least likely when a follow-up patient missed their appointment. GPs considered follow-up non-attenders were more likely to need a further appointment than new patient non-attenders, but did not identify a role for themselves in engaging with follow-up non-attenders. Communication between GPs and psychiatrists about new patients seems adequate. However, there are important deficits in communication from psychiatrists to GPs about follow-up patients, especially non-attenders who are often more severely ill and difficult to engage. An effective response for this group is likely to need cooperative health and social service action rather than rigid guidelines concerning clinical responsibility.


In vitro anticancer activity and biologically relevant metabolization of organometallic ruthenium complexes with carbohydrate-based ligands.
The synthesis and in vitro anticancer activity of dihalogenido(eta6-p-cymene)(3,5,6-bicyclophosphite-alpha-D-glucofuranoside)ruthenium(II) complexes are described. The compounds were characterized by NMR spectroscopy and ESI mass spectrometry, and the molecular structures of dichlorido-, dibromido- and diiodido(eta6-p-cymene)(3,5,6-bicyclophosphite-1,2-O-isopropylidene-alpha-D-glucofuranoside)ruthenium(II) were determined by X-ray diffraction analysis. The complexes were shown to undergo aquation of the first halido ligand in aqueous solution, followed by hydrolysis of a P--O bond of the phosphite ligand, and finally formation of dinuclear species. The hydrolysis mechanism was confirmed by DFT calculations. The aquation of the complexes was markedly suppressed in 100 mM NaCl solution, and notably only very slow hydrolysis of the P--O bond was observed. The complexes showed affinity towards albumin and transferrin and monoadduct formation with 9-ethylguanine. In vitro studies revealed that the 3,5,6-bicyclophosphite-1,2-O-cyclohexylidene-alpha-D-glucofuranoside complex is the most cytotoxic compound in human cancer cell lines (IC50 values from 30 to 300 microM depending on the cell line).


Erosion of microbicide formulation coating layers: effects of contact and shearing with vaginal fluid or semen.
An effective vaginal microbicide formulation must distribute and maintain an epithelial coating layer. The post-application durability of this coating is significantly affected by the vaginal environment. A new in vitro assay quantified coating layer erosion after contact and shear with simulated vaginal fluid or semen. Coating layer persistence and viscosity of both fluid and gel layers were assessed versus time. Five vaginal formulations were studied. In all gels, there was an overall trend of rapid ( approximately 30 min) and significant viscosity loss. Although there were differences across gels and between simulants, greater erosion occurred after contact with the low-pH vaginal fluid simulant (>50% viscosity decrease), as compared to an alkaline semen simulant. These in vitro results suggest significant differences in vivo of vaginal coating retention by the test gels. This new assay can be diversified to create a spectrum of biologically relevant conditions which collectively simulate the natural history of vaginal formulation residence.


The Salmonella effector protein PipB2 is a linker for kinesin-1.
Understanding the mechanisms of Salmonella virulence is an important challenge. The capacity of this intracellular bacterial pathogen to cause diseases depends on the expression of virulence factors including the second type III secretion system (TTSS-2), which is used to translocate into the eukaryotic cytosol a set of effector proteins that divert the biology of the host cell and shape the bacterial replicative niche. Yet little is known about the eukaryotic functions affected by individual Salmonella effectors. Here we report that the TTSS-2 effector PipB2 interacts with the kinesin light chain, a subunit of the kinesin-1 motor complex that drives anterograde transport along microtubules. Translocation of PipB2 is both necessary and sufficient for the recruitment of kinesin-1 to the membrane of the Salmonella-containing vacuole. In vivo, PipB2 contributes to the attenuation of Salmonella mutant strains in mice. Taken together, our data indicate that the TTSS-2-mediated fine-tuning of kinesin-1 activity associated with the bacterial vacuole is crucial for the virulence of Salmonella.


Drop mobility on superhydrophobic microstructured surfaces with wettability contrasts.
Manipulation of drop motion has attracted considerable attention recently as it is pertinent to industrial/biological applications such as microfluidics. Wettability gradients/contrasts applied to microtextured, superhydrophobic surfaces are probable candidates for engineering drop motion by virtue of their wettability controllability and low contact angle hysteresis. In the present work, we present a systematic study of drop mobility induced via wettability contrasts. A millimetre-sized water drop, placed on the boundary between two surfaces with distinct, uniform arrays of pillars, immediately moved toward the surface more densely populated with asperities, which was relatively more hydrophilic. The velocity of the motion was found to increase proportionally with the difference in pillar densities on each surface, in circumstances where the rear side surface had sufficiently small contact angle hysteresis. To elucidate the underlying mechanism of drop motion, we implemented a surface energy analysis for each motion event. Motion was initiated by the excess surface free energy due to drop deformation and directed in favour of energy minimisation. Lastly, we propose a theory to predict the direction of the drop which at the same time acts as the criterion for the motion to ensue.


