Comparison of irrigation to floss as an adjunct to tooth brushing: effect on bleeding, gingivitis, and supragingival plaque.
The purpose of this twenty-eight day, randomized, single-blind clinical trial was to assess the efficacy of the addition of daily oral irrigation to both power and manual tooth brushing, compared to a traditional regimen of manual tooth brushing and flossing, to determine which regimen had the greatest effect on the reduction of gingival bleeding, gingivitis, and supragingival plaque. The study was designed for a total of 105 subjects to participate in a twenty-eight day trial, with 35 subjects randomly assigned to one of three groups: Group 1-manual toothbrush and floss; Group 2-manual toothbrush and dental water jet; and Group 3-sonic toothbrush and dental water jet. All subjects received written and verbal instructions for using their regimens. Subjects were asked to brush for a timed two minutes, twice per day, with the brush and the dentifrice provided, and to refrain from using any additional oral hygiene aids. Subjects using the dental water jet were instructed to use the water jet on a medium setting, irrigating once per day with 500 ml of luke warm water. Subjects using the dental floss were instructed to use the floss once daily. Subjects were examined by two calibrated examiners, and data were collected at baseline (BSL), 14 days (D14), and 28 days (D28). Subjects were asked to abstain from any oral hygiene for 12 hours prior to each study visit. Subjects were scored using the Carter and Barnes Bleeding Index, Löe and Silness Gingival Index, and the Proximal/Marginal Plaque Index. Mean scores on the three indices for the three groups were used for statistical analysis at each time point. Additionally, the means were used for comparisons as change from baseline and percent change from baseline at D14 and D28. The significance of percentage change in each index from baseline to D14 and D28 was evaluated using a one-tailed t-test. Significant differences are reported at alpha < or = 0.05 for these planned group comparisons. Thirty-one subjects in Group 1, 32 subjects in Group 2, and 32 subjects in Group 3 completed the study. Bleeding Index: Groups 2 and 3, the irrigation groups, were statistically significantly more effective than Group 1 in reducing the bleeding index at D14 and D28, whether measured by mean reduction or percentage reduction. Gingival Index: At D14, both irrigation groups demonstrated a statistically significantly greater reduction in the gingival index compared to brushing and flossing for the facial surfaces. There was no significant difference between groups for the lingual surface at D14. At D28 there was a significant difference between Groups 1 and 2 for both the facial and lingual surfaces. Plaque Index: There was one significant difference between groups for the plaque index measured on the lingual surfaces. The manual toothbrush and floss were less effective than the sonic toothbrush and irrigation. Group 3 was also significantly better than Group 1 in reducing the plaque index on the facial surfaces at both D14 and D28. On plaque percentage reduction on the facial surface, Group 2 was significantly better than Group 1 at D14. There was no statistical difference between Group 1 and Group 2 at D28. The results of this clinical trial indicate that when combined with manual or sonic tooth brushing, oral irrigation is an effective alternative to manual tooth brushing and dental floss for reducing bleeding, gingival inflammation, and plaque removal.


Female-to-male patients have high prevalence of unsatisfactory Paps compared to non-transgender females: implications for cervical cancer screening.
Little is known about whether and how screening for cancers of natal reproductive structures, including cervical cancer, in female-to-male (FTM) transgender individuals differs from cancer screening among non-transgender females. To investigate anecdotal reports from clinicians of high rates of inadequate Papanicolaou (Pap) tests among transgender men. Results of Pap tests performed on 233 FTM and 3,625 female patients at an urban community health center between 2006 and 2012 were extracted from an electronic medical record. Compared to female patients, FTM patients were more likely to have an inadequate Pap, with prevalence of inadequate samples 8.3 times higher among tests of FTM patients (10.8% vs. 1.3% of tests). FTM patients had over ten times higher odds of having an inadequate Pap after adjusting for age, race, and body mass index (AOR = 10.77, 95% CI = 6.83, 16.83). When years on testosterone therapy was added to the model, the relationship between transgender identity and Pap inadequacy was attenuated, but remained strongly associated (AOR = 6.01, 95% CI = 3.00, 11.50), and time on testosterone was also associated (AOR = 1.19, 95% CI 1.04, 1.36). FTM patients were more likely than females to have had multiple inadequate tests, and had longer latency to follow-up testing. The high unsatisfactory sample prevalence among FTM patients is likely due to a combination of physical changes induced by testosterone therapy and provider/patient discomfort with the exam. Clinicians should receive training in increasing comfort for FTM patients during the exam. FTM patients should be alerted that high rates of inadequate screening may require follow-up testing. Alternatives to repeated Pap testing, such as cytologic reprocessing of inadequate samples or primary human papillomavirus (HPV) DNA screening, should be studied for efficacy and acceptability among FTM patients.


Divergent phenological response to hydroclimate variability in forested mountain watersheds.
Mountain watersheds are primary sources of freshwater, carbon sequestration, and other ecosystem services. There is significant interest in the effects of climate change and variability on these processes over short to long time scales. Much of the impact of hydroclimate variability in forest ecosystems is manifested in vegetation dynamics in space and time. In steep terrain, leaf phenology responds to topoclimate in complex ways, and can produce specific and measurable shifts in landscape forest patterns. The onset of spring is usually delayed at a specific rate with increasing elevation (often called Hopkins' Law; Hopkins, 1918), reflecting the dominant controls of temperature on greenup timing. Contrary with greenup, leaf senescence shows inconsistent trends along elevation gradients. Here, we present mechanisms and an explanation for this variability and its significance for ecosystem patterns and services in response to climate. We use moderate-resolution imaging spectro-radiometer (MODIS) Normalized Difference Vegetation Index (NDVI) data to derive landscape-induced phenological patterns over topoclimate gradients in a humid temperate broadleaf forest in southern Appalachians. These phenological patterns are validated with different sets of field observations. Our data demonstrate that divergent behavior of leaf senescence with elevation is closely related to late growing season hydroclimate variability in temperature and water balance patterns. Specifically, a drier late growing season is associated with earlier leaf senescence at low elevation than at middle elevation. The effect of drought stress on vegetation senescence timing also leads to tighter coupling between growing season length and ecosystem water use estimated from observed precipitation and runoff generation. This study indicates increased late growing season drought may be leading to divergent ecosystem response between high and low elevation forests. Landscape-induced phenological patterns are easily observed over wide areas and may be used as a unique diagnostic for sources of ecosystem vulnerability and sensitivity to hydroclimate change.


Three-year clinical performance of a HEMA-free one-step self-etch adhesive in non-carious cervical lesions.
Despite the fact that one-step adhesives are currently used routinely in clinical practice, long-term studies on their clinical performance are scarce. The objective of this randomized controlled clinical trial was to test the hypothesis that a 2-hydroxyethyl methacrylate (HEMA)-free one-step self-etch adhesive performs worse than a conventional multistep etch-and-rinse adhesive. Two-hundred and seventy-six non-carious cervical lesions in 52 patients were restored with a micro-hybrid composite (Gradia Direct; GC). These restorations were bonded in random order either with the HEMA-free one-step adhesive G-Bond (GC) or with the 'gold-standard' (control) three-step adhesive Optibond FL (Kerr). The restorations were evaluated after 6, 12, 24, and 36 months of clinical service regarding retention, marginal adaptation, microleakage, caries occurrence, and sensitivity. After a medium-long period of 3 yr, similar success in clinical performance was observed for the simplified all-in-one adhesive and the conventional three-step adhesive. However, the one-step adhesive exhibited significantly more incisal marginal defects and discolorations. Whereas marginal degradation appeared to arrest for the multistep etch-and-rinse adhesive after 12 months, the enamel margins of the restorations bonded with the one-step self-etch adhesive continued to deteriorate. These incisal marginal defects were, however, small and could easily be removed by polishing. For both adhesives, large and sclerosed lesions appeared to be at higher risk of retention loss.


The asylum, the Poor Law, and a reassessment of the four-shilling grant: admissions to the county asylums of Yorkshire in the nineteenth century.
In the mid-1870s, the British government introduced a grant that transferred a proportion of the cost of asylum care from local to central funds. Typically, this has been seen by contemporary and more recent commentators as part of the explanation for the therapeutic failure of the County and Borough Asylums, and for their degeneration into custodial institutions. Building on recent work on the Poor Law, the aim of this article is to reassess the impact of the grant using both quantitative and qualitative evidence. Contrasting the records of two County Asylums with the annual reports of the Lunacy Commissioners, it shows that there is little evidence to suggest that the grant was responsible for a change in either the size or composition of the asylum population. Ultimately, it argues that the admission of patients in general, and the admission and discharge of chronic cases in particular, rested with longer-term factors than simply the introduction of one fiscal incentive.


Simultaneous imaging of amyloid-β and lipids in brain tissue using antibody-coupled liposomes and time-of-flight secondary ion mass spectrometry.
The spatial localization of amyloid-β peptide deposits, the major component of senile plaques in Alzheimer's disease (AD), was mapped in transgenic AD mouse brains using time-of-flight secondary ion mass spectrometry (ToF-SIMS), simultaneously with several endogenous molecules that cannot be mapped using conventional immunohistochemistry imaging, including phospholipids, cholesterol and sulfatides. Whereas the endogenous lipids were detected directly, the amyloid-β deposits, which cannot be detected as intact entities with ToF-SIMS because of extensive ion-induced fragmentation, were identified by specific binding of deuterated liposomes to antibodies directed against amyloid-β. Comparative investigation of the amyloid-β deposits using conventional immunohistochemistry and fluorescence microscopy suggests similar sensitivity but a more surface-confined identification due to the shallow penetration depth of the ToF-SIMS signal. The recorded ToF-SIMS images thus display the localization of lipids and amyloid-β in a narrow (~10 nm) two-dimensional plane at the tissue surface. As compared to a frozen nontreated tissue sample, the liposome preparation protocol generally increased the signal intensity of endogenous lipids, likely caused by matrix effects associated with the removal of salts, but no severe effects on the tissue integrity and the spatial distribution of lipids were observed with ToF-SIMS or scanning electron microscopy (SEM). This method may provide an important extension to conventional tissue imaging techniques to investigate the complex interplay of different kinds of molecules in neurodegenerative diseases, in the same specimen. However, limitations in target accessibility of the liposomes as well as unspecific binding need further consideration.


Immunotherapy and therapeutic vaccines in prostate cancer: an update on current strategies and clinical implications.
In recent years, immunotherapy has emerged as a viable and attractive strategy for the treatment of prostate cancer. While there are multiple ways to target the immune system, therapeutic cancer vaccines and immune checkpoint inhibitors have been most successful in late-stage clinical trials. The landmark Food and Drug Administration approval of sipuleucel-T for asymptomatic or minimally symptomatic metastatic prostate cancer set the stage for ongoing phase III trials with the cancer vaccine PSA-TRICOM and the immune checkpoint inhibitor ipilimumab. A common feature of these immune-based therapies is the appearance of improved overall survival without short-term changes in disease progression. This class effect appears to be due to modulation of tumor growth rate kinetics, in which the activated immune system exerts constant immunologic pressure that slows net tumor growth. Emerging data suggest that the ideal population for clinical trials of cancer vaccines is patients with lower tumor volume and less aggressive disease. Combination strategies that combine immunotherapy with standard therapies have been shown to augment both immune response and clinical benefit.


Metformin use in women with polycystic ovary syndrome.
Polycystic ovary syndrome (PCOS) is an endocrinopathy characterised by increased resistance to insulin. Metformin is one of the longest established oral insulin sensitising agents. For decades its use was restricted to management of type 2 diabetes. However, in the past two decades, its properties as an insulin sensitising agent have been explored in relation to its applicability for women with PCOS. Metformin is an effective ovulation induction agent for non-obese women with PCOS and offers some advantages over other first line treatments for anovulatory infertility such as clomiphene. For clomiphene-resistant women, metformin alone or in combination with clomiphene is an effective next step. Women with PCOS undergoing in vitro fertilisation should be offered metformin to reduce their risk of ovarian hyperstimulation syndrome. Limited evidence suggests that metformin may be a suitable alternative to the oral contraceptive pill (OCP) for treating hyperandrogenic symptoms of PCOS including hirsutism and acne. More research is required to define whether metformin has a role in improving long term health outcomes for women with PCOS, including the prevention of diabetes, cardiovascular disease and endometrial cancer.


Enhancing the Biodiversity of Ditches in Intensively Managed UK Farmland.
Drainage ditches, either seasonally flooded or permanent, are commonly found on intensively managed lowland farmland in the UK. They are potentially important for wetland biodiversity but, despite their ubiquity, information on their biodiversity and management in the wider countryside is scarce. We surveyed 175 ditches for their physical and chemical characteristics, spatial connectivity, plant communities and aquatic invertebrates in an area of intensively managed farmland in Oxfordshire, UK and collected information on ditch management from farmer interviews. Water depth and shade had a small impact on the diversity of plant and invertebrate communities in ditches. Increased shade over the ditch channel resulted in reduced taxonomic richness of both channel vegetation and aquatic invertebrates and channel vegetation cover was lower at shaded sites. Invertebrate taxonomic richness was higher when water was deeper. Spatial connectivity had no detectable impact on the aquatic invertebrate or plant communities found in ditches. The number of families within the orders Ephemeroptera, Plecoptera and Trichoptera (EPT), which contain many pollution-sensitive species, declined with decreasing pH of ditch water. As time since dredging increased, the number of EPT families increased in permanent ditches but decreased in temporary ditches. Whether or not a ditch was in an agri-environment scheme had little impact on the reported management regime or biodiversity value of the ditch. Measures for increasing the amount of water in ditches, by increasing the water depth or promoting retention of water in ditches, could increase the biodiversity value of ditches in agricultural land. Some temporary ditches for specialised species should be retained. Reducing the amount of shade over narrow ditches by managing adjacent hedgerows is also likely to increase the species diversity of plant and invertebrate communities within the ditch. We recommend that to preserve or enhance the biodiversity value of ditches, and improve their ecosystem service delivery, management prescriptions for hedgerows adjacent to ditches should differ from those aimed at hedgerows only.


Has the use of cervical, breast, and colorectal cancer screening increased in the United States?
This report describes trends in reported breast, cervical, and colorectal cancer screening within the US population from 1987 to 1992. Data from the 1987 and 1992 Cancer Control Supplements of the National Health Interview Survey were analyzed to determine use of Pap smears by women aged 18+; of mammography and clinical breast examination by women aged 50+; and of proctoscopy, digital rectal examination, and fecal occult blood testing among men and women aged 50+. Use of mammography doubled between 1987 and 1992 while Pap smear use changed very little. Use of the three colorectal cancer screening modalities increased but levels remained low. Usage trends were also assessed in relation to several sociodemographic factors. Disparities in screening reported in 1987 according to income and education persisted in 1992.


Genetic characteristics of non-familial epilepsy.
Knowledge of the genetic etiology of epilepsy can provide essential prognostic information and influence decisions regarding treatment and management, leading us into the era of precision medicine. However, the genetic basis underlying epileptogenesis or epilepsy pharmacoresistance is not well-understood, particularly in non-familial epilepsies with heterogeneous phenotypes that last until or start in adulthood. We sought to determine the contribution of known epilepsy-associated genes (EAGs) to the causation of non-familial epilepsies with heterogeneous phenotypes and to the genetic basis underlying epilepsy pharmacoresistance. We performed a multi-center study for whole exome sequencing-based screening of 178 selected EAGs in 243 non-familial adult patients with primarily focal epilepsy (122 drug-resistant and 121 drug-responsive epilepsies). The pathogenicity of each variant was assessed through a customized stringent filtering process and classified according to the American College of Medical Genetics and Genomics guidelines. Possible causal genetic variants of epilepsy were uncovered in 13.2% of non-familial patients with primarily focal epilepsy. The diagnostic yield according to the seizure onset age was 25% (2/8) in the neonatal and infantile period, 11.1% (14/126) in childhood and 14.7% (16/109) in adulthood. The higher diagnostic yields were from ion channel-related genes and mTOR pathway-related genes, which does not significantly differ from the results of previous studies on familial or early-onset epilepsies. These potentially pathogenic variants, which were identified in genes that have been mainly associated with early-onset epilepsies with severe phenotypes, were also linked to epilepsies that start in or last until adulthood in this study. This finding suggested the presence of one or more disease-modifying factors that regulate the onset time or severity of epileptogenesis. The target hypothesis of epilepsy pharmacoresistance was not verified in our study. Instead, neurodevelopment-associated epilepsy genes, such as TSC2 or RELN, or structural brain lesions were more strongly associated with epilepsy pharmacoresistance. We revealed a fraction of possible causal genetic variants of non-familial epilepsies in which genetic testing is usually overlooked. In this study, we highlight the importance of earlier identification of the genetic etiology of non-familial epilepsies, which leads us to the best treatment options in terms of precision medicine and to future neurobiological research for novel drug development. This should be considered a justification for physicians determining the hidden genetics of non-familial epilepsies that last until or start in adulthood.


Gender differences in diversion among non-medical users of prescription opioids and sedatives.
Non-medical use of prescription drugs is a major public health concern in the United States. Prescription opioids and sedatives are among the most widely abused drugs and their combined use can be lethal. Increasingly rigid prescribing guidelines may contribute to the changing context of opioid use and increase drug diversion. To examine gender differences in diversion of prescription opioids and sedatives among non-medical prescription opioid and sedative polysubstance users. We hypothesize that men will be more likely than women to engage in incoming diversion. Data from the Prescription Drug Abuse, Misuse, and Dependence Study, a cross-sectional study focused on prescription drug users, were analyzed. Non-medical use was defined as use of a drug that was not prescribed or use in a way other than prescribed. Individuals who reported past 12-month non-medical opioid and sedative use were included; diversion was defined as incoming (obtaining drugs from a source other than a health professional) and outgoing (giving away/selling/trading prescription drugs). Among the 198 polysubstance users, 41.4% were female. Men were 2.85 times as likely as women to report incoming diversion (95% CI: 1.21-6.72). Women were more likely to obtain opioids from a healthcare professional; men were more likely to obtain sedatives from a roommate, coworker, or friend. Over half of men and women reported outgoing diversion opioids or sedatives. Drug diversion highlights an important point of intervention. Current prevention efforts that target prescribers should be expanded to include users and diversion activities; these interventions should be gender-specific.


Evaluating comparative and equality judgments in contrast perception: attention alters appearance.
Covert attention not only improves performance in many visual tasks but also modulates the appearance of several visual features. Studies on attention and appearance have assessed subjective appearance using a task contingent upon a comparative judgment (e.g., M. Carrasco, S. Ling, & S. Read, 2004). Recently, K. A. Schneider and M. Komlos (2008) questioned the validity of those results because they did not find a significant effect of attention on contrast appearance using an equality task. They claim that such equality judgments are bias-free whereas comparative judgments are bias-prone and propose an alternative interpretation of the previous findings based on a decision bias. However, to date there is no empirical support for the superiority of the equality procedure. Here, we compare biases and sensitivity to shifts in perceived contrast of both paradigms. We measured contrast appearance using both a comparative and an equality judgment. Observers judged the contrasts of two simultaneously presented stimuli, while either the contrast of one stimulus was physically incremented (Experiments 1 and 2) or exogenous attention was drawn to it (Experiments 3 and 4). We demonstrate several methodological limitations of the equality paradigm. Nevertheless, both paradigms capture shifts in PSE due to physical and perceived changes in contrast and show that attention enhances apparent contrast.


Therapeutic potential of rat monoclonal antibodies: isotype specificity of antibody-dependent cell-mediated cytotoxicity with human lymphocytes.
The ability of rat monoclonal antibodies to promote antibody-dependent cell-mediated cytotoxicity with human effector cells was tested by using a variety of antibodies against different human and mouse leukocyte antigens. It was found that only IgG2b antibodies were effective. This isotype has already been shown to be efficient in fixing human complement, which suggests that among rat monoclonal antibodies, the IgG2b subclass might be a good choice for attempts at serotherapy. Further studies with other antibody-mediated effector mechanisms as well as suitable clinical trials are merited.


Comparison of two exploratory data analysis methods for classification of Phyllanthus chemical fingerprint: unsupervised vs. supervised pattern recognition technologies.
In this study, unsupervised and supervised classification methods were compared for comprehensive analysis of the fingerprints of 26 Phyllanthus samples from different geographical regions and species. A total of 63 compounds were identified and tentatively assigned structures for the establishment of fingerprints using high-performance liquid chromatography time-of-flight mass spectrometry (HPLC/TOFMS). Unsupervised and supervised pattern recognition technologies including principal component analysis (PCA), nearest neighbors algorithm (NN), partial least squares discriminant analysis (PLS-DA), and artificial neural network (ANN) were employed. Results showed that Phyllanthus could be correctly classified according to their geographical locations and species through ANN and PLS-DA. Important variables for clusters discrimination were also identified by PCA. Although unsupervised and supervised pattern recognitions have their own disadvantage and application scope, they are effective and reliable for studying fingerprints of traditional Chinese medicines (TCM). These two technologies are complementary and can be superimposed. Our study is the first holistic comparison of supervised and unsupervised pattern recognition technologies in the TCM chemical fingerprinting. They showed advantages in sample classification and data mining, respectively.


Percolating ideas: The effects of caffeine on creative thinking and problem solving.
Caffeine is the most widely consumed psychotropic drug in the world, with numerous studies documenting the effects of caffeine on people's alertness, vigilance, mood, concentration, and attentional focus. The effects of caffeine on creative thinking, however, remain unknown. In a randomized placebo-controlled between-subject double-blind design the present study investigated the effect of moderate caffeine consumption on creative problem solving (i.e., convergent thinking) and creative idea generation (i.e., divergent thinking). We found that participants who consumed 200 mg of caffeine (approximately one 12 oz cup of coffee, n = 44), compared to those in the placebo condition (n = 44), showed significantly enhanced problem-solving abilities. Caffeine had no significant effects on creative generation or on working memory. The effects remained after controlling for participants' caffeine expectancies, whether they believed they consumed caffeine or a placebo, and changes in mood. Possible mechanisms and future directions are discussed.


Surgical Updates in the Treatment of Pelvic Organ Prolapse.
Pelvic organ prolapse affects approximately 8% of women, and the demand for pelvic organ prolapse surgery is expected to increase by nearly 50% over the next 40 years. The surgical techniques used to correct pelvic organ prolapse have evolved over the last 10 years, with multiple well-designed studies addressing the risks, outcomes, reoperation rates, and optimal surgical approaches. Here we review the most recent evidence on the route of access, concomitant procedures, and synthetic materials for augmenting the repair. Ultimately, this review highlights that there is no optimal method for correcting pelvic organ prolapse and that the risks, benefits, and approaches should be discussed in a patient-centered, goal-oriented approach to decision-making.


Microvessel Chaste: An Open Library for Spatial Modeling of Vascularized Tissues.
Spatial models of vascularized tissues are widely used in computational physiology. We introduce a software library for composing multiscale, multiphysics models for applications including tumor growth, angiogenesis, osteogenesis, coronary perfusion, and oxygen delivery. Composition of such models is time consuming, with many researchers writing custom software. Recent advances in imaging have produced detailed three-dimensional (3D) datasets of vascularized tissues at the scale of individual cells. To fully exploit such data there is an increasing need for software that allows user-friendly composition of efficient, 3D models of vascularized tissues, and comparison of predictions with in vivo or in vitro experiments and alternative computational formulations. Microvessel Chaste can be used to build simulations of vessel growth and adaptation in response to mechanical and chemical stimuli; intra- and extravascular transport of nutrients, growth factors and drugs; and cell proliferation in complex 3D geometries. In addition, it can be used to develop custom software for integrating modeling with experimental data processing workflows, facilitated by a comprehensive Python interface to solvers implemented in C++. This article links to two reproducible example problems, showing how the library can be used to build simulations of tumor growth and angiogenesis with realistic vessel networks.


Clinical and microbiological findings in patients with peri-implantitis: a cross-sectional study.
The aim of this study was to analyze clinical and microbiological characters in subjects and implants affected and not affected by peri-implantitis. Additionally, same features were analyzed also intra-individually, comparing healthy and diseased implants within the same subject. A total of 534 patients who received at least 1 implant and coming to routine check-up or spontaneous visits at the University of Valencia were recruited. Clinical parameters including Bleeding on probing (BoP), Probing pocket depth (PPD), and Pi were screened. Samples for microbiological analysis were obtained from three locations: peri-implant sulci (PIS), inner parts of the implant connections (I), and gingival sulci of neighboring teeth (GS). Quantitative real-time PCR was performed for total counts of 10 microorganisms. A total of 534 patients with 1507 dental implants were analyzed. The prevalence of peri-implantitis was found 10.3% for patients and 7.3% for implants. Higher percentage of healthy periodontal subjects were found in the non-peri-implantitis group. The analysis within the 53 patients affected by peri-implantitis revealed that the implants affected by peri-implantitis presented a higher percentage of plaque, BoP, and number of implants presenting <2 mm attached gingiva. Additionally, more cemented crowns and implants inserted in bone-augmented sites were found among the diseased implants. The microbiologic analysis presented no relevant differences between the analysis at the peri-implant sulcus (PIS) and the connections inside the abutments surfaces (PI). The microbial composition at the neighboring teeth (GS) resembled the composition found at the PIS with a high frequency of Pg, Tf, Pi, PM, and Ec. The results of this study seem to indicate that inadequate oral hygiene and the presence of bleeding from the gingiva/mucosa in patients with dental implant were associated with an higher prevalence of peri-implantitis; moreover, in the patients affected by peri-implantitis, the lack of sufficient height keratinized mucosa (<2 mm) and bone regenerative procedures at implant level were also associated to higher prevalence of peri-implantitis as well.


An SCL 3' enhancer targets developing endothelium together with embryonic and adult haematopoietic progenitors.
The SCL gene encodes a basic helix-loop-helix transcription factor which is expressed in early haematopoietic progenitors throughout ontogeny and is essential for the normal development of blood and blood vessels. Transgenic studies have characterised spatially distinct 5' enhancers which direct lacZ expression to subdomains of the normal SCL expression pattern, but the same elements failed to produce appropriate haematopoietic expression. We now describe an SCL 3' enhancer with unique properties. It directed lacZ expression in transgenic mice to extra-embryonic mesoderm and subsequently to both endothelial cells and to a subset of blood cells at multiple sites of embryonic haematopoiesis including the yolk sac, para-aortic splanchnopleura and AGM region. The 3' enhancer also targeted expression to haematopoietic progenitors in both foetal liver and adult bone marrow. Purified lacZ(+ )cells were highly enriched for clonogenic myeloid and erythroid progenitors as well as day-12 spleen colony forming units (CFU-S). Within the total gated population from bone marrow, 95% of the myeloid and 90% of the erythroid colony-forming cells were contained in the lacZ(+) fraction, as were 98% of the CFU-S. Activation of the enhancer did not require SCL protein. On the contrary, transgene expression in yolk sacs was markedly increased in an SCL-/- background, suggesting that SCL is subject to negative autoregulation. Alternatively the SCL-/- environment may alter differentiation of extra-embryonic mesoderm and result in an increased number of cells capable of expressing high levels of the transgene. Our data represents the first description of an enhancer that integrates information necessary for expression in developing endothelium and early haematopoietic progenitors at distinct times and sites throughout ontogeny. This enhancer provides a potent tool for the manipulation of haematopoiesis and vasculogenesis in vivo.


Optimisation of a 2-D gel electrophoresis protocol for the human-pathogenic fungus Aspergillus fumigatus.
Aspergillus fumigatus is the most important airborne fungal pathogen causing life-threatening infections in immunosuppressed patients. One of the important questions concerning A. fumigatus is the identification of pathogenicity determinants. To obtain a comprehensive overview about the proteins produced at different physiological conditions that are related to the infectious process a proteomic approach has been applied. Here, 2-D gel electrophoresis for filamentous fungi was optimised concerning removal of interfering compounds, protein extraction and separation methods. A trichloroacetic acid-based precipitation method of proteins with their subsequent solubilisation by the use of a combination of CHAPS with a second sulfobetaine detergent gave the best results. The optimised protocol was evaluated by the analysis of the proteomes of A. fumigatus grown on two different carbon sources, i.e., glucose and ethanol. Carbon catabolite repression has not been studied in detail at the protein level in A. fumigatus yet. In addition, growth on ethanol leads to activation of the glyoxylate cycle which was shown to be essential for pathogenesis in bacteria and fungi. In A. fumigatus, differential patterns of enzymes of the gluconeogenesis, glyoxylate cycle and ethanol degradation pathway during growth on glucose and ethanol were observed.


Rod sensitivity, cone sensitivity, and photoreceptor layer thickness in retinal degenerative diseases.
To evaluate the effects of selective rod and/or cone loss on frequency-domain optical coherence tomography (fdOCT) measures of photoreceptor structure in patients with retinal degenerative diseases. Six patients with cone dystrophy (CD) and eight patients with retinitis pigmentosa (RP) were recruited from the Southwest Eye Registry on the basis of diagnosis and ERG findings. fdOCT horizontal line scans were segmented to obtain the thicknesses of the outer segments plus RPE (OS+) and the outer nuclear layer (ONL). The normalized product ONL*OS was obtained after dividing by mean ONL*OS values of 23 normal individuals. Visual field sensitivity profiles were obtained with a modified retinal perimeter, from the horizontal midline with short- and long-wave stimuli under dark- and light-adapted conditions. Patients with CD and normal rod-mediated sensitivity, but decreased cone-mediated sensitivity, showed normal ONL*OS outside the fovea. The total receptor layer was thinned in the fovea, consistent with loss in cone nuclei and Henle's fiber layer. Patients with RP and sensitivity in the dark that was mediated by cones showed ONL*OS thickness that was linearly related to cone sensitivity. ONL*OS thickness was linearly related to rod sensitivity in regions with greater loss of cone than rod sensitivity. Both rods and cones can support an intact IS/OS junction and normal photoreceptor thickness measures. The product of ONL and OS thicknesses is proportional to the sensitivity mediated by the less abnormal type of photoreceptor.


All-optical switching of nematic liquid crystal films driven by localized surface plasmons.
We have demonstrated an all-optical technique for reversible in-plane and out-of-plane switching of nematic liquid crystal molecules in few micron thick films. Our method leverages the highly localized electric fields ("hot spots") and plasmonic heating that are generated in the near-field region of densely packed gold nanoparticle layers optically excited on-resonance with the localized surface plasmon absorption. Using polarized microscopy and transmission measurements, we observe this switching from homeotropic to planar over a temperature range starting at room temperature to just below the isotropic transition, and at on-resonance excitation intensity less than 0.03 W/cm(2). In addition, we controllably vary the in-plane directionality of the liquid crystal molecules in the planar state by altering the linear polarization of the incident excitation. Using discrete dipole simulations and control measurements, we establish spectral selectivity in this new and interesting perspective for photonic application using low light power.


Trauma-informed care for children in the ambulance: international survey among pre-hospital providers.
Background: Pre-hospital providers, such as paramedics and emergency medical technicians, are in a position to provide key emotional support to injured children and their families. Objective: Our goal was to examine (a) pre-hospital providers' knowledge of traumatic stress in children, attitudes towards psychosocial aspects of care, and confidence in providing psychosocial care, (b) variations in knowledge, attitudes, and confidence according to demographic and professional characteristics, and (c) training preferences of pre-hospital providers regarding psychosocial care to support paediatric patients and their families. Method: We conducted a cross-sectional, online survey among an international sample of 812 pre-hospital providers from high-income countries. The questionnaire was adapted from a measure for a similar study among Emergency Department staff, and involved 62 items in 7 main categories (e.g. personal and work characteristics, knowledge of paediatric traumatic stress, and confidence regarding 18 elements of psychosocial care). The main analyses comprised descriptive statistics and multiple regression analyses. Results: On average, respondents answered 2.7 (SD = 1.59) out of seven knowledge questions correctly. Respondents with higher knowledge scores were more often female, parent of a child under 17, and reported that at least 10% of their patients were children. A majority of participants (83.5%) saw all 18 aspects of psychosocial care as part of their job. Providers felt moderately confident (M = 3.2, SD = 0.45) regarding their skills in psychosocial care, which was predicted by gender (female), having more experience, having a larger proportion of child patients, and having received training in psychosocial care in the past five years. Most respondents (89.7%) wanted to gain more knowledge and skills regarding psychosocial care for injured children. In terms of training format, they preferred an interactive website or a one-off group training. Conclusions: There appears to be both a need and an opportunity for education initiatives regarding paediatric traumatic stress in the pre-hospital context.


The Experiences of Specialist Nurses Working Within the Uro-oncology Multidisciplinary Team in the United Kingdom.
United Kingdom prostate cancer nursing care is provided by a variety of urology and uro-oncology nurses. The experience of working in multidisciplinary teams (MDT) was investigated in a national study. The study consisted of a national survey with descriptive statistics and thematic analysis. A secondary analysis of a data subset from a UK whole population survey was undertaken (n = 285) of the specialist nursing workforce and the services they provide. Data were collected on the experience of working in the MDT. Forty-five percent of the respondents felt that they worked in a functional MDT, 12% felt that they worked in a dysfunctional MDT, and 3.5% found the MDT meeting intimidating. Furthermore, 34% of the nurses felt that they could constructively challenge all members of the MDT in meetings. Themes emerging from open-ended questions were lack of interest in nonmedical concerns by other team members, ability to constructively challenge decisions or views within the meeting, and little opportunity for patients' wishes to be expressed. Despite expertise and experience, nurses had a variable, often negative, experience of the MDT. It is necessary to ensure that all participants can contribute and are heard and valued. More emphasis should be given to patients' nonmedical needs.


The tacrolimus metabolism rate influences renal function after kidney transplantation.
The effective calcineurin inhibitor (CNI) tacrolimus (Tac) is an integral part of the standard immunosuppressive regimen after renal transplantation (RTx). However, as a potent CNI it has nephrotoxic potential leading to impaired renal function in some cases. Therefore, it is of high clinical impact to identify factors which can predict who is endangered to develop CNI toxicity. We hypothesized that the Tac metabolism rate expressed as the blood concentration normalized by the dose (C/D ratio) is such a simple predictor. Therefore, we analyzed the impact of the C/D ratio on kidney function after RTx. Renal function was analyzed 1, 2, 3, 6, 12 and 24 months after RTx in 248 patients with an immunosuppressive regimen including basiliximab, tacrolimus, mycophenolate mofetil and prednisolone. According to keep the approach simple, patients were split into three C/D groups: fast, intermediate and slow metabolizers. Notably, compared with slow metabolizers fast metabolizers of Tac showed significantly lower estimated glomerular filtration rate (eGFR) values at all the time points analyzed. Moreover, fast metabolizers underwent more indication renal biopsies (p = 0.006) which revealed a higher incidence of CNI nephrotoxicity (p = 0.015) and BK nephropathy (p = 0.024) in this group. We herein identified the C/D ratio as an easy calculable risk factor for the development of CNI nephrotoxicity and BK nephropathy after RTx. We propose that the simple C/D ratio should be taken into account early in patient's risk management strategies.


Next generation sequencing of CLU, PICALM and CR1: pitfalls and potential solutions.
CLU, PICALM and CR1 were identified as genetic risk factors for late onset Alzheimer's disease (AD) in two large genome wide association studies (GWAS) published in 2009, but the variants that convey this alteration in disease risk, and how the genes relate to AD pathology is yet to be discovered. A next generation sequencing (NGS) project was conducted targeting CLU, CR1 and PICALM, in 96 AD samples (8 pools of 12), in an attempt to discover rare variants within these AD associated genes. Inclusion of repetitive regions in the design of the SureSelect capture lead to significant issues in alignment of the data, leading to poor specificity and a lower than expected depth of coverage. A strong positive correlation (0.964, p<0.001) was seen between NGS and 1000 genome project frequency estimates. Of the ~170 "novel" variants detected in the genes, seven SNPs, all of which were present in multiple sample pools, were selected for validation by Sanger sequencing. Two SNPs were successfully validated by this method, and shown to be genuine variants, while five failed validation. These spurious SNP calls occurred as a result of the presence of small indels and mononucleotide repeats, indicating such features should be regarded with caution, and validation via an independent method is important for NGS variant calls.


Osteonecrosis of the Jaws in Dogs in Previously Irradiated Fields: 13 Cases (1989-2014).
The aim of this report was to characterize osteonecrosis of the jaws (ONJ) in previously irradiated fields in dogs that underwent radiotherapy (RT) for oral tumors. Osteoradionecrosis of the jaw (ORNJ) was further defined as osteonecrosis in a previously irradiated field in the absence of a tumor. Thirteen dogs clinically diagnosed with 15 ONJ lesions were included in this retrospective case series. Medical records were reviewed for: breed, sex, weight, and age of the patient, tumor type, location in the oral cavity and size, location of the ONJ, time from RT to ONJ onset, known duration of the ONJ, and tumor presence. Where available, histological assessment of tissues obtained from the primary tumor, and tissues obtained from the ONJ lesion, was performed, and computed tomographic (CT) images and dental radiographs were reviewed. RT and other treatment details were also reviewed. Twelve dogs developed ONJ in the area of the previously irradiated tumor or the jaw closest to the irradiated mucosal tumor. Recurrence of neoplasia was evident at the time of ONJ diagnosis in five dogs. Time from RT start to ONJ onset varied from 2 to 44 months. In three cases, ORNJ developed after dental extractions in the irradiated field. Dental radiographs mostly revealed a moth-eaten pattern of bone loss, CT mostly revealed osteolysis, and histopathology was consistent with osteonecrosis. To conclude, development of ONJ/ORNJ following RT is a rare, but potentially fatal complication. Patients undergoing RT may benefit from a comprehensive oral and dental examination and treatment prior to RT.


Mechanistic Modeling of Genetic Circuits for ArsR Arsenic Regulation.
Bioreporters are living cells that generate an easily measurable signal in the presence of a chemical compound. They acquire their functionality from synthetic gene circuits, the configuration of which defines the response signal and signal-to-noise ratio. Bioreporters based on the Escherichia coli ArsR system have raised significant interest for quantifying arsenic pollution, but they need to be carefully optimized to accurately work in the required low concentration range (1-10 μg arsenite L-1). To better understand the general functioning of ArsR-based genetic circuits, we developed a comprehensive mechanistic model that was empirically tested and validated in E. coli carrying different circuit configurations. The model accounts for the different elements in the circuits (proteins, DNA, chemical species), and their detailed affinities and interactions, and predicts the (fluorescent) output from the bioreporter cell as a function of arsenite concentration. The model was parametrized using existing ArsR biochemical data, and then complemented by parameter estimations from the accompanying experimental data using a scatter search algorithm. Model predictions and experimental data were largely coherent for feedback and uncoupled circuit configurations, different ArsR alleles, promoter strengths, and presence or absence of arsenic efflux in the bioreporters. Interestingly, the model predicted a particular useful circuit variant having steeper response at low arsenite concentrations, which was experimentally confirmed and may be useful as arsenic bioreporter in the field. From the extensive validation we expect the mechanistic model to further be a useful framework for detailed modeling of other synthetic circuits.


Hepatic Transcriptome Analysis Identifies Divergent Pathogen-Specific Targeting-Strategies to Modulate the Innate Immune System in Response to Intramammary Infection.
Mastitis is one of the major risks for public health and animal welfare in the dairy industry. Two of the most important pathogens to cause mastitis in dairy cattle are Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). While S. aureus generally induces a chronic and subclinical mastitis, E. coli is an important etiological pathogen resulting in an acute and clinical mastitis. The liver plays a central role in both, the metabolic and inflammatory physiology of the dairy cow, which is particularly challenged in the early lactation due to high metabolic and immunological demands. In the current study, we challenged the mammary glands of Holstein cows with S. aureus or E. coli, respectively, mimicking an early lactation infection. We compared the animals' liver transcriptomes with those of untreated controls to investigate the hepatic response of the individuals. Both, S. aureus and E. coli elicited systemic effects on the host after intramammary challenge and seemed to use pathogen-specific targeting strategies to bypass the innate immune system. The most striking result of our study is that we demonstrate for the first time that S. aureus intramammary challenge causes an immune response beyond the original local site of the mastitis. We found that in the peripheral liver tissue defined biological pathways are switched on in a coordinated manner to balance the immune response in the entire organism. TGFB1 signaling plays a crucial role in this context. Important pathways involving actin and integrin, key components of the cytoskeleton, were downregulated in the liver of S. aureus infected cows. In the hepatic transcriptome of E. coli infected cows, important components of the complement system were significantly lower expressed compared to the control cows. Notably, while S. aureus inhibits the cell signaling by Rho GTPases in the liver, E. coli switches the complement system off. Also, metabolic hepatic pathways (e.g., lipid metabolism) are affected after mammary gland challenge, demonstrating that the liver restricts metabolic tasks in favor of the predominant immune response after infection. Our results provide new insights for the infection-induced modifications of the dairy cow's hepatic transcriptome following mastitis.


Triiodothyronine enhances renal response to aldosterone in the rabbit collecting tubule.
Since thyroid hormones and mineralocorticoids were observed to stimulate kidney Na-K-ATPase in similar sites and with similar time courses, this study was initiated to evaluate whether aldosterone is involved in the stimulation of Na-K-ATPase observed in collecting tubules 3 h after triiodothyronine (T3) administration to thyroidectomized (TX) rabbits. Results indicate that: Plasma aldosterone level decreased markedly in TX rabbits but was not restored 3 h after T3 injection; Early stimulation of Na-K-ATPase by T3 was abolished when plasma aldosterone level was suppressed by adrenalectomy or when aldosterone effects were blocked by spironolactone; Administration of aldosterone to TX rabbits mimicked the action of T3; Sensitivity of Na-K-ATPase to aldosterone markedly decreased after thyroidectomy. These results demonstrate an interaction between aldosterone and T3 in the control of Na-K-ATPase in the collecting tubule. Triiodothyronine enhances the sensitivity of Na-K-ATPase to aldosterone which, in turn, produces a stimulatory action despite the decreased plasma level observed during hypothyroidism.


High dose multiple micronutrient supplementation improves villous morphology in environmental enteropathy without HIV enteropathy: results from a double-blind randomised placebo controlled trial in Zambian adults.
Environmental enteropathy (EE) is an asymptomatic abnormality of small bowel structure and function, which may underlie vaccine inefficacy in the developing world. HIV infection co-exists in many of these populations. There is currently no effective treatment. We conducted a secondary analysis of a randomised controlled trial of high dose multiple micronutrient (MM) supplementation on small bowel architecture in EE in participants with or without HIV infection. In a double-blind parallel-group trial of the effect of MM on innate immune responses to oral vaccines, consenting Zambian adults were randomised to receive 6 weeks of 24 micronutrients as a daily capsule or placebo. HIV status was established after randomisation. Proximal jejunal biopsies were obtained after the supplementation period. Villous height, crypt depth, villous width, villous perimeter per 100 μm muscularis mucosa (a measure of epithelial surface area), and villous cross sectional area per 100 μm muscularis mucosa (a measure of villous compartment volume) were measured in orientated biopsy sections using semi-automated image analysis. Analysis was by intention to treat. 18 patients received MM and 20 placebo. 6/18 MM and 9/20 placebo patients had HIV. In HIV negative patients given MM compared to placebo, mean villous height was 24.0% greater (293.3 v. 236.6 μm; 95% CI of difference 17.7-95.9 μm; P = 0.006), mean villous area was 27.6% greater (27623 v. 21650 μm2/100 μm; 95% CI of difference 818-11130 μm2/100 μm; P = 0.03), and median villous perimeter was 29.7% greater (355.0 v. 273.7 μm/100 μm; 95% CI of difference 16.3-146.2 μm/100 μm; P = 0.003). There was no significant effect on crypt depth or villous width. No effect was observed in HIV positive patients. There were no adverse events attributable to MM. MM improved small bowel villous height and absorptive area, but not crypt depth, in adults with EE without HIV. Nutritional intervention may therefore selectively influence villous compartment remodelling. In this small study, there was a clear difference in response depending on HIV status, suggesting that EE with superimposed HIV enteropathy may be a distinct pathophysiological condition.


Understanding Between-Person Interventions With Time-Intensive Longitudinal Outcome Data: Longitudinal Mediation Analyses.
Mediation analysis is an important tool for understanding the processes through which interventions affect health outcomes over time. Typically the temporal intervals between X, M, and Y are fixed by design, and little focus is given to the temporal dynamics of the processes. In this article, we aim to highlight the importance of considering the timing of the causal effects of a between-person intervention X, on M and Y, resulting in a deeper understanding of mediation. We provide a framework for examining the impact of a between-person intervention X on M and Y over time when M and Y are measured repeatedly. Five conceptual and analytic steps involve visualizing the effects of the intervention on Y, M, the relationship of M and Y, and the mediating process over time and selecting an appropriate analytic model. We demonstrate how these steps can be applied to two empirical examples of health behavior change interventions. We show that the patterns of longitudinal mediation can be fit with versions of longitudinal multilevel structural equation models that represent how the magnitude of direct and indirect effects vary over time. We urge researchers and methodologists to pay more attention to temporal dynamics in the causal analysis of interventions.


A case-control study of risk factors for Haemophilus influenzae type B disease in Navajo children.
To understand the potential risk factors and protective factors for invasive Haemophilus influenzae type b (Hib) disease, we conducted a case-control study among Navajo children less than two years of age resident on the Navajo Nation. We analyzed household interview data for 60 cases that occurred between August 1988 and February 1991, and for 116 controls matched by age, gender, and geographic location. The Hib vaccine recipients were excluded from the analyses. Conditional logistic regression models were fit to examine many variables relating to social and environmental conditions. Risk factors determined to be important were never breast fed (odds ratio [OR] = 3.55, 95% confidence interval [CI] = 1.52, 8.26), shared care with more than one child less than two years of age (OR = 2.32, 95% CI = 0.91, 5.96); wood heating (OR = 2.14, 95% CI = 0.91, 5.05); rodents in the home (OR = 8.18, 95% CI = 0.83, 80.7); and any livestock near the home (OR = 2.18, 95% CI = 0.94, 5.04).


Long-Term Immunogenicity and Safety of a Conventional Influenza Vaccine in Patients with Type 2 Diabetes.
No previous studies have assessed the persistence of immune responses in individuals with diabetes. We conducted this study to evaluate the long-term immunogenicity and safety of an influenza vaccine in type 2 diabetic subjects compared with nondiabetic controls. A randomized and controlled study was conducted at two university hospitals during the 2012-2013 influenza season. The study vaccine was a standard-dose trivalent subunit inactivated intramuscular vaccine. Serum hemagglutination-inhibiting (HI) antibodies were measured at the time of vaccination and 1 month and 6 months after vaccination. Local and systemic reactions were recorded for 7 days. A total of 105 diabetic patients and 108 controls were included in the analysis. One month after vaccination, both the diabetic and nondiabetic groups satisfied all of the criteria of the Committee for Medical Products for Human Use (CHMP), and the immunogenicity profiles were statistically similar between the two groups. Although the vaccine was well tolerated, and all adverse reactions were mild to moderate, there was a tendency toward a reduced incidence of local reactions in the diabetic group. All values in the long-term immunogenicity profiles were statistically similar between the two groups, except for the seroprotection rate for the A/H1N1 influenza virus strain, which was significantly lower in the elderly diabetic group than that in the elderly nondiabetic group. However, in multivariate analysis, long-term immunogenicity was associated with age and prevaccination titer, regardless of diabetes status. (This study has been registered at CRIS [https://cris.nih.go.kr/cris/en/] under registration no. KCT0001423.).


Trigger point acupuncture treatment of chronic low back pain in elderly patients--a blinded RCT.
There is some evidence for the efficacy of acupuncture in chronic low back pain, but it remains unclear which acupuncture modes are most effective. Our objective was to evaluate the effects of two different modes of trigger point acupuncture on pain and quality of life in chronic low back pain patients compared to standard acupuncture treatment. Thirty five consecutive out-patients (25 women, 10 men; age range: 65-81 years) from the Department of Orthopaedic Surgery, Meiji University of Oriental Medicine, with non-radiating low back pain for at least six months and normal neurological examination, were randomised to one of three groups over 12 weeks. Each group received two phases of acupuncture treatment with an interval between them. Nine patients dropped out during the course of the study. The standard acupuncture group (n=9) received treatment at traditional acupuncture points for low back pain, while the other acupuncture groups received superficial (n=9) or deep (n=9) treatments on trigger points. Outcome measures were VAS pain intensity and Roland Morris Questionnaire. After treatment, the group that received deep needling to trigger points reported less pain intensity and improved quality of life compared to the standard acupuncture group or the group that received superficial needling to trigger points, but the differences were not statistically significant. There was a significant reduction in pain intensity between the treatment and interval in the group that received deep needling to trigger points (P<0.01), but not in the standard acupuncture group or the group that received superficial needling to trigger points. These results suggest that deep needling to trigger points may be more effective in the treatment of low back pain in elderly patients than either standard acupuncture therapy, or superficial needling to trigger points.


Phase mutual information as a similarity measure for registration.
This article describes a new method for non-rigid alignment of multimodal images. Multimodal image registration is most often accomplished by modelling, in some sense, an intensity mapping between the images. Here, the alternative strategy of modelling a relationship between local image phase is introduced. This method is intrinsically image feature based, and searches for relationships between feature appearances, rather than tissue class intensity. This enables registration of modalities for which image intensity is not a simple function of tissue class, for example ultrasound. It is also demonstrated that this method performs comparably to an intensity method even when the images are related by a simple intensity transform, but that the phase method is significantly more robust to image artifacts which corrupt the ideal intensity mapping.


Lower leg muscle structure and function are altered in long-distance runners with medial tibial stress syndrome: a case control study.
Medial tibial stress syndrome (MTSS) is a common lower leg injury experienced by runners. Although numerous risk factors are reported in the literature, many are non-modifiable and management of the injury remains difficult. Lower leg muscle structure and function are modifiable characteristics that influence tibial loading during foot-ground contact. Therefore, this study aimed to determine whether long-distance runners with MTSS displayed differences in in vivo lower leg muscle structure and function than matched asymptomatic runners. Lower leg structure was assessed using ultrasound and a measure of lower leg circumference to quantify muscle cross-sectional area, thickness and lean lower leg girth. Lower leg function was assessed using a hand-held dynamometer to quantify maximal voluntary isometric contraction strength and a single leg heel raise protocol was used to measure ankle plantar flexor endurance. Outcome variables were compared between the limbs of long-distance runners suffering MTSS (n = 20) and matched asymptomatic controls (n = 20). Means, standard deviations, 95 % confidence intervals, mean differences and Cohen's d values were calculated for each variable for the MTSS symptomatic and control limbs. MTSS symptomatic limbs displayed a significantly smaller flexor hallucis longus cross-sectional area, a smaller soleus thickness but a larger lateral gastrocnemius thickness than the control limbs. However, there was no statistical difference in lean lower leg girth. Compared to the matched control limbs, MTSS symptomatic limbs displayed deficits in maximal voluntary isometric contraction strength of the flexor hallucis longus, soleus, tibialis anterior and peroneal muscles, and reduced ankle plantar flexor endurance capacity. Differences in lower leg muscle structure and function likely render MTSS symptomatic individuals less able to withstand the negative tibial bending moment generated during midstance, potentially contributing to the development of MTSS. The clinical implications of these findings suggest that rehabilitation protocols for MTSS symptomatic individuals should aim to improve strength of the flexor hallucis longus, soleus, tibialis anterior and peroneal muscles along with ankle plantar flexor endurance. However, the cross-sectional study design prevents us determining whether between group differences were a cause or effect of MTSS. Therefore, future prospective studies are required to substantiate the study findings.


Amperometric magnetoimmunosensor for ErbB2 breast cancer biomarker determination in human serum, cell lysates and intact breast cancer cells.
A highly sensitive amperometric magnetoimmunosensor for the determination of ErbB2 protein, a well-known biomarker related to high-impact high-incidence diseases such as breast cancer, is described. A sandwich format involving the covalent immobilization of a specific capture antibody onto magnetic beads (MBs) and incubation of the modified MBs with a mixture solution of the antigen and a HRP-labeled detector antibody was used. The resulting modified MBs were captured on the surface of a disposable screen-printed carbon electrode (SPCE) and the amperometric responses at -0.20 V were measured. This ErbB2 magnetoimmunosensor exhibited a very low detection limit of 26 pg mL(-1) far below the established cut-off for this biomarker (15 ng mL(-1)) and was successfully applied to the quantitation of ErbB2 in human serum and cell lysates samples without any matrix effect. In addition, the developed assay allowed the assessment of ErbB2 status directly in intact breast cancer cells. The results correlated well with those obtained with a commercial ELISA method, thus demonstrating that the new magnetoimmunosensing platform offers a truthful and useful analytical tool to be easily applied in breast cancer diagnosis through either ErbB2 protein determination or breast cancer cell status detection.


The first endogenous herpesvirus, identified in the tarsier genome, and novel sequences from primate rhadinoviruses and lymphocryptoviruses.
Herpesviridae is a diverse family of large and complex pathogens whose genomes are extremely difficult to sequence. This is particularly true for clinical samples, and if the virus, host, or both genomes are being sequenced for the first time. Although herpesviruses are known to occasionally integrate in host genomes, and can also be inherited in a Mendelian fashion, they are notably absent from the genomic fossil record comprised of endogenous viral elements (EVEs). Here, we combine paleovirological and metagenomic approaches to both explore the constituent viral diversity of mammalian genomes and search for endogenous herpesviruses. We describe the first endogenous herpesvirus from the genome of the Philippine tarsier, belonging to the Roseolovirus genus, and characterize its highly defective genome that is integrated and flanked by unambiguous host DNA. From a draft assembly of the aye-aye genome, we use bioinformatic tools to reveal over 100,000 bp of a novel rhadinovirus that is the first lemur gammaherpesvirus, closely related to Kaposi's sarcoma-associated virus. We also identify 58 genes of Pan paniscus lymphocryptovirus 1, the bonobo equivalent of human Epstein-Barr virus. For each of the viruses, we postulate gene function via comparative analysis to known viral relatives. Most notably, the evidence from gene content and phylogenetics suggests that the aye-aye sequences represent the most basal known rhadinovirus, and indicates that tumorigenic herpesviruses have been infecting primates since their emergence in the late Cretaceous. Overall, these data show that a genomic fossil record of herpesviruses exists despite their extremely large genomes, and expands the known diversity of Herpesviridae, which will aid the characterization of pathogenesis. Our analytical approach illustrates the benefit of intersecting evolutionary approaches with metagenomics, genetics and paleovirology.


In vivo induction of cytotoxic T cell response by a free synthetic peptide requires CD4+ T cell help.
Most attempts to induce CTL responses by in vivo priming with free synthetic peptides have been unsuccessful so far. However, two separate studies have recently succeeded in inducing antiviral CTL responses by immunizing mice with unmodified free synthetic peptides derived from nucleoproteins from either lymphocytic choriomeningitis virus or Sendai virus. In the present study, we have analyzed the cellular mechanisms by which the lymphocytic choriomeningitis virus synthetic peptide induced CTL responses. We demonstrated that this peptide, which was previously shown to be recognized by CD8+ T cells, also contains a helper CD4+ T cell epitope. It stimulates in vivo both CD4+ T cell-mediated CTL response. The in vivo elimination of CD4+ T cells by treatment with a mAb was shown to strongly reduce the antipeptide CTL response. This study therefore demonstrates that to be able to induce CTL responses, a peptide has to stimulate both CD4+ and CD8+ T cell subset.


Opine-based Agrobacterium competitiveness: dual expression control of the agrocinopine catabolism (acc) operon by agrocinopines and phosphate levels.
Agrobacterium tumefaciens strain C58 can transform plant cells to produce and secrete the sugar-phosphate conjugate opines agrocinopines A and B. The bacterium then moves in response to the opines and utilizes them as exclusive sources of carbon, energy, and phosphate via the functions encoded by the acc operon. These privileged opine-involved activities contribute to the formation of agrobacterial niches in the environment. We found that the expression of the acc operon is induced by agrocinopines and also by limitation of phosphate. The main promoter is present in front of the first gene, accR, which codes for a repressor. This operon structure enables efficient repression when opine levels are low. The promoter contains two putative operators, one overlapping the -10 sequence and the other in the further upstream from it; two partly overlapped putative pho boxes between the two operators; and two consecutive transcription start sites. DNA fragments containing either of the operators bound purified repressor AccR in the absence of agrocinopines but not in the presence of the opines, demonstrating the on-off switch of the promoter. Induction of the acc operon can occur under low-phosphate conditions in the absence of agrocinopines and further increases when the opines also are present. Such opine-phosphate dual regulatory system of the operon may ensure maximum utilization of agrocinopines when available and thereby increase the chances of agrobacterial survival in the highly competitive environment with limited general food sources.


Hypoxia, Hypoxia-inducible Transcription Factors, and Renal Cancer.
Renal cancer is a common urologic malignancy, and therapeutic options for metastatic disease are limited. Most clear cell renal cell carcinomas (ccRCC) are associated with loss of von Hippel-Lindau tumor suppressor (pVHL) function and deregulation of hypoxia pathways. This review summarizes recent evidence from genetic and biological studies showing that hypoxia and hypoxia-related pathways play critical roles in the development and progress of renal cancer. We used a systematic search for articles using the keywords hypoxia, HIF, renal cancer, and VHL. Identification of the tumor suppressor pVHL has allowed the characterization of important ccRCC-associated pathways. pVHL targets α-subunits of hypoxia-inducible transcription factors (HIF) for proteasomal degradation. The two main HIF-α isoforms have opposing effects on RCC biology, possibly through distinct interactions with additional oncogenes. Furthermore, HIF-1α activity is commonly diminished by chromosomal deletion in ccRCCs, and increased HIF-1 activity reduces tumor burden in xenograft tumor models. Conversely, polymorphisms at the HIF-2α gene locus predispose to the development of ccRCCs, and HIF-2α promotes tumor growth. Genetic studies have revealed a prominent role for chromatin-modifying enzyme genes in ccRCC, and these may further modulate specific aspects of the HIF response. This suggests that, rather than global activation of HIF, specific components of the response are important in promoting kidney cancer. Some of these processes are already targets for current therapeutic strategies, and further dissection of this pathway might yield novel methods of treating RCC. In contrast to many tumor types, HIF-1α and HIF-2α have opposing effects in ccRCC biology, with HIF-1α acting as a tumor suppressor and HIF-2α acting as an oncogene. The overall effect of VHL inactivation will depend on fine-tuning of the HIF response. High levels of hypoxia-inducible transcription factors (HIF) are particularly important in the clear cell type of kidney cancer, in which they are no longer properly regulated by the von Hippel-Lindau protein. The two HIF-α proteins have opposing effects on tumor evolution.


Steps to recovery: body weight-supported treadmill training for critically ill patients: a randomized controlled trial.
Early mobilization has been proven effective for patients in intensive care units (ICUs) to improve functional recovery. However, early mobilization of critically ill, often mechanically ventilated, patients is cumbersome because of the attachment to tubes, drains, monitoring devices and muscle weakness. A mobile treadmill with bodyweight support may help to initiate mobilization earlier and more effectively. The aim of this study is to assess the effectiveness of weight-supported treadmill training in critically ill patients during and after ICU stay on time to independent functional ambulation. In this randomized controlled trial, a custom-built bedside body weight-supported treadmill will be used and evaluated. Patients are included if they have been mechanically ventilated for at least 48 hours, are able to follow instructions, have quadriceps muscle strength of Medical Research Council sum-score 2 (MRC 2) or higher, can sit unsupported and meet the safety criteria for physical exercise. Exclusion criteria are language barriers, no prior walking ability, contraindications for physiotherapy or a neurological condition as reason for ICU admission. We aim to include 88 patients and randomize them into either the intervention or the control group. The intervention group will receive usual care plus bodyweight-supported treadmill training (BWSTT) daily. The BWSSTT consists of walking on a mobile treadmill while supported by a harness. The control group will receive usual care physiotherapy treatment daily consisting of progressive activities such as bed-cycling and active functional training exercises. In both groups, we will aim for a total of 40 minutes of physiotherapy treatment time every day in one or two sessions, as tolerated by the patient. The primary outcome is time to functional ambulation as measured in days, secondary outcomes include walking distance, muscle strength, status of functional mobility and symptoms of post-traumatic stress. All measurements will be done by assessors who are blinded to the intervention on the regular wards until hospital discharge. This will be the first study comparing the effects of BWSTT and conventional physiotherapy for critically ill patients during and after ICU stay. The results of this study contribute to a better understanding of the effectiveness of early physiotherapy interventions for critically ill patients. Dutch Trial Register (NTR) ID: NL6766. Registered at 1 December 2017.


"I don't think people are ready to trust these algorithms at face value": trust and the use of machine learning algorithms in the diagnosis of rare disease.
As the use of AI becomes more pervasive, and computerised systems are used in clinical decision-making, the role of trust in, and the trustworthiness of, AI tools will need to be addressed. Using the case of computational phenotyping to support the diagnosis of rare disease in dysmorphology, this paper explores under what conditions we could place trust in medical AI tools, which employ machine learning. Semi-structured qualitative interviews (n = 20) with stakeholders (clinical geneticists, data scientists, bioinformaticians, industry and patient support group spokespersons) who design and/or work with computational phenotyping (CP) systems. The method of constant comparison was used to analyse the interview data. Interviewees emphasized the importance of establishing trust in the use of CP technology in identifying rare diseases. Trust was formulated in two interrelated ways in these data. First, interviewees talked about the importance of using CP tools within the context of a trust relationship; arguing that patients will need to trust clinicians who use AI tools and that clinicians will need to trust AI developers, if they are to adopt this technology. Second, they described a need to establish trust in the technology itself, or in the knowledge it provides-epistemic trust. Interviewees suggested CP tools used for the diagnosis of rare diseases might be perceived as more trustworthy if the user is able to vouchsafe for the technology's reliability and accuracy and the person using/developing them is trusted. This study suggests we need to take deliberate and meticulous steps to design reliable or confidence-worthy AI systems for use in healthcare. In addition, we need to devise reliable or confidence-worthy processes that would give rise to reliable systems; these could take the form of RCTs and/or systems of accountability transparency and responsibility that would signify the epistemic trustworthiness of these tools. words 294.


Genetic variants associated with lung function: the long life family study.
Reduced forced expiratory volume in 1 second (FEV1) and the ratio of FEV1 to forced vital capacity (FVC) are strong predictors of mortality and lung function is higher among individuals with exceptional longevity. However, genetic factors associated with lung function in individuals with exceptional longevity have not been identified. We conducted a genome wide association study (GWAS) to identify novel genetic variants associated with lung function in the Long Life Family Study (LLFS) (n = 3,899). Replication was performed using data from the CHARGE/SpiroMeta consortia. The association between SNPs and FEV1 and FEV1/FVC was analyzed using a linear mixed effects model adjusted for age, age2, sex, height, field center, ancestry principal components and kinship structure to adjust for family relationships separately for ever smokers and never smokers. In the linkage analysis, we used the residuals of the FEV1 and FEV1/FVC, adjusted for age, sex, height, ancestry principal components (PCs), smoking status, pack-years, and field center. We identified nine SNPs in strong linkage disequilibrium in the CYP2U1 gene to be associated with FEV1 and a novel SNP (rs889574) associated with FEV1/FVC, none of which were replicated in the CHARGE/SpiroMeta consortia. Using linkage analysis, we identified a novel linkage peak in chromosome 2 at 219 cM for FEV1/FVC (LOD: 3.29) and confirmed a previously reported linkage peak in chromosome 6 at 28 cM (LOD: 3.33) for FEV1. Future studies need to identify the rare genetic variants underlying the linkage peak in chromosome 6 for FEV1.


The optical system for the 3.5-m ESO telescope.
The optical design of a mirror telescope with a free aperture of 3.5 m is described. Observations can be made in the Cassegrain (RC) focus f/8, in the direct focus f/3, and in the coudé focus f/30. The results of ray tracing in the form of spot diagrams are dealt with. Some technological problems are discussed.


Dichotic listening in psychotic patients.
Patients recently recovered from schizophrenic or manid-depressive psychoses showed larger ear difference scores than normal controls on a dichotic listening test, i.e. when asked to identify dissimilar words fed synchronously to the two ears. The possible significance of the finding is discussed, in relation both to hemispheric organization and to other aspects of cognitive activity which may underlie performance on the task.


Traumatic homonymous hemianopia.
To describe the characteristics of patients with homonymous hemianopia from traumatic brain injury (TBI) seen in our unit between 1989 and 2004. Only patients with a history of TBI, who had detailed clinical information and results of neuroimaging, were included in our study. Demographic characteristics, clinical features, types of visual field defects, location of lesion and evolution of visual field defects were recorded. Of the 880 patients with homonymous hemianopia seen in our unit, 103 patients (112 with homonymous hemianopia) had TBI (74 men and 29 women, mean age 30.7 (SD 15.3) years). Median time from injury to initial visual field testing was 5 (range 0.5-360) months. In all, 64 (57.1%) patients sustained injuries that were motor vehicle-related; 19 (17%) violence-related; 17 (15.2%) due to falls; and 12 (10.7%) because of other blunt head trauma. Visual field defects included complete homonymous hemianopia in 44 (39.3%) patients and incomplete homonymous hemianopia in 68 (60.7%) patients. The lesion was occipital in 14 (12.5%) patients, associated with optic radiation in 26 (23.2%) and the optic tract in 12 (10.7%), and multiple in 60 (53.6%). Most cases of homonymous hemianopia from TBI were motor vehicle-related. Patients were younger, more often male, and had multiple brain lesions more often than patients with homonymous hemianopia from causes other than TBI. A median delay of 5 months was observed before the documentation of the homonymous hemianopia, which may have a major effect on the success of rehabilitation and driving training in these young patients.


Pitch-interval discrimination and musical expertise: is the semitone a perceptual boundary?
The ability to discriminate pitch changes (or intervals) is foundational for speech and music. In an auditory psychophysical experiment, musicians and non-musicians were tested with fixed- and roving-pitch discrimination tasks to investigate the effects of musical expertise on interval discrimination. The tasks were administered parametrically to assess performance across varying pitch distances between intervals. Both groups showed improvements in fixed-pitch interval discrimination as a function of increasing interval difference. Only musicians showed better roving-pitch interval discrimination as interval differences increased, suggesting that this task was too demanding for non-musicians. Musicians had better interval discrimination than non-musicians across most interval differences in both tasks. Interestingly, musicians exhibited improved interval discrimination starting at interval differences of 100 cents (a semitone in Western music), whereas non-musicians showed enhanced discrimination at interval differences exceeding 125 cents. Although exposure to Western music and speech may help establish a basic interval-discrimination threshold between 100 and 200 cents (intervals that occur often in Western languages and music), musical training presumably enhances auditory processing and reduces this threshold to a semitone. As musical expertise does not decrease this threshold beyond 100 cents, the semitone may represent a musical training-induced intervallic limit to acoustic processing.


Navigating the complexity of the therapeutic and clinical use of photography in psychosocial settings: a review of the literature.
In contrast with the recent increase in the use of photography as a therapeutic tool in psychological settings, lack of clarity exists regarding this intervention modality. While the distinction among Photo Therapy, Therapeutic Photography, and Photo Art Therapy is theoretically established we examined whether this classification is confirmed also in practice. Electronic databases (PsychINFO, PubMed, MEDLINE, and Social Sciences Abstracts-EBSCO) were systematically searched for studies published in the last fifteen years utilizing photographic techniques as the elective method of intervention. A total of 124 articles were identified, of which 21 met the inclusion criteria. Results indicate that photography has been used in health, clinical, social, and community settings. Photovoice, Photographic Self-Portrait, and Photo Presentation are the methods most commonly implemented. Several contributions could be placed in more than one of the above-mentioned models of intervention; suggesting that the criteria to assign articles to each model are difficult to apply when the models are put into practice.


Regulation of large cerebral arteries and cerebral microvascular pressure.
Resistance of large arteries appears to be greater in the cerebral circulation than in other vascular beds. Large arteries contribute importantly to total cerebral vascular resistance and are major determinants of local microvascular pressure. Recent studies have shown that resistance of large arteries and cerebral microvascular pressure are affected by several physiological stimuli, including changes in systemic blood pressure, increases in cerebral metabolism, activity of sympathetic nerves, and humoral stimuli such as circulating vasopressin and angiotensin. Stimuli such as sympathetic stimulation and vasopressin produce selective responses of large arteries and, thereby, regulate microvascular pressure without a significant change in cerebral blood flow. These findings lead to the new hypothesis that the brain may be sensitive to changes in cerebral microvascular pressure, resulting in activation of compensatory neurohumoral mechanisms. Important changes occur in large cerebral arteries under pathophysiological conditions. Chronic hypertension increases resistance of large cerebral arteries, which protects the microcirculation against hypertension. Atherosclerosis potentiates constrictor responses of large cerebral arteries to serotonin and thromboxane, which may contribute to vasospasm and transient ischemic attacks.


Psychometric properties of PedsQL generic core scales for children with functional constipation in the Netherlands.
The aim of the study was to evaluate the psychometric properties of the Dutch translation of the Pediatric Quality of Life Inventory version 4.0 Generic Core Scales (PedsQL) in children with functional constipation (FC). The PedsQL was completed by children with FC ages 5 to 18 years and by the parents of children ages 2 to 18 years. To assess construct validity, all of the parents and children completed the PedsQL Gastrointestinal Symptoms Module (GI module) and children ages 8 to 18 years completed the defecation disorder list. Item response distributions, internal consistency reliability, patient-parent agreement, and discriminating ability were evaluated. Overall, 269 children were enrolled. The PedsQL showed minimal missing responses (self-report: 5.3% with >50% missing, parent report: 2.6% with >50% missing) and achieved satisfactory internal consistency for the total (self-report: α = 0.86, parent report: α = 0.88), physical health (self-report: α = 0.75, parent report α = 0.83), and psychosocial health (self-report: α = 0.80; parent report: α = 0.85) scores. Internal consistency was less convincing for those ages 5 to 7 years (α = 0.53-0.77) than for older individuals. Self-reported health-related quality of life was lower than parent reported, with the exception of children ages 5 to 7 years. Interrater reliability intraclass correlations were 0.52 to 0.61 for the overall population. Correlations among the PedsQL, GI module, and defecation disorder list were consistent with a priori hypotheses and generally supported construct validity. The PedsQL distinguished between a previously published healthy reference sample and children with FC, and between those with mild and severe symptoms (P < 0.05). The self-reported and proxy reported PedsQL demonstrated satisfactory measurement properties in Dutch children with FC ages 8 to 18 years. Further research is needed to establish internal consistency and validity in those ages 5 to 7 years.


Crucial role for central carbon metabolism in the bacterial L-form switch and killing by β-lactam antibiotics.
The peptidoglycan cell wall is an essential structure for the growth of most bacteria. However, many are capable of switching into a wall-deficient L-form state in which they are resistant to antibiotics that target cell wall synthesis under osmoprotective conditions, including host environments. L-form cells may have an important role in chronic or recurrent infections. The cellular pathways involved in switching to and from the L-form state remain poorly understood. This work shows that the lack of a cell wall, or blocking its synthesis with β-lactam antibiotics, results in an increased flux through glycolysis. This leads to the production of reactive oxygen species from the respiratory chain, which prevents L-form growth. Compensating for the metabolic imbalance by slowing down glycolysis, activating gluconeogenesis or depleting oxygen enables L-form growth in Bacillus subtilis, Listeria monocytogenes and Staphylococcus aureus. These effects do not occur in Enterococcus faecium, which lacks the respiratory chain pathway. Our results collectively show that when cell wall synthesis is blocked under aerobic and glycolytic conditions, perturbation of cellular metabolism causes cell death. We provide a mechanistic framework for many anecdotal descriptions of the optimal conditions for L-form growth and non-lytic killing by β-lactam antibiotics.


A preliminary overview of the corticioid Atractiellomycetes (Pucciniomycotina, Basidiomycetes).
The taxonomy of the corticioid fungi from the class Atractiellomycetes (Pucciniomycotina, Basidiomycetes) currently addressed to the genus Helicogloea, is revised based on morphological and nuclear ribosomal DNA (ITS and LSU) data. The genus is restricted to 25 species with semitranslucent, gelatinous basidiocarps lacking differentiated cystidia and clamps on hyphae, of which 11 are described as new to science. The asexual genus Leucogloea is placed as a synonym of Helicogloea s. str. Since the type species of Saccoblastia, S. ovispora, is combined to Helicogloea, a new genus, Saccosoma, is introduced to encompass Saccoblastia farinacea and six related species, one of which is described as new. In contrast to Helicogloea in the strict sense, the basidiocarps of Saccosoma are arid, not gelatinized, and hyphae are clamped. The third lineage of the corticioid Atractiellomycetes is represented by the Bourdotigloea vestita complex. Species of Bourdotigloea are devoid of clamps but often possess well-differentiated cystidia, as well as long, cylindrical-fusiform basidiospores. Bourdotigloea encompasses nine species, of which six are described here as new.


SREB, a GATA transcription factor that directs disparate fates in Blastomyces dermatitidis including morphogenesis and siderophore biosynthesis.
Blastomyces dermatitidis belongs to a group of human pathogenic fungi that exhibit thermal dimorphism. At 22 degrees C, these fungi grow as mold that produce conidia or infectious particles, whereas at 37 degrees C they convert to budding yeast. The ability to switch between these forms is essential for virulence in mammals and may enable these organisms to survive in the soil. To identify genes that regulate this phase transition, we used Agrobacterium tumefaciens to mutagenize B. dermatitidis conidia and screened transformants for defects in morphogenesis. We found that the GATA transcription factor SREB governs multiple fates in B. dermatitidis: phase transition from yeast to mold, cell growth at 22 degrees C, and biosynthesis of siderophores under iron-replete conditions. Insertional and null mutants fail to convert to mold, do not accumulate significant biomass at 22 degrees C, and are unable to suppress siderophore biosynthesis under iron-replete conditions. The defect in morphogenesis in the SREB mutant was independent of exogenous iron concentration, suggesting that SREB promotes the phase transition by altering the expression of genes that are unrelated to siderophore biosynthesis. Using bioinformatic and gene expression analyses, we identified candidate genes with upstream GATA sites whose expression is altered in the null mutant that may be direct or indirect targets of SREB and promote the phase transition. We conclude that SREB functions as a transcription factor that promotes morphogenesis and regulates siderophore biosynthesis. To our knowledge, this is the first gene identified that promotes the conversion from yeast to mold in the dimorphic fungi, and may shed light on environmental persistence of these pathogens.


Tris-[4-(di-methyl-amino)-pyridine][tris(pyra-zol-1-yl)methane]--ruthenium(II) bis-(hexa-fluorido-phosphate) diethyl ether monosolvate.
In the title compound, [Ru(C10H10N6)(C7H10N2)3](PF6)2·C4H10O, the Ru(II) cation is coordinated by one tris-(1-pyrazol-yl)methane (Tpm) and three dimethylaminopyridine (dmap) ligands in a slightly distorted octa-hedral geometry. The asymmetric unit consists of one complex cation, two hexa-fluorido-phosphate anions and one diethyl ether solvent mol-ecule in general positions. Although quite a large number of ruthenium complexes of the facially coordinating tridentate Tpm ligand have been structurally characterized, this is only the second one containing three pyridyl co-ligands. The average Ru-N(Tpm) distance is 2.059 (12) Å, while the average Ru-N(dmap) [dmap = 4-(di-methyl-amino)-pyridine] distance is somewhat longer at 2.108 (13) Å. The orientation of the dmap ligands varies greatly, with dihedral angles between the pyridyl and opposite pyrazolyl rings of 14.3 (2), 23.2 (2) and 61.2 (2)°.


Leukaemia classification: a study of the accuracy of diagnosis in 456 patients.
A panel of five haematologists has examined, without consultation or prior knowledge of the diagnosis, blood films and bone marrow smears from 456 patients with a diagnosis of leukaemia. A diagnostic classification which recognized various subtypes of acute myelogenous leukaemia was used but no attempt was made to subdivide acute lymphoblastic leukaemia. Complete agreement with the initial diagnosis was low (56.4%) and was particularly poor (45.7%) when the patient had one of the forms of acute leukaemia. However, disagreements which would have involved the patient in a change of treatment were unusual (2.0%). We conclude that a high degree of diagnostic agreement for patients with leukaemia is unlikely from morphological classifications alone.


Molecular epidemiology of ampicillin resistance in Salmonella spp. and Escherichia coli from wastewater and clinical specimens.
Molecular epidemiology at local scale in Sicily (Italy) of ampicillin resistance in Salmonella spp. isolates from municipal wastewater (n = 64) and clinical specimens (n = 274) is described in comparison with previously examined Escherichia coli isolates (n = 273) from wastewater. High prevalence of antibiotic resistance (28.9%) with highest resistance rates against ampicillin (22.7%) was observed in E. coli isolates. Different resistance rates were observed in Salmonella according to the serovars, with prevalences of the same order in both wastewater and clinical isolates belonging to the same serovar (e.g., 91.7% ampicillin resistance in wastewater isolates vs. 70.8% in clinical isolates of the Salmonella serovar Typhimurium and 0% ampicillin resistance in both wastewater and clinical isolates of the Salmonella serovar Enteritidis). The beta-lactam resistance gene bla(TEM) was present in both wastewater and clinical Salmonella spp. isolates, with the exception of Salmonella enterica serovar Typhimurium isolates with a typical six-drug resistance pattern AmpChlSulTeStrSp that had the bla(PSE-1) gene. The bla(TEM) gene was present in all the E. coli isolates but one had the bla(SHV) gene. Several E. coli and some Salmonella isolates were positive for class 1 integrons with variable regions of 1.0 or 1.5 kb containing aadA1, dfrA17-aadA5, or dfrA1-aadA1 gene cassettes, whereas Salmonella serovar Typhimurium isolates with the six-drug resistance pattern were positive for both 1.0 and 1.2 kb integrons. Polymerase chain reaction replicon typing demonstrated the presence of multireplicon resistance plasmids in several isolates of E. coli, containing two to four of the replicons IncF, IncI1, IncFIA, and IncFIB, whereas other isolates showed resistance plasmids with only IncF, IncP, or IncK replicons. Replicon IncI1 was detected in one Salmonella isolate, whereas other isolates belonging to different serovars had IncN replicons. Analysis of isolates from wastewater can be a useful epidemiologic tool to monitor the prevalence of antibiotic resistance and genetic elements related to antibiotic resistance in Salmonella clones circulating in the human population.


T-lymphocyte-enriched murine peritoneal exudate cells. II. Genetic control of antigen-induced T-lymphocyte proliferation.
The recent introduction of a reliable, T-lymphocyte proliferation assay, which utilizes thioglycollate-induced, nylon wool column-passed, peritoneal exudate lymphocytes from immune mice (PETLES), allowed us to investigate the genetic control of murine immune responses at the T-lymphocyte level. Examination of the blast cells generated in this population 5 days after stimulation with antigen, revealed that 85% of the cells bore the Thy 1 antigen on their surface, whereas only 5% bore immunoglobulin. Thus, the assay can be considered to measure almost exclusively T-lymphocyte function. This assay was used to examine the T-lymphocyte proliferative responses to seven different antigens: poly(Glu60Ala30Tyr10), poly(Glu58Lys38Tyr4), poly-(Tyr,Glu)-poly-D,L-Ala--poly-Lys, poly-(Phe,Glu)-poly-D,L-Ala--poly-Lys, staphylococcal nuclease, lactate dehydrogenase H4, and the BALB/c IgA myeloma protein, TEPC-15. PETLES from a large number of different inbred mouse strains, including H-2 congenic resistant lines and H-2 recombinants, were studied. The strains could be classified as high responders, low responders, or nonresponders to a particular antigen as judged by the magnitude of the T-lymphocyte proliferative response. In every case but one this classification corresponded to the responder status given the strain based on its ability to mount an in vivo antibody response to the same antigen. For two of the antigens, poly-(Tyr,Glu)-poly-D,L-Ala--poly-Lys and TEPC-15, the immune response genes controlling the T-lymphocyte proliferative response were mapped to the K region or I-A subregion of the major histocompatibility complex, as had previously been shown for the control of the antibody responses to these antigens. This tight linkage of the two phenotypic responses very strongly suggests that the same immune response gene controls the expression of both the proliferative and antibody responses. Since there is essentially no contribution from B lymphocytes in the T-lymphocyte proliferation assay, it seems reasonable to conclude that none of the seven immune response genes studied are expressed solely in B lymphocytes.