Assessing the impacts of agricultural intensification on biodiversity: a British perspective.
Agricultural intensification is best considered as the level of human appropriation of terrestrial net primary production. The global value is set to increase from 30%, increasing pressures on biodiversity. The pressures can be classified in terms of spatial scale, i.e. land cover, landscape management and crop management. Different lowland agricultural landscapes in Great Britain show differences among these pressures when habitat diversity and nutrient surplus are used as indicators. Eutrophication of plants was correlated to N surplus, and species richness of plants correlated with broad habitat diversity. Bird species diversity only correlated with habitat diversity when the diversity of different agricultural habitats was taken into account. The pressures of agricultural change may be reduced by minimizing loss of large habitats, minimizing permanent loss of agricultural land, maintaining habitat diversity in agricultural landscapes in order to provide ecosystem services, and minimizing pollution from nutrients and pesticides from the crops themselves. While these pressures could potentially be quantified using an internationally consistent set of indicators, their impacts would need to be assessed using a much larger number of locally applicable biodiversity indicators.


The return to sports activity after conservative or surgical treatment in athletes with lumbar disc herniation.
The purpose of this study was to clarify the efficacy of conservative or surgical treatment in athletes with lumbar disc herniation by reviewing the literature. A search of the literature was performed through PubMed, seeking studies published from 1990 to 2009 with regard to the outcomes of conservative or surgical treatment of lumbar disc herniation in athletes. The percentage of athletes who returned to their original levels of sports activities and the period until their return after the start of treatment were assessed. One prospective and six retrospective studies were identified. One study was for conservative treatment, three for microdiscectomy, and the remaining three for percutaneous discectomy. The respective percentages of athletes who returned to original levels of sports activities (the period until their return) were 78.9% (4.7 mos), 85.1% (5.2-5.8 mos), and 69.9% (7.0 wks to 12 mos). The efficacy of conservative treatment and microdiscectomy, but not percutaneous discectomy, was comparable. No studies were found available for conventional open discectomy, percutaneous laser discectomy, or microendoscopic discectomy. It was revealed that the outcomes of conservative treatment or microdiscectomy in athletes with lumbar disc herniation seemed to be satisfactory in terms of their ability to return the injured athletes to their original levels of sports activities.


Children's intellectual function in relation to arsenic exposure.
Very little evidence exists concerning the possible impairment of children's intellectual function in relation to arsenic exposure in utero and during childhood. We conducted a cross-sectional study among 351 children age 5 to 15 years who were selected from a source population of 7683 people in West Bengal, India, in 2001-2003. Intellectual function was assessed with 6 subtests from the Wechsler Intelligence Scale for Children as well as with the Total Sentence Recall test, the Colored Progressive Matrices test, and a pegboard test. Arsenic in urine and lifetime water sources (including during the pregnancy period) were assessed using measurements of samples from 409 wells. The test scores were analyzed with linear regression analyses based on the method of generalized estimating equations incorporating relevant covariates. Stratifying urinary arsenic concentrations into tertiles, we found associations between arsenic and reductions in the adjusted scores of the vocabulary test (0, -0.14, -0.28; P for trend = 0.02), the object assembly test (0, -0.16, -0.24; P for trend = 0.03), and the picture completion test (0, -0.15, -0.26; P for trend = 0.02). These findings correspond to relative declines of 12% (95% confidence interval =0.4% to 24%) in the vocabulary test, 21% (-0.8% to 42%) in the object assembly test, and of 13% (0.3% to 24%) in the picture completion test in the upper urinary arsenic tertile. However, we did not find evidence of an association between test results and arsenic water concentrations during pregnancy or childhood. Current arsenic concentrations in urine, which reflect all sources of recent exposure, including water and food, were associated with small decrements in intellectual testing in school-aged children in West Bengal. We did not see associations between long-term water arsenic concentrations and intellectual function.


Benchtop fabrication of PDMS microstructures by an unconventional photolithographic method.
Poly(dimethylsiloxane) (PDMS) microstructures have been widely used in bio-microelectromechanical systems (bio-MEMS) for various types of analytical, diagnostic and therapeutic applications. However, PDMS-based soft lithographic techniques still use conventional microfabrication processes to generate a master mold, which requires access to clean room facilities and costly equipment. With the increasing use of these systems in various fields, the development of benchtop systems for fabricating microdevices is emerging as an important challenge in their widespread use. Here we demonstrate a simple, low-cost and rapid method to fabricate PDMS microstructures by using micropatterned poly(ethylene glycol) diacrylate (PEGDA) master molds. In this method, PEGDA microstructures were patterned on a glass substrate by photolithography under ambient conditions and by using simple tools. The resulting PEGDA structures were subsequently used to generate PDMS microstructures by standard molding in a reproducible and repeatable manner. The thickness of the PEGDA microstructures was controllable from 15 to 300 µm by using commonly available spacer materials. We also demonstrate the use of this method to fabricate microfluidic channels capable of generating concentration gradients. In addition, we fabricated PEGDA microstructures by photolithography from the light generated from commonly available laminar cell culture hood. These data suggest that this approach could be beneficial for fabricating low-cost PDMS-based microdevices in resource limited settings.


Interactions of orthosteric and allosteric ligands with [3H]dimethyl-W84 at the common allosteric site of muscarinic M2 receptors.
An optimized assay for the binding of [3H]dimethyl-W84 to its allosteric site on M2 muscarinic receptors has been used to directly measure the affinities of allosteric ligands. Their potencies agree with those deduced indirectly by their modulation of the equilibrium binding and kinetics of [3H]N-methylscopolamine ([3H]NMS) binding to the orthosteric site. The affinities and cooperativities of orthosteric antagonists with [3H]dimethyl-W84 have also been quantitated. These affinities agree with those measured directly in a competition assay using [3H]NMS. All these data are compatible with the predictions of the allosteric ternary complex model. The association and dissociation kinetics of [3H]dimethyl-W84 are rapid but the estimate of its association rate constant is nevertheless comparable with that found for the orthosteric radioligand, [3H]NMS. This is unexpected, given that the allosteric site to which [3H]dimethyl-W84 binds is thought to be located on the external face of the receptor and above the [3H]NMS binding site that is buried within the transmembrane helices. The atypical allosteric ligands tacrine and 4,4'-bis-[(2,6-dichloro-benzyloxy-imino)-methyl]-1,1'-propane-1,3-diyl-bis-pyridinium dibromide (Duo3) inhibit [3H]dimethyl-W84 binding with the same potencies and comparably steep slope factors as found for inhibition of [3H]NMS binding. Tacrine and Duo3 decrease [3H]dimethyl-W84 affinity, not the number of binding sites. It is suggested that these atypical ligands either bind to the two known spatially separated allosteric sites on muscarinic receptors with positive cooperativity or their binding to the common allosteric site modulates receptor-receptor interactions such that homotropic positive cooperativity within a dimer or higher oligomer is generated.


Evolution of acuteness in pathogen metapopulations: conflicts between "classical" and invasion-persistence trade-offs.
Classical life-history theory predicts that acute, immunizing pathogens should maximize between-host transmission. When such pathogens induce violent epidemic outbreaks, however, a pathogen's short-term advantage at invasion may come at the expense of its ability to persist in the population over the long term. Here, we seek to understand how the classical and invasion-persistence trade-offs interact to shape pathogen life-history evolution as a function of the size and structure of the host population. We develop an individual-based infection model at three distinct levels of organization: within an individual host, among hosts within a local population, and among local populations within a metapopulation. We find a continuum of evolutionarily stable pathogen strategies. At one end of the spectrum-in large well-mixed populations-pathogens evolve to greater acuteness to maximize between-host transmission: the classical trade-off theory applies in this regime. At the other end of the spectrum-when the host population is broken into many small patches-selection favors less acute pathogens, which persist longer within a patch and thereby achieve enhanced between-patch transmission: the invasion-persistence tradeoff dominates in this regime. Between these extremes, we explore the effects of the size and structure of the host population in determining pathogen strategy. In general, pathogen strategies respond to evolutionary pressures arising at both scales.


Parental Authoritativeness, Social Support and Help-seeking for Mental Health Problems in Adolescents.
Adolescents experience high rates of mental health problems but are reluctant to seek professional help. Parents play an integral role in the help seeking process for their adolescent children. Parental authoritativeness and support have been identified as contributing to better mental health outcomes and a reduction in help seeking barriers in their children. The current study examined the influence of parental authoritativeness and support on help seeking intentions and behaviors in 1582 students (49% female) in 17 schools (mean age = 17.7, range 16-18). All data was available for 1032 participants across two time points collected one year apart. Concurrent indices of positive parenting were associated with greater help seeking intentions from professional sources, even when gender and psychological distress were controlled. Parental authoritativeness and parental support did not predict actual help seeking assessed one year later. The study highlights the potential role of parents in influencing help seeking and suggests further research is needed on other parent variables and the social antecedents to help-seeking.


A model of angiogenesis in the mouse cornea.
The study of angiogenesis depends on reliable and reproducible models for the stimulation of a neovascular response. The purpose of this research was to develop such a model of angiogenesis in the mouse cornea. Uniformly sized Hydron pellets containing either basic fibroblast growth factor (bFGF) or vascular endothelial growth factor (VEGF) and sucralfate were prepared and implanted into the stroma mouse cornea adjacent to the temporal limbus. Neovascularization of the corneal stroma began on day 3 and was sustained through day 8. The bFGF-induced neovascularization was consistent and dose dependent in C57B1/6, as well as in severe combined immune deficient, beige natural killer cell-deficient, and nude mouse strains. Biomicroscopic and histologic examination of bFGF- and VEGF-induced angiogenesis was notable for the absence of corneal edema or substantial inflammation. This noninflammatory model of corneal neovascularization is especially advantageous because it is reproducible, economical, and facilitates investigation of angiogenesis in various murine tumor models as well as in genetically defined murine strains.


Progress in cerebral transplantation of expanded neuronal stem cells.
Given the success and limitations of human fetal primary neural tissue transplantation, neuronal stem cells (NSCs) that can be adequately expanded in culture have been the focus of numerous attempts to develop a superior source of replacement cells for restorative neurosurgery. To clarify recent progress toward this goal, the transplantation into the adult brain of NSCs, expanded in vitro before grafting, was reviewed. Neuronal stem cells can be expanded from a variety of sources, including embryos, fetuses, adult bone marrow, and adult brain tissue. Recent investigations of each of these expanded stem cell types have generated a large body of information along with a great number of unanswered questions regarding the ability of these cells to replace damaged neurons. Expanded NSCs offer many advantages over their primary tissue predecessors, but also may exhibit different functional abilities as grafted cells. Because expanded NSCs will most likely ultimately replace primary tissue grafting in clinical trials, this review was undertaken to focus solely on this distinct body of work and to summarize clearly the existing preclinical data regarding the in vivo successes, limits, and unknowns of using each expanded NSC type when transplanted into the adult brain. Embryonic stem cell-derived cells have demonstrated appropriate neuronal phenotypes after transplantation into nonneurogenic areas of the adult brain. Understanding the mechanisms responsible for this may lead to similar success with less studied adult neuronal progenitor cells, which offer the potential for autologous NSC transplantation with less risk of tumorigenesis.


The endemic plants of Mozambique: diversity and conservation status.
An annotated checklist of the 271 strict-endemic taxa (235 species) and 387 near-endemic taxa (337 species) of vascular plants in Mozambique is provided. Together, these taxa constitute c. 9.3% of the total currently known flora of Mozambique and include five strict-endemic genera (Baptorhachis, Emicocarpus, Gyrodoma, Icuria and Micklethwaitia) and two near-endemic genera (Triceratella and Oligophyton). The mean year of first publication of these taxa is 1959, with a marked increase in description noted following the onset of the two major regional floristic programmes, the "Flora of Tropical East Africa" and "Flora Zambesiaca", and an associated increase in botanical collecting effort. New taxa from Mozambique continue to be described at a significant rate, with 20 novelties described in 2018. Important plant families for endemic and near-endemic taxa include Fabaceae, Rubiaceae and Euphorbiaceae s.s. There is a high congruence between species-rich plant families and endemism with the notable exceptions of the Poaceae, which is the second-most species rich plant family, but outside of the top ten families in terms of endemism, and the Euphorbiaceae, which is the seventh-most species rich plant family, but third in terms of endemism. A wide range of life-forms are represented in the endemic and near-endemic flora, with 49% being herbaceous or having herbaceous forms and 55% being woody or having woody forms. Manica Province is by far the richest locality for near-endemic taxa, highlighting the importance of the cross-border Chimanimani-Nyanga (Manica) Highlands shared with Zimbabwe. A total of 69% of taxa can be assigned to one of four cross-border Centres of Endemism: the Rovuma Centre, the Maputaland Centre sensu lato, and the two mountain blocks, Chimanimani-Nyanga and Mulanje-Namuli-Ribaue. Approximately 50% of taxa have been assessed for their extinction risk and, of these, just over half are globally threatened (57% for strict-endemics), with a further 10% (17% for strict-endemics) currently considered to be Data Deficient, highlighting the urgent need for targeted conservation of Mozambique's unique flora. This dataset will be a key resource for ongoing efforts to identify "Important Plant Areas - IPAs" in Mozambique, and to promote the conservation and sustainable management of these critical sites and species, thus enabling Mozambique to meet its commitments under the Convention on Biological Diversity (CBD).


Compartmentalisation in aquaculture production systems.
Compartmentalisation is a new tool for disease management within a country. In aquaculture, the successful application of compartmentalisation is largely dependent on the system of production and the epidemiology of the disease(s) for which the compartment is being defined. Therefore, compartmentalisation may not be universally applicable across all systems and diseases. The paper examines the implementation of the concept, providing examples of specific industries, and discusses the application of hazard analysis and critical control points (HACCP) to the biosecurity of the system. The role of compartmentalisation in the management of aquaculture disease emergencies is also discussed.


Autocrine IL-10 impairs dendritic cell (DC)-derived immune responses to mycobacterial infection by suppressing DC trafficking to draining lymph nodes and local IL-12 production.
The production of IL-12 by dendritic cells (DC) early in an immune response is considered critical for the polarization of CD4(+) T lymphocyte response towards a Th1 pattern, a key process in the clearance of intracellular pathogens. Infection of bone marrow-derived DC with Mycobacterium bovis Bacillus Calmette Guérin (BCG) induced a concurrent and dose-dependent releaseof IL-10 and IL-12. Here we examined whether the production of IL-10 by DC affected their IL-12 response to mycobacterial infection and the generation of protective immune responses in vivo. Compared to wild-type (WT) DC, DC deficient for IL-10 synthesis (IL-10(-/-)) showed increased IL-12 production in response to BCG infection and CD40 stimuli in vitro. Moreover, when transferred into mice, infected IL-10(-/-) DC were more efficient than WT DC at inducing IFN-gamma production to mycobacterial antigens in the draining lymph nodes (DLN). This effect was associated with increased trafficking of IL-10(-/-) DC to the DLN and enhanced IL-12 production by DC within the DLN. These data show that autocrine IL-10 exerts a dual inhibitory effect on the induction of primary immune responses by DC: first, by down-regulating the migration of infected DC to the DLN and second, by modulating the IL-12 production by DC in the DLN.


DNA polymerase alpha. An immunohistochemical marker for proliferating cells in normal and neoplastic human tissues.
DNA polymerase alpha is a DNA replicating enzyme expressed in all proliferating cells. This nuclear antigen in paraformaldehyde-postfixed frozen sections of normal, benign, and malignant tissues was identified by the peroxidase-antiperoxidase technique with the use of a mouse monoclonal antibody against bovine/human DNA polymerase alpha. The nuclei of normal proliferating cells were positive. Malignant tumors (n = 95) showed a higher proportion of positive nuclei than did low-grade malignant tumors (n = 7) or benign lesions (n = 67). The number of positive nuclei in squamous cell carcinomas (n = 19) was higher than in adenocarcinomas (n = 45). Eight (18%) adenocarcinomas and all five renal cell carcinomas had less than 10% positive cells, whereas in benign tissues, such as pituitary adenomas, a thymoma, reactive lymphoid lesions, and some benign mammary nodules, more than 10% of nuclei were labeled. In addition, foci of proliferating cells were clearly recognized. DNA polymerase alpha is, therefore, an excellent marker of proliferative activity that provides an approach to analyzing tumor cell heterogeneity not only in fully developed neoplasms, but also in their precursor lesions.


Insulitis in type 2 diabetes.
Islets of patients with type 2 diabetes have the feature of an inflammatory process reflected by the presence of cytokines, immune cells, beta-cell apoptosis, amyloid deposits and fibrosis. Indeed, beta-cells from patients with type 2 diabetes display inflammatory markers, including increased interleukin (IL)-1 beta expression. Furthermore, increased islet-associated macrophages are observed in human type 2 diabetic patients and in most animal models of diabetes. Importantly, increased numbers of macrophages are detectable very early in high fat-fed mice islets, before the onset of diabetes. These immune cells are most likely attracted by islet-derived chemokines, produced in response to metabolic stress, and under the control of IL-1 beta. It follows that modulation of intra-islet inflammatory mediators, in particular IL-1 beta, may prevent insulitis in type 2 diabetes and therefore presents itself as a possible causal therapy with disease-modifying potential.


An outbreak of thyroid hyperplasia (goiter) with high mortality in budgerigars (Melopsittacus undulatus).
An outbreak of goiter with high morbidity and mortality in a flock of budgerigars (Melopsittacus undulatus) in California is described. Forty-five out of 400 adult birds exhibited signs of illness, weight loss, and enlargement in the crop area; 15 of the 45 birds died over a 2-3-month period. Diet consisted of a commercial mixture with the addition of broccoli, whole oats, and carrots, but no minerals or supplements. Six budgerigars were subjected to necropsy; all 6 birds had severely enlarged thyroid glands. Thyroid follicular hyperplasia was histologically observed in all birds examined, while granulomatous thyroiditis and microfollicular adenoma were observed in 2 birds, respectively. Virological, bacteriological, parasitological, and heavy metal analyses were negative or within normal limits. The total iodine in the thyroid glands of affected birds was measured by inductively coupled plasma-mass spectrometry. Following iodine supplementation and removal of broccoli from the diet, the owner reported weight gain and a reduced death rate among clinically affected birds; no additional birds became sick. The presence of broccoli with its iodine-binding ability and the complete lack of added minerals in the diet of these animals were thought to be the predisposing factors for the outbreak in the present study. Outbreaks of goiter accompanied by high mortality are rare in any species and, to the best of the authors' knowledge, have not been described previously in any avian species. Recognition of this condition may help improve medical, welfare, and trade standards concerning this species.


Probing the Role of Active Site Water in the Sesquiterpene Cyclization Reaction Catalyzed by Aristolochene Synthase.
Aristolochene synthase (ATAS) is a high-fidelity terpenoid cyclase that converts farnesyl diphosphate exclusively into the bicyclic hydrocarbon aristolochene. Previously determined crystal structures of ATAS complexes revealed trapped active site water molecules that could potentially interact with catalytic intermediates: water "w" hydrogen bonds with S303 and N299, water molecules "w1" and "w2" hydrogen bond with Q151, and a fourth water molecule coordinates to the Mg(2+)C ion. There is no obvious role for water in the ATAS mechanism because the enzyme exclusively generates a hydrocarbon product. Thus, these water molecules are tightly controlled so that they cannot react with carbocation intermediates. Steady-state kinetics and product distribution analyses of eight ATAS mutants designed to perturb interactions with active site water molecules (S303A, S303H, S303D, N299A, N299L, N299A/S303A, Q151H, and Q151E) indicate relatively modest effects on catalysis but significant effects on sesquiterpene product distributions. X-ray crystal structures of S303A, N299A, N299A/S303A, and Q151H mutants reveal minimal perturbation of active site solvent structure. Seven of the eight mutants generate farnesol and nerolidol, possibly resulting from addition of the Mg(2+)C-bound water molecule to the initially formed farnesyl cation, but no products are generated that would suggest enhanced reactivity of other active site water molecules. However, intermediate germacrene A tends to accumulate in these mutants. Thus, apart from the possible reactivity of Mg(2+)C-bound water, active site water molecules in ATAS are not directly involved in the chemistry of catalysis but instead contribute to the template that governs the conformation of the flexible substrate and carbocation intermediates.


Heterogeneity of host immunological risk factors in patients with aggressive periodontitis.
The pathogenesis of early-onset periodontitis (EOP) can be explained by various host risk factors. Previous studies have focused on a single (among many possible) immunological risk factor and the association among the factors has not been assessed. We comprehensively investigated the associations among multiple host immunological risk factors in EOP patients to further elucidate their role in the pathogenesis of EOP. Sixty-eight EOP patients (50 generalized EOP, 18 localized EOP), 51 EOP-suspected patients (S-EOP), 43 adult periodontitis (AP) patients, and 36 periodontally healthy subjects (HS) participated in this cross-sectional study. We examined peripheral neutrophil functions, phenotypic and functional characterization of peripheral lymphocytes (lymphocyte subsets, T-cell proliferative activity), cytokine productivity (interleukin [IL]-1, IL-2, tumor necrosis factor [TNF]-alpha, interferon [IFN]-gamma, IL-4 and IL-6), serum immunoglobulin G (IgG) antibody titers against 12 periodontal bacteria, and HLA class II genotypes. G-EOP, S-EOP, and AP patient groups showed significantly lower percentages of pan T cells and CD8-positive cells (P < 0.02) compared with the HS group. L-EOP patients showed depressed IL-4 and TNF-alpha productivity compared with the HS group (P < 0.02). The EOP group showed significantly elevated antibody levels against Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, and Fusobacterium nucleatum compared with the HS group (P < 0.05). The frequency with DQB1*0503 was significantly higher in the EOP patient group than the HS group (P = 0.045) due to the higher frequency in L-EOP patients than the HS group (P = 0.035). There were wide interindividual variations in each of the tests among patient and HS groups; however, EOP patients showed wider intradiagnostic group variations in certain host defensive cell functions than the other groups. There were some EOP patients who showed extremely low or high values in some tests; the EOP patients could be further divided into subgroups according to their host defensive and immunological profiles. However, there was heterogeneity in some of the other host immunological tests even in the subgroups. The association of host immunological risk factors in EOP patients is widely varied and more complex than previously thought. These results indicate the difficulty of explaining the pathogenesis of EOP based on a single host risk factor and also emphasize the importance of critical assessment of not only EOP patient groups, but also individual patients.


Suprapubic laparoscopic cholecystectomy: technique and preliminary results.
The minimally invasive abdominal surgery has evolved to reduce portals, culminating with a single incision and natural orifice operation. However, these methods are still expensive, difficult to implement and with questionable aesthetic results. To present the standardization and preliminary results of a technique for performing laparoscopic suprapubic access by the principle which was called the Supra Pubic Endoscopic Surgery for cholecystectomy. The average body mass index of patients, the mean operative time, clinical data of the postoperative complications and quality of life were prospectively studied. The operation incisions consisted of: A) umbilical for instrumental dissection and clipping; B) in the right groin for handling and gallbladder gripping; C) suprapubic for the camera. With the patient in reverse Trendelenburg and left lateral decubitus, the operation flew by the camera trocar in C, proceeding with dissection and isolation of the biliary pedicle, identification of cystic duct and artery, with usual instrumentation. Transcystic intraoperative cholangiography was performed in all cases in which there were indications. The procedure was completed with clipping and sectioning of the cystic duct and artery, retrograde resection of the gallbladder and extracting it by the umbilical trocar incision under direct vision. Thirty patients undergone this surgical procedure between March and June 2012 and were evaluated. The mean age was 40.7 years and the indications were typical biliary colic in 18 cases (60 %), cholecystitis in five cases (16.6 %), biliary pancreatitis in one case (3.3%); polyp in three cases (10%) and obstructive jaundice at three cases (10%). The average body mass index was 27.8 (23.1-35.1) and surgical time ranged between 24 and 70 minutes. The technique proved to be feasible and safe , with no significant complications, and satisfactory cosmetic results.


Effects of neonatal rat Borna disease virus (BDV) infection on the postnatal development of the brain monoaminergic systems.
Effects of neonatal Borna disease virus infection (BDV) on the postnatal development of brain monoaminergic systems in rats were studied. Tissue content of norepinephrine (NE), dopamine (DA) and its metabolite, 3,4-dihydroxyphenol acetic acid (DOPAC), and serotonin (5-HT) and its metabolite, 5-hydroxyindole-3-acetic acid (5-HIAA) were assayed by means of HPLC-EC in frontal cortex, cerebellum, hippocampus, hypothalamus and striatum of neonatally BDV-infected and sham-inoculated male Lewis rats of 8, 14, 21, 60 and 90 days of age. Both NE and 5-HT concentrations were significantly affected by neonatal BDV infection. The cortical and cerebellar levels of NE and 5-HT were significantly greater in BDV-infected rats than control animals at postnatal days (PND) 60 and 90. Tissue content of NE in hippocampus was unaffected. In hippocampus, neonatally BDV-infected rats had lower 5-HT levels at PND 8 and significantly elevated levels at PND 21 and onwards. Neither striatal levels of 5-HT nor hypothalamic levels of 5-HT and NE were affected by neonatal BDV infection, suggesting that the monoamine systems in the prenatally maturing brain regions are less sensitive to effects of neonatal viral infection. 5-HIAA/5-HT ratio was not altered in BDV-infected rats indicating no changes in the 5-HT turnover in the brain regions damaged by the virus. Neither DA nor DOPAC/DA ratio was affected by neonatal BDV infection in any of the brain regions examined. The present data demonstrate significant and specific alterations in monoaminergic systems in neonatally BDV-infected rats. This pattern of changes is consistent with the previously reported behavioral abnormalities resulting from neonatal BDV infection.


Evaluation of a focused virtual library of heterobifunctional ligands for Clostridium difficile toxins.
A focused library of virtual heterobifunctional ligands was generated in silico and a set of ligands with recombined fragments was synthesized and evaluated for binding to Clostridium difficile toxins. The position of the trisaccharide fragment was used as a reference for filtering docked poses during virtual screening to match the trisaccharide ligand in a crystal structure. The peptoid, a diversity fragment probing the protein surface area adjacent to a known binding site, was generated by a multi-component Ugi reaction. Our approach combines modular fragment-based design with in silico screening of synthetically feasible compounds and lays the groundwork for future efforts in development of composite bifunctional ligands for large clostridial toxins.


Somatic Coliphage Profiles of Produce and Environmental Samples from Farms in Northern México.
Somatic coliphages were quantified in 459 produce and environmental samples from 11 farms in Northern Mexico to compare amounts of somatic coliphages among different types of fresh produce and environmental samples across the production steps on farms. Rinsates from cantaloupe melons, jalapeño peppers, tomatoes, and the hands of workers, soil, and water were collected during 2011-2012 at four successive steps on each farm, from the field before harvest through the packing facility, and assayed by FastPhage MPN Quanti-tray method. Cantaloupe farm samples contained more coliphages than jalapeño or tomato (p range <0.01-0.03). Across production steps, jalapeños had higher coliphage percentages before harvest than during packing (p = 0.03), while tomatoes had higher coliphage concentrations at packing than all preceding production steps (p range <0.01-0.02). These findings support the use of targeted produce-specific interventions at multiple points in the process of growing and packing produce to reduce the risk of enteric virus contamination and improve food safety during fruit and vegetable production.


Ammonium as a signal for physiological and morphological responses in plants.
Ammonium is a major inorganic nitrogen source for plants. At low external supplies, ammonium promotes plant growth, while at high external supplies it causes toxicity. Ammonium triggers rapid changes in cytosolic pH, in gene expression, and in post-translational modifications of proteins, leading to apoplastic acidification, co-ordinated ammonium uptake, enhanced ammonium assimilation, altered oxidative and phytohormonal status, and reshaped root system architecture. Some of these responses are dependent on AMT-type ammonium transporters and are not linked to a nutritional effect, indicating that ammonium is perceived as a signaling molecule by plant cells. This review summarizes current knowledge of ammonium-triggered physiological and morphological responses and highlights existing and putative mechanisms mediating ammonium signaling and sensing events in plants. We put forward the hypothesis that sensing of ammonium takes place at multiple steps along its transport, storage, and assimilation pathways.


A prognostic model for ovarian cancer.
About 6000 women in the United Kingdom develop ovarian cancer each year and about two-thirds of the women will die from the disease. Establishing the prognosis of a woman with ovarian cancer is an important part of her evaluation and treatment. Prognostic models and indices in ovarian cancer should be developed using large databases and, ideally, with complete information on both prognostic indicators and long-term outcome. We developed a prognostic model using Cox regression and multiple imputation from 1189 primary cases of epithelial ovarian cancer (with median follow-up of 4.6 years). We found that the significant (P< or = 0.05) prognostic factors for overall survival were age at diagnosis, FIGO stage, grade of tumour, histology (mixed mesodermal, clear cell and endometrioid versus serous papillary), the presence or absence of ascites, albumin, alkaline phosphatase, performance status on the ZUBROD-ECOG-WHO scale, and debulking of the tumour. This model is consistent with other models in the ovarian cancer literature; it has better predictive ability and, after simplification and validation, could be used in clinical practice.


The Notch pathway controls fibrotic and regenerative repair in the adult heart.
In the adult heart, Notch signalling regulates the response to injury. Notch inhibition leads to increased cardiomyocyte apoptosis, and exacerbates the development of cardiac hypertrophy and fibrosis. The role of Notch in the mesenchymal stromal cell fraction, which contains cardiac fibroblasts and cardiac precursor cells, is, however, largely unknown. In the present study, we evaluate, therefore, whether forced activation of the Notch pathway in mesenchymal stromal cells regulates pathological cardiac remodelling. We generated transgenic mice overexpressing the Notch ligand Jagged1 on the surface of cardiomyocytes to activate Notch signalling in adjacent myocyte and non-myocyte cells. In neonatal transgenic mice, activated Notch sustained cardiac precursor and myocyte proliferation after birth, and led to increased numbers of cardiac myocytes in adult mice. In the adult heart under pressure overload, Notch inhibited the development of cardiomyocyte hypertrophy and transforming growth factor-β/connective tissue growth factor-mediated cardiac fibrosis. Most importantly, Notch activation in the stressed adult heart reduced the proliferation of myofibroblasts and stimulated the expansion of stem cell antigen-1-positive cells, and in particular of Nkx2.5-positive cardiac precursor cells. We conclude that Notch is pivotal in the healing process of the injured heart. Specifically, Notch regulates key cellular mechanisms in the mesenchymal stromal cell population, and thereby controls the balance between fibrotic and regenerative repair in the adult heart. Altogether, these findings indicate that Notch represents a unique therapeutic target for inducing regeneration in the adult heart via mobilization of cardiac precursor cells.


Specific binding of cardiac glycoside drugs and endogenous digitalis-like substances to particulate membrane fractions from human placenta.
We studied the characteristics of binding of cardiac glycosides to particulate membrane fractions from human placenta, to demonstrate that placental tissue is a suitable source of receptors for digitalis drugs. Moreover, we performed preliminary experiments with 125I-labeled digoxin and placental particulates to develop a radioreceptor assay for measurement of endogenous substances with activity similar to cardiac glycoside drugs (EDLS). Placental membrane fractions were incubated with [3H]ouabain (10 nmol/L) or 125I-labeled digoxin (50 pmol/L). With both ligands, binding followed a pseudo-first-order reaction kinetics and was saturable. Scatchard analysis revealed a single class of sites [for ouabain, KD = 20.2 +/- 5.8 nmol/L (mean +/- SEM), Bmax = 3.1 +/- 0.9 nmol per gram of protein; for digoxin, KD = 29.7 +/- 1.9 nmol/L, Bmax = 24.3 +/- 1.1 nmol per gram of protein]. As expected, digoxin was less potent than ouabain in displacing both tracers from digitalis drugs receptors; progesterone, cortisone, digitoxose, furosemide, bumetanide, and propranolol had no or little effect. Specific 125I-labeled digoxin binding was competitively inhibited by plasma and (or) urine extracts from newborns, adults, pregnant women, and patients with renal insufficiency. Inhibition of binding and volume of plasma and urine assayed were linearly related. These findings support the hypothesis that cardiac glycosides and EDLS can interact with the human placenta and suggest placental tissue to be a suitable source of receptors for cardiac glycosides.


Winner of the "New Investigator Award" at the European Society of Hyperthermia Oncology Meeting 2007. The HYPERcollar: a novel applicator for hyperthermia in the head and neck.
The purpose of this work was to define all features, and show the potential, of the novel HYPERcollar applicator system for hyperthermia treatments in the head and neck region. The HYPERcollar applicator consists of (1) an antenna ring, (2) a waterbolus system and (3) a positioning system. The specific absorption rate (SAR) profile of this applicator was investigated by performing infra-red measurements in a cylindrical phantom. Mandatory patient-specific treatment planning was performed as an object lesson to a patient with a laryngeal tumour and an artificial lymph node metastasis. Comfort tests with healthy volunteers have revealed that the applicator provides sufficient comfort to maintain in treatment position for an hour: the standard hyperthermia treatment duration in our centre. By phantom measurements, we established that a central focus in the neck can be obtained, with 50% iso-SAR lengths of 3.5 cm in transversal directions (x/y) and 9-11 cm in the axial direction (z). Using treatment planning by detailed electromagnetic simulations, we showed that the SAR pattern can be optimised to enable simultaneous encompassing of a primary laryngeal tumour and a lymph node metastasis at the 25% iso-SAR level. This study shows that the applicator enables a good control, and sufficient possibilities for optimisation, of the SAR pattern. In an ongoing clinical feasibility study, we will investigate the possibilities of heating various target regions in the neck with this apparatus.


Influence of Rule- and Reward-based Strategies on Inferences of Serial Order by Monkeys.
Knowledge of transitive relationships between items can contribute to learning the order of a set of stimuli from pairwise comparisons. However, cognitive mechanisms of transitive inferences based on rank order remain unclear, as are relative contributions of reward associations and rule-based inference. To explore these issues, we created a conflict between rule- and reward-based learning during a serial ordering task. Rhesus macaques learned two lists, each containing five stimuli that were trained exclusively with adjacent pairs. Selection of the higher-ranked item resulted in rewards. "Small reward" lists yielded two drops of fluid reward, whereas "large reward" lists yielded five drops. Following training of adjacent pairs, monkeys were tested on novels pairs. One item was selected from each list, such that a ranking rule could conflict with preferences for large rewards. Differences between the corresponding reward magnitudes had a strong influence on accuracy, but we also observed a symbolic distance effect. That provided evidence of a rule-based influence on decisions. RT comparisons suggested a conflict between rule- and reward-based processes. We conclude that performance reflects the contributions of two strategies and that a model-based strategy is employed in the face of a strong countervailing reward incentive.


Modeling gating charge and voltage changes in response to charge separation in membrane proteins.
Measurements of voltage changes in response to charge separation within membrane proteins can offer fundamental information on mechanisms of charge transport and displacement processes. A recent example is provided by studies of cytochrome c oxidase. However, the interpretation of the observed voltage changes in terms of the number of charge equivalents and transfer distances is far from being trivial or unique. Using continuum approaches to describe the voltage generation may involve significant uncertainties and reliable microscopic simulations are not yet available. Here, we attempt to solve this problem by using a coarse-grained model of membrane proteins, which includes an explicit description of the membrane, the electrolytes, and the electrodes. The model evaluates the gating charges and the electrode potentials (c.f. measured voltage) upon charge transfer within the protein. The accuracy of the model is evaluated by a comparison of measured voltage changes associated with electron and proton transfer in bacterial photosynthetic reaction centers to those calculated using our coarse-grained model. The calculations reproduce the experimental observations and thus indicate that the method is of general use. Interestingly, it is found that charge-separation processes with different spatial directions (but the same distance perpendicular to the membrane) can give similar observed voltage changes, which indicates that caution should be exercised when using simplified interpretation of the relationship between charge displacement and voltage changes.


Separate visual representations in the planning and control of action.
Evidence for a dichotomy between the planning of an action and its on-line control in humans is reviewed. This evidence suggests that planning and control each serve a specialized purpose utilizing distinct visual representations. Evidence from behavioral studies suggests that planning is influenced by a large array of visual and cognitive information, whereas control is influenced solely by the spatial characteristics of the target, including such things as its size, shape, orientation, and so forth. Evidence from brain imaging and neuropsychology suggests that planning and control are subserved by separate visual centers in the posterior parietal lobes, each constituting part of a larger network for planning and control. Planning appears to rely on phylogenetically newer regions in the inferior parietal lobe, along with the frontal lobes and basal ganglia, whereas control appears to rely on older regions in the superior parietal lobe, along with the cerebellum.


Tuning the hydrogen evolution activity of β-Mo2C nanoparticles via control of their growth conditions.
The use of water electrocatalysis for