Revisiting the effect of referral bias on the clinical spectrum of infective endocarditis in adults.
Referral bias occurs because of the clustering of patients at tertiary care centers. This may result in the distortion of observed clinical manifestations of rare diseases. This analysis evaluates the effect of referral bias on the epidemiology of infective endocarditis (IE) in the International Collaboration on Endocarditis-Prospective Cohort Study (ICE-PCS). This is a prospective multicenter cohort study comparing transferred and non-transferred patients with IE. Factors independently associated with transfer status were evaluated using multivariable logistic regression. A total of 2,760 patients were included in the analysis, of which 1,164 (42.2%) were transferred from other medical centers. Transferred patients more often underwent surgery for IE (odds ratio [OR] = 2.5; 95% confidence interval [CI] 1.9-3.2). They were also more likely to have complications such as stroke (OR = 1.5; 95% CI 1.3-1.9), heart failure (OR = 1.4; 95% CI 1.1-1.6), and new valvular regurgitation (OR = 1.3; 95% CI 1.1-1.6). The in-hospital mortality rates were similar in both groups. Patients with IE who require surgery and suffer complications are referred to tertiary hospitals more frequently than patients with an uncomplicated course. Hospital transfer has no obvious effect on the in-hospital mortality. Referral bias should be taken into consideration when describing the clinical spectrum of IE.


Optimal strategy for fabrication of large aperture aspheric surfaces.
Aspheric surfaces are widely used because of their desirable characteristics. Such a surface can obtain nearly perfect imaging quality with fewer optical elements and reduce the size and mass of optical systems. Various machine systems have been developed based on modern deterministic polishing technologies for large aperture aspheric surfaces. Several factors affect the final precision of large aperture aspheric surfaces, such as the velocity limit of the machine and the path design. Excess velocity, which will be truncated automatically by the computer numerical control system, may cause the dwell time to deviate from the desired time. When a path designed on a two-dimensional surface map with equidistant pitch is projected onto an aspheric surface, the pitch changes as a result of the varied curvature of the aspheric surface. This may affect the removal map and cause some ripple errors. A multiregion distribution strategy, which includes velocity checking, is proposed in this study to avoid exceeding the velocity limits. The strategy can be used to modify local errors and edge effects. A three-dimensional spiral path generation method is also presented using an iterative method to ensure uniformity in the space length of the adjacent circle of the spiral path. This process can reduce the ripple error caused by the overlapping of tool paths. A polishing experiment was conducted, and the results proved the validity of the proposed strategies.


3D-footprint: a database for the structural analysis of protein-DNA complexes.
3D-footprint is a living database, updated and curated on a weekly basis, which provides estimates of binding specificity for all protein-DNA complexes available at the Protein Data Bank. The web interface allows the user to: (i) browse DNA-binding proteins by keyword; (ii) find proteins that recognize a similar DNA motif and (iii) BLAST similar DNA-binding proteins, highlighting interface residues in the resulting alignments. Each complex in the database is dissected to draw interface graphs and footprint logos, and two complementary algorithms are employed to characterize binding specificity. Moreover, oligonucleotide sequences extracted from literature abstracts are reported in order to show the range of variant sites bound by each protein and other related proteins. Benchmark experiments, including comparisons with expert-curated databases RegulonDB and TRANSFAC, support the quality of structure-based estimates of specificity. The relevant content of the database is available for download as flat files and it is also possible to use the 3D-footprint pipeline to analyze protein coordinates input by the user. 3D-footprint is available at http://floresta.eead.csic.es/3dfootprint with demo buttons and a comprehensive tutorial that illustrates the main uses of this resource.


Hierarchical cognitive control deficits following damage to the human frontal lobe.
Cognitive control permits us to make decisions about abstract actions, such as whether to e-mail versus call a friend, and to select the concrete motor programs required to produce those actions, based on our goals and knowledge. The frontal lobes are necessary for cognitive control at all levels of abstraction. Recent neuroimaging data have motivated the hypothesis that the frontal lobes are organized hierarchically, such that control is supported in progressively caudal regions as decisions are made at more concrete levels of action. We found that frontal damage impaired action decisions at a level of abstraction that was dependent on lesion location (rostral lesions affected more abstract tasks, whereas caudal lesions affected more concrete tasks), in addition to impairing tasks requiring more, but not less, abstract action control. Moreover, two adjacent regions were distinguished on the basis of the level of control, consistent with previous functional magnetic resonance imaging results. These results provide direct evidence for a rostro-caudal hierarchical organization of the frontal lobes.


Nonregenerative axonal growth within the mature mammalian brain: ultrastructural identification of sympathohippocampal sprouts.
Damage to septohippocampal neurons in the adult rat results in sprouting of sympathetic axons into the denervated hippocampal formation. However, the distribution of sympathohippocampal fibers has only been assessed with light microscopic techniques, and it is not known if the sprouted fibers leave the blood vessels, along which they migrate into the hippocampal formation, to enter the hippocampal neuropil and, if they do, whether they form synaptic contacts with central neurons. Using the tetramethylbenzidine technique to visualize anterogradely transported wheat germ agglutinin-horseradish peroxidase conjugate, we identified sprouted sympathetic fibers in the hippocampal formation at both the light and electron microscopic level in albino rats receiving medial septal lesions. The majority of labeled fibers were observed within the regions immediately above and below the granule cell layer. Although most of the labeled sprouts were observed in association with intraparenchymal blood vessels, where they were usually apposed to the basal lamina, approximately a third of the labeled profiles were present within the neuropil with no obvious vascular relationships. Most of the profiles were identified as unmyelinated axons or vesicle-filled varicosities. Many of the latter structures contained small dense-cored vesicles, but in our sample none of the labeled profiles were observed to form membrane specializations with adjacent structures, and many were partly surrounded by presumed astrocytic processes. These results document the invasion of the CNS by sprouting axons of peripheral origin indicating that axonal elongation from uninjured neurons can occur within the mature mammalian CNS under certain circumstances. In addition, the presence of significant numbers of sympathetic fibers within the hippocampal neuropil indicates that they may be in a strategic position to influence hippocampal function.


Conditional recurrence-free survival of clinical complete responders managed by watch and wait after neoadjuvant chemoradiotherapy for rectal cancer in the International Watch & Wait Database: a retrospective, international, multicentre registry study.
Watch and wait is a novel management strategy in patients with rectal cancer who have a clinical complete response after neoadjuvant chemoradiotherapy. Surveillance of these patients is generally intensive, because local regrowth (with the potential for salvage) occurs in 25% of patients, and distant metastases occur in 10% of patients. It is unclear for how long these patients should be followed up. To address this issue, we did conditional survival modelling using the International Watch & Wait Database (IWWD), which is a large-scale registry of patients with a clinical complete response after neoadjuvant chemotherapy who have been managed by a watch-and-wait strategy. We did a retrospective, multicentre registry study using a dataset from the IWWD, which includes data from 47 clinics across 15 countries. We selected patients (aged ≥18 years) with rectal cancer who had a clinical complete response after neoadjuvant chemotherapy, and who were subsequently managed by a watch-and-wait strategy between Nov 25, 1991, and Dec 31, 2015. Patients who had not achieved a clinical complete response or who had undergone any surgical procedure were excluded. The criteria used for defining a clinical complete response and the specific surveillance strategies were at the discretion of each participating centre. We used conditional survival modelling to estimate the probability of patients remaining free of local regrowth or distant metastasis for an additional 2 years after sustaining a clinical complete response or being distant metastasis-free for 1, 3, and 5 years from the date of the decision to commence watch and wait. The primary outcomes were conditional local regrowth-free survival at 3 years, and conditional distant metastasis-free survival at 5 years. We identified 793 patients in the IWWD with clinical complete response who had been managed by a watch-and-wait strategy. Median follow-up was 55·2 months (IQR 36·0-75·6). The probability of remaining free from local regrowth for an additional 2 years if a patient had a sustained clinical complete response for 1 year was 88·1% (95% CI 85·8-90·9), for 3 years was 97·3% (95·2-98·6), and for 5 years was 98·6% (97·6-100·0). The probably of remaining free from distant metastasis for a further 2 years in patients who had a clinical complete response without distant metastasis for 1 year was 93·8% (92·3-95·9), for 3 years was 97·8% (96·6-99·3), and for 5 years was 96·6% (94·0-98·9). These results suggest that the intensity of active surveillance in patients with rectal cancer managed by a watch-and-wait approach could be reduced if they achieve and maintain a clinical complete response within the first 3 years of starting this approach. European Registration of Cancer Care, financed by the European Society of Surgical Oncology, the Champalimaud Foundation Lisbon, the Bas Mulder Award, granted by the Alpe d'HuZes Foundation and the Dutch Cancer Society, the European Research Council Advanced Grant, and the National Institute of Health and Research Manchester Biomedical Research Centre.


Herbal bioactivation: the good, the bad and the ugly.
It has been well established that the formation of reactive metabolites of drugs is associated with drug toxicity. Similarly, there are accumulating data suggesting the role of the formation of reactive metabolites/intermediates through bioactivation in herbal toxicity and carcinogenicity. It has been hypothesized that the resultant reactive metabolites following herbal bioactivation covalently bind to cellular proteins and DNA, leading to toxicity via multiple mechanisms such as direct cytotoxicity, oncogene activation, and hypersensitivity reactions. This is exemplified by aristolochic acids present in Aristolochia spp, undergoing reduction of the nitro group by hepatic cytochrome P450 (CYP1A1/2) or peroxidases in extrahepatic tissues to reactive cyclic nitrenium ion. The latter was capable of reacting with DNA and proteins, resulting in activation of H-ras oncogene, gene mutation and finally carcinogenesis. Other examples are pulegone present in essential oils from many mint species; and teucrin A, a diterpenoid found in germander (Teuchrium chamaedrys) used as an adjuvant to slimming diets. Extensive pulegone metabolism generated p-cresol that was a glutathione depletory, and the furan ring of the diterpenoids in germander was oxidized by CYP3A4 to reactive epoxide which reacts with proteins such as CYP3A and epoxide hydrolase. On the other hand, some herbal/dietary constituents were shown to form reactive intermediates capable of irreversibly inhibiting various CYPs. The resultant metabolites lead to CYP inactivation by chemical modification of the heme, the apoprotein, or both as a result of covalent binding of modified heme to the apoprotein. Some examples include bergamottin, a furanocoumarin of grapefruit juice; capsaicin from chili peppers; glabridin, an isoflavan from licorice root; isothiocyanates found in all cruciferous vegetables; oleuropein rich in olive oil; dially sulfone found in garlic; and resveratrol, a constituent of red wine. CYPs have been known to metabolize more than 95% therapeutic drugs and activate a number of procarcinogens as well. Therefore, mechanism-based inhibition of CYPs may provide an explanation for some reported herb-drug interactions and chemopreventive activity of herbs. Due to the wide use and easy availability of herbal medicines, there is increasing concern about herbal toxicity. The safety and quality of herbal medicine should be ensured through greater research, pharmacovigilance, greater regulatory control and better communication between patients and health professionals.


Epstein-Barr Virus (EBV): infection, propagation, quantitation, and storage.
Epstein-Barr virus (EBV) was first reported as the etiological agent of Burkitt's lymphoma in 1964. Since then, EBV has also been associated with nasopharyngeal carcinoma, which is highly prevalent in Southeast Asia, as well as infectious mononucleosis, complications of AIDS, and transplant-related B cell lymphomas. This virus has further been linked with T cell lymphomas and Hodgkin's disease, establishing the concept of a wide spectrum of EBV-associated malignant disorders. So far, there are a number of EBV-infected cell lines established that can be induced for production of infectious viral progeny and that facilitate the study of the mechanism of EBV-related infection, transformation, and oncogenesis. This unit describes procedures for the preparation of EBV virion particles and in vitro infection of cells with EBV. In addition, procedures for quantitation and storage of the virus are provided.


Data publication consensus and controversies.
The movement to bring datasets into the scholarly record as first class research products (validated, preserved, cited, and credited) has been inching forward for some time, but now the pace is quickening. As data publication venues proliferate, significant debate continues over formats, processes, and terminology. Here, we present an overview of data publication initiatives underway and the current conversation, highlighting points of consensus and issues still in contention. Data publication implementations differ in a variety of factors, including the kind of documentation, the location of the documentation relative to the data, and how the data is validated. Publishers may present the data as supplemental material to a journal article, with a descriptive "data paper," or independently. Complicating the situation, different initiatives and communities use the same terms to refer distinct but overlapping concepts. For instance, the term "published" means that the data is publicly available and citable to virtually everyone, but it may or may not imply that the data has been peer-reviewed. In turn, what is meant by data peer review is far from defined; standards and processes encompass the full range employed in reviewing the literature, plus some novel variations. Basic data citation is a point of consensus, but the general agreement on the core elements of a dataset citation frays if the data is dynamic or part of a larger set. Even as data publication is being defined, some are looking past publication to other metaphors, notably "data as software," for solutions to the more stubborn problems.


Updating a systematic review--what difference did it make? Case study of nicotine replacement therapy.
To examine the effect of updating a systematic review of nicotine replacement therapy on its contents and conclusions. We examined the effects of regular updating of a systematic review of nicotine replacement therapy for smoking cessation. We considered two outcomes. First, we assessed the effect of adding new data to meta-analyses, comparing results in 2000 with the results in 1994. Second, we assessed qualitatively the ways in which the nature of the questions addressed by the review had changed between the two dates. For the first outcome, we compared the number of trials, the pooled estimate of effect using the odds ratio, and the results of pre-specified subgroup analyses, for nicotine gum and patch separately. Using a test for interaction, we assessed whether differences between estimates were statistically significant. There were ten new trials of nicotine gum between 1994 and 2000, and the meta-analytic effect changed little. For the nicotine patch the number of trials increased from 9 to 30, and the meta-analytic effect fell from 2.07 (95% CI 1.64 - 2.62) to 1.73 (95% CI 1.56 - 1.93). Apparent differences in relative effect in sub-groups found in 1994 were not found in 2000. The updated systematic review addressed a number of questions not identified in the original version. Updating the meta-analyses lead to a more precise estimate of the likely effect of the nicotine patch, but the clinical message was unchanged. Further placebo controlled NRT trials are not likely to add to the evidence base. It is questionable whether updating the meta-analyses to include them is worthwhile. The content of the systematic review has, however, changed, with the addition of data addressing questions not considered in the original review. There is a tension between the principle of identifying the important questions prior to conducting a review, and keeping the review up to date as primary research identifies new avenues of enquiry.


Tetronarce cowleyi, sp. nov., a new species of electric ray from southern Africa (Chondrichthyes: Torpediniformes: Torpedinidae).
A new species of torpedo ray, Tetronarce cowleyi, sp. nov., is described from specimens collected from the southeastern Atlantic Ocean. The new species is placed in the genus Tetronarce based on a uniform dorsal coloration and absence of papillae around the spiracles. The new species is distinguished from its closest congeners, the North Atlantic Tetronarce nobiliana Bonnaparte, 1835, and southwestern Atlantic Tetronarce puelcha Lahille, 1926, by a combination of morphological characteristics including a shorter spiracular length, a proportionally greater head length as measured between snout margin and fifth gill openings, a proportionally greater preoral snout length, a uniform shiny black or dark gray dorsal surface, lacking any prominent markings, and a creamy white ventral color with dark edges in juveniles but fading with growth. Teteronarce cowleyi, sp. nov., is further distinguished from T. nobiliana by its more circular anterior disc shape (vs. relatively straight in T. nobiliana), fewer tooth rows (32/28 vs. 38-53/38-52 in T. nobiliana), greater mouth width (1.5-1.7 times as great as interorbital width vs. 0.5-0.6 times interorbital width in T. nobiliana), smaller distance between second dorsal and caudal fins (3.5-4.9% vs. 6.6-6.8% in T. nobiliana), and a clasper length extending nearly to lower caudal fin origin (claspers in T. nobiliana that extend only two-thirds distance between second dorsal and caudal fins). Teteronarce cowleyi, sp. nov., is known from Walvis Bay, Namibia to Algoa Bay, Eastern Cape, South Africa, at depths of 110 to 457 m.


Metastable electronic states in uranium tetrafluoride.
The DFT+U approach, where U is the Hubbard-like on-site Coulomb interaction, has successfully been used to improve the description of transition metal oxides and other highly correlated systems, including actinides. The secret of the DFT+U approach is the breaking of d or f shell orbital degeneracy and adding an additional energetic penalty to non-integer occupation of orbitals. A prototypical test case, UO2, benefits from the +U approach whereby the bare LDA method predicts UO2 to be a ferromagnetic metal, whereas LDA+U correctly predicts UO2 to be insulating. However, the concavity of the energetic penalty in the DFT+U approach can lead to a number of convergent "metastable" electronic configurations residing above the ground state. Uranium tetrafluoride (UF4) represents a more complex analogy to UO2 in that the crystal field has lower symmetry and the unit cell contains two symmetrically distinct U atoms. We explore the metastable states in UF4 using several different methods of selecting initial orbital occupations. Two methods, a "pre-relaxation" method wherein an initial set of orbital eigenvectors is selected via the self-consistency procedure and a crystal rotation method wherein the x, y, z axes are brought into alignment with the crystal field, are explored. We show that in the case of UF4, which has non-collinearity between its crystal axes and the U atoms' crystal field potentials, the orbital occupation matrices are much more complex and should be analyzed using a novel approach. In addition to demonstrating a complex landscape of metastable electronic states, UF4 also shows significant hybridization in U-F bonding, which involves non-trivial contributions from s, p, d, and f orbitals.


Anchoring effect in legal decision-making: A meta-analysis.
Objective: We conducted a meta-analysis to examine whether numeric decision-making in law is susceptible to the effect of (possibly arbitrary) values present in the decision contexts (anchoring effect) and to investigate which factors might moderate this effect. Hypotheses: We predicted that the presence of numeric anchors would bias legal decision-makers' judgment in the direction of the anchor value. We hypothesized that the effect size of anchoring would be moderated by several variables, which we grouped into three categories: methodological (type of stimuli; type of sample), psychological (standard vs. basic paradigm; anchor value; type of scale on which the participants assessed the target value), and legal (relevance of the anchor; type of the anchor; area of law to which the presented case belonged; presence of any salient numeric values other than the main anchor). Method: Twenty-nine studies (93 effect sizes; N = 8,549) met the inclusion criteria. We divided them into two groups, depending on whether they included a control group, and calculated the overall effect size using a random-effects Model with robust variance estimation. We assessed the influence of moderators using random effects metaregression. Results: The overall effect sizes of anchoring for studies with a control group (z = .27, 95% CI [.21, .33], d = .58, 95% CI [.44, .73]) and without a control group (z = .39, 95% CI [.31, .47], d = .91, 95% CI [.69, 1.12]) were both significant, although we provide some evidence of possible publication bias. We found preliminary evidence of a potential moderating effect of some legally relevant factors, such as legal expertise or the anchor relevance. Conclusions: Existing research indicates anchoring effects exist in legal contexts. The influence of anchors seems to depend on some situational factors, which paves the way for future research on countering the problematic effect in legal settings. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Modeling Endogenous Mobility in Earnings Determination.
We evaluate the bias from endogenous job mobility in fixed-effects estimates of worker- and firm-specific earnings heterogeneity using longitudinally linked employer-employee data from the LEHD infrastructure file system of the U.S. Census Bureau. First, we propose two new residual diagnostic tests of the assumption that mobility is exogenous to unmodeled determinants of earnings. Both tests reject exogenous mobility. We relax exogenous mobility by modeling the matched data as an evolving bipartite graph using a Bayesian latent-type framework. Our results suggest that allowing endogenous mobility increases the variation in earnings explained by individual heterogeneity and reduces the proportion due to employer and match effects. To assess external validity, we match our estimates of the wage components to out-of-sample estimates of revenue per worker. The mobility-bias corrected estimates attribute much more of the variation in revenue per worker to variation in match quality and worker quality than the uncorrected estimates.


Origin and evolutionary process of the CNS elucidated by comparative genomics analysis of planarian ESTs.
Among the bilateral animals, a centralized nervous system is found in both the deuterostome and protostome. To address the question of whether the CNS was derived from a common ancestor of deuterostomes and protostomes, it is essential to know kinds of genes existed in the CNS of the putative common ancestor and to trace the evolutionary divergence of genes expressed in the CNS. To answer these questions, we took a comparative approach using different species, particularly focusing on one of the lower bilateral animals, the planarian (Platyhelminthes, Tricladida), which is known to possess a CNS. We determined the nucleotide sequence of ESTs from the head portion of planarians, obtaining 3,101 nonredundant EST clones. As a result of homology searches, we found that 116 clones had significant similarity to known genes related to the nervous system. Here, we compared these 116 planarian EST clones with all ORFs of the complete genome sequences of the human, fruit fly, and nematode, and showed that >95% of these 116 nervous system-related genes, including genes involved in brain or neural morphogenesis, were commonly shared among these organisms, thus providing evidence at the molecular level for the existence of a common ancestral CNS. Interestingly, we found that approximately 30% of planarian nervous system-related genes had homologous sequences in Arabidopsis and yeast, which do not possess a nervous system. This implies that the origin of nervous system-related genes greatly predated the emergence of the nervous system, and that these genes might have been recruited toward the nervous system.


Changing urban phosphorus metabolism: Evidence from Longyan City, China.
Rapid worldwide urbanization calls for a better understanding of phosphorus (P) metabolism and the interaction of the physical, ecological and social drivers of P cycling in urban systems. We quantified the P metabolism in Longyan, a city with a major agricultural economy, and analyzed its long-term trends over the rapid urbanization period of 1985-2010. Both input P (from 4811 t P to 14,296 t P) and output P (from 4565 t P to 13,509 t P) increased significantly. The agricultural subsystem contributed most to the P metabolism, accounting for 85% of total P input. The share of P input lost to the environment, i.e. discharge to water, accumulation in the soil and landfill, increased from 66% to 72%, while food production efficiency decreased from 48% to 29%. Per capita P input showed linear relationships with the Human Development Index (HDI), S-curve relationship with the urbanization rate, and logistic curve relationship with per capita disposable income. A more meat-based diet shift both in Longyan and surrounding cities greatly affected Longyan's food production structure. Our results demonstrate that P metabolic quantity, configuration, and efficiency in production systems can change drastically in response to changes in consumer and producer behavior as well as in socioeconomic structure. A larger regional scale should be considered in urban P management, when trying to mitigate the increase in P use. The results also imply that sustainable urban P management will require a system-wide, cross-sector and cross-boundary approach.


Intrinsic MANOVA for Riemannian manifolds with an application to Kendall's space of planar shapes.
We propose an intrinsic multifactorial model for data on Riemannian manifolds that typically occur in the statistical analysis of shape. Due to the lack of a linear structure, linear models cannot be defined in general; to date only one-way MANOVA is available. For a general multifactorial model, we assume that variation not explained by the model is concentrated near elements defining the effects. By determining the asymptotic distributions of respective sample covariances under parallel transport, we show that they can be compared by standard MANOVA. Often in applications manifolds are only implicitly given as quotients, where the bottom space parallel transport can be expressed through a differential equation. For Kendall's space of planar shapes, we provide an explicit solution. We illustrate our method by an intrinsic two-way MANOVA for a set of leaf shapes. While biologists can identify genotype effects by sight, we can detect height effects that are otherwise not identifiable.


An astonishing journey into reproductive genetics since the 1950's.
Training in genetics in Edinburgh in the 1950s led to a PhD on the developmental biology of mouse embryos with unusual chromosomal complements. Fundamental aspects of reproduction under study included ovulation induction, oocyte maturation and embryonic growth to blastocysts. It led to the introduction of embryo stem cells, preimplantation genetic diagnosis, the exact timing of human oocyte maturation in vitro and studies on fertilising human eggs in vitro to alleviate human infertility. My work was helped by studies on sperm capacitation and the physiology of fertilization in domestic and laboratory species by Thibault, Dauzier, Austin, Chang, Yanagimachi and others. I met Charles Thibault at a meeting in Cambridge U.K. where he criticised the work of Moricard, and then frequently on lecture circuits. Impressed by his grandeur but not his doubts about human IVF, Steptoe and I initiated human embryo transfers and the birth of Louise Brown. Details of her pregnancy had to be confidential to reduce the risks of abortion associated with the intrusion of numerous newsmen chasing the story. I was compelled to withold this information at a meeting in Paris in the late 1960s when I had to leave early to return to UK. This omission annoyed Thibault and led to our celebrated quarrel. I felt he failed to appreciate the complexity, the implications of this pregnancy and an astonishing future. So much was at stake, including IVF, stem cells and preimplantation diagnosis to help millions of patients. Some months later, our dispute was ended even if somewhat formally. Nevertheless it is a pleasure to recall how we shared so much in common. I still admire him as an inspiration to many colleagues and students, and a father figure in French agricultural research.


Presentation, antibiotic management and associated outcome in Polish adults presenting with acute cough/LRTI.
In-depth knowledge of existing practice is required to inform interventions aimed at antibiotic prescribing quality improvement. We set out to describe the presentation, antimicrobial management and associated outcome of adults presenting in general practice with acute cough/lower respiratory tract infection (LRTI) in Poland. Observational study of 301 adults with acute cough/LRTI. Clinicians completed a case report form (CRF) describing presentation, history and management and patients completed a symptom diary for up to 28 days after consultation. Two hundred and twenty-one patients (with CRF and symptom diary completed) were analysed. The median duration of feeling unwell before presentation was 4 days. Clinicians recorded an average of eight symptoms for patients at presentation. Apart from cough, patients most commonly reported feeling generally unwell (91.9%), limitation of normal activities (80.5%), coryza (80.1%) and phlegm production (76.0%). Auscultation abnormalities were present in 55.0%. Overall, medicines were prescribed for 95.0%; 72.4% were prescribed antibiotics [mostly macrolides/lincosamides (38.8%) and amoxicillin/co-amoxiclav (36.3%)) with 11.3% advised to take antibiotics only if still necessary after a specified delay. Mucolytics were prescribed for 61.1%. Antibiotic prescription was strongly associated with a diagnosis of LRTI and the presence of auscultation abnormalities. The median duration of cough after presentation was 8 days. Antibiotics continue to be frequently prescribed for acute cough/LRTI in Poland, and the decision to prescribe was strongly associated with clinicians' findings of abnormalities on auscultation and diagnosis of LRTI. Delayed prescribing was infrequent. Mucolytics were commonly prescribed despite evidence of no effect.


Interpersonal distance and social anxiety in autistic spectrum disorders: A behavioral and ERP study.
An inherent feature of social interactions is the use of social space or interpersonal distance-the space between one individual and another. Because social deficits are core symptoms of Autistic Spectrum Disorder (ASD), we hypothesized that individuals on this spectrum will exhibit abnormal interpersonal distance preferences. The literature on interpersonal distance in ASD is not conclusive. While some studies show preferences for closer distances among this group, others show preferences for farther distances than controls. A common symptom of ASD that may explain the variance in responses to interpersonal distance in this population is social anxiety (SA), which has been shown to correlate with interpersonal distance preferences. In the current study, we investigated interpersonal distance preferences in a group of individuals with ASD using both behavioral and ERP measures. We found greater variance in interpersonal distance preferences in the ASD group than in the control group. Furthermore, we showed that this variance can be explained by differences in SA level and can be predicted by the N1 amplitude, an early ERP component related to attention and discrimination processes. These results hint at the early sensory and attentional processes that may be affecting higher social behaviors, both in subclinical and in clinical populations.


Cardiovascular parameter changes in patients with erectile dysfunction using pde-5 inhibitors: a study with sildenafil and vardenafil.
Sildenafil is the most prescribed oral agent for patients with erectile dysfunction (ED). Vardenafil is a new phosphodiesterase type 5 (Pde-5) inhibitor that was approved by the US Food and Drug Administration last year to treat patients with ED of various causes. Both of these Pde-5 inhibitors have vasodilating properties and effects on blood pressure (BP), and like nitrates, they work through the nitric oxide cyclic guanosine monophosphate pathway. The aim of this study was to investigate the influence of these Pde-5 inhibitors on BP and heart rate (HR) in normotensive men with ED by a crossover comparison. Thirty-five patients with ED were enrolled to evaluate and compare the effect of sildenafil (50 mg) and vardenafil (10 mg) on BP and HR. At the screening (baseline [B]) visit, sitting systolic blood pressure (B-SBP), diastolic blood pressure (B-DBP), and HR were measured. We performed a multiple administration for both drugs and, therefore, multiple measurements of BP and HR changes, 3 doses a week, on alternate days, late in the afternoon, and on an empty stomach. B-SBP, B-DBP, and HR were recorded before each 50-mg sildenafil dosing and after 30, 60, 120, and 240 minutes. Data were averaged over the 4 time points and compared with the baseline values obtained before each dosing. After a 3-week wash-out period, patients were crossed over to vardenafil (10 mg) with the same study design. After administration of both drugs, we observed a statistically significant decrease of BP and an increase of HR. On average, sildenafil caused a decrease of SBP ranging from 5.1 +/- 3.9 mm Hg during the first dosing to 4.7 +/- 4.2 mm Hg during the third dosing, DBP ranged from 4.4 +/- 4.9 to 4 +/- 4.1 mm Hg, and HR increased 1.8 +/- 2.0 bpm (first dose) and 1.2 +/- 0.9 bpm (third dose). With vardenafil, we recorded a greater variation for SBP and DBP. SBP decreased from 8.02 +/- 8.0 mm Hg during the first dosing to 5.4 +/- 5.5 mm Hg during the third dosing, whereas DBP decreased from 6.6 +/- 7.2 to 5.0 +/- 5.3 mm Hg, respectively. Recorded HR showed an increase of 3.1 +/- 3.2 bpm (first dose) and 2.4 +/- 2.3 bpm (third dose). After the first vardenafil administration, we recorded fainting episodes in 3 patients because of a decrease in BP greater than 20 mm Hg. Two of the patients were in therapy with doxazosin for benign prostatic hyperplasia (BPH). Cardiovascular response was not significantly different after the first dose between the 2 treatments. Vardenafil demonstrated clinically significant differences (fainting) with respect to sildenafil only during the first doses. We suggest that before starting therapies with Pde-5 inhibitors, particularly with the newer ones, that baseline cardiovascular parameters are measured and monitored, especially during the first dose, because of the presence of a "first dose effect." Moreover, it is necessary to pay particular attention to those patients in treatment with other drugs that could have a synergistic hypotensive effect as a result of vasodilation potentiation.


Can primary care physicians' questions be answered using the medical journal literature?
Medical librarians and informatics professionals believe the medical journal literature can be useful in clinical practice, but evidence suggests that practicing physicians do not share this belief. The authors designed a study to determine whether a random sample of "native" questions asked by primary care practitioners could be answered using the journal literature. Participants included forty-nine active, nonacademic primary care physicians providing ambulatory care in rural and nonrural Oregon, and seven medical librarians. The study was conducted in three stages: (1) office interviews with physicians to record clinical questions; (2) online searches to locate answers to selected questions; and (3) clinician feedback regarding the relevance and usefulness of the information retrieved. Of 295 questions recorded during forty-nine interviews, 60 questions were selected at random for searches. The average total time spent searching for and selecting articles for each question was forty-three minutes. The average cost per question searched was $27.37. Clinician feedback was received for 48 of 56 questions (four physicians could not be located, so their questions were not used in tabulating the results). For 28 questions (56%), clinicians judged the material relevant; for 22 questions (46%) the information provided a "clear answer" to their question. They expected the information would have had an impact on their patient in nineteen (40%) cases, and an impact on themselves or their practice in twenty-four (51%) cases. If the results can be generalized, and if the time and cost of performing searches can be reduced, increased use of the journal literature could significantly improve the extent to which primary care physicians' information needs are met.


How calcium signals in myocytes and pericytes are integrated across in situ microvascular networks and control microvascular tone.
The microcirculation is the site of gas and nutrient exchange. Control of central or local signals acting on the myocytes, pericytes and endothelial cells within it, is essential for health. Due to technical problems of accessibility, the mechanisms controlling Ca2+ signalling and contractility of myocytes and pericytes in different sections of microvascular networks in situ have not been investigated. We aimed to investigate Ca2+ signalling and functional responses, in a microcirculatory network in situ. Using live confocal imaging of ureteric microvascular networks, we have studied the architecture, morphology, Ca2+ signalling and contractility of myocytes and pericytes. Ca2+ signals vary between distributing arcade and downstream transverse and precapillary arterioles, are modified by agonists, with sympathetic agonists being ineffective beyond transverse arterioles. In myocytes and pericytes, Ca2+ signals arise from Ca2+ release from the sarcoplasmic reticulum through inositol 1,4,5-trisphosphate-induced Ca2+ release and not via ryanodine receptors or Ca2+ entry into the cell. The responses in pericytes are less oscillatory, slower and longer-lasting than those in myocytes. Myocytes and pericytes are electrically coupled, transmitting Ca2+ signals between arteriolar and venular networks dependent on gap junctions and Ca2+ entry via L-type Ca2+ channels. Endothelial Ca2+ signalling inhibits intracellular Ca2+ oscillations in myocytes and pericytes via L-arginine/nitric oxide pathway and intercellular propagating Ca2+ signals via EDHF. Increases of Ca2+ in pericytes and myocytes constrict all vessels except capillaries. These data reveal the structural and signalling specializations allowing blood flow to be regulated by myocytes and pericytes.


Application of the Abel integral equation to spectrographic data.
An improved method is proposed for applying the Abel integral equation to radiance data gathered from rotationally symmetric sources by side-on observation. The method involves dividing the data into a number of segments going out from the center. A least-squares polynomial is then fitted to each segment so that the inverted Abel integral equation can be integrated exactly to yield the emission coefficient. It is shown, with test curves approximating spectral line shapes from nonhomogeneous sources, that the method becomes superior to the other more common numerical methods of reducing such data as the number of data points becomes large. This superiority increases as the data scatter increases.


Asexual but Not Clonal: Evolutionary Processes in Automictic Populations.
Many parthenogenetically reproducing animals produce offspring not clonally but through different mechanisms collectively referred to as automixis. Here, meiosis proceeds normally but is followed by a fusion of meiotic products that restores diploidy. This mechanism typically leads to a reduction in heterozygosity among the offspring compared to the mother. Following a derivation of the rate at which heterozygosity is lost at one and two loci, depending on the number of crossovers between loci and centromere, a number of models are developed to gain a better understanding of basic evolutionary processes in automictic populations. Analytical results are obtained for the expected neutral genetic variation, effective population size, mutation-selection balance, selection with overdominance, the spread of beneficial mutations, and selection on crossover rates. These results are complemented by numerical investigations elucidating how associative overdominance (two off-phase deleterious mutations at linked loci behaving like an overdominant locus) can in some cases maintain heterozygosity for prolonged times, and how clonal interference affects adaptation in automictic populations. These results suggest that although automictic populations are expected to suffer from the lack of gene shuffling with other individuals, they are nevertheless, in some respects, superior to both clonal and outbreeding sexual populations in the way they respond to beneficial and deleterious mutations. Implications for related genetic systems such as intratetrad mating, clonal reproduction, selfing, as well as different forms of mixed sexual and automictic reproduction are discussed.


Effects of deep brain stimulation of the subthalamic nucleus on freezing of gait in Parkinson's disease: a prospective controlled study.
Freezing of gait (FOG) is a debilitating gait disorder in Parkinson's disease (PD) with partial responsiveness to dopaminergic medication. To date, notions about the effects of subthalamic deep brain stimulation (STN-DBS) on FOG remain controversial. To compare the effects of bilateral STN-DBS and continued best medical treatment (BMT) on FOG occurrence, FOG severity and clinical outcomes in PD patients at 6 and 12 months follow-up. In this prospective, controlled study, 41 PD patients with at least 5 years disease duration participated. Twenty-four subjects (20 with FOG) were treated with STN-DBS and seventeen (15 with FOG) continued BMT. The primary outcome was the New Freezing of Gait Questionnaire (NFOGQ) at 6 months postsurgery. Other outcomes were the NFOGQ at 12 months and clinical outcomes (Unified Parkinson's Disease Rating Scale III (UPDRS III), timed gait, falls and quality of life) at both time points. STN-DBS increased the likelihood to convert from being a freezer to a non-freezer at 6 and 12 months follow-up (relative risk reduction=0.4). However, 45% of baseline freezers still experienced FOG 6 and 12 months postsurgery although with reduced severity. Three baseline non-freezers (1/2 BMT-treated, 2/4 STN-DBS-treated) developed FOG during follow-up. STN-DBS-induced benefits on FOG were mostly mediated by baseline levodopa equivalent dose, altered medication-intake and reduced motor fluctuations. In contrast to continued BMT, STN-DBS reduced FOG occurrence and severity at 6 months postsurgery with largely sustained effects at 12 months follow-up. Longer follow-up periods are needed to test whether FOG improvements after STN-DBS persist with disease progression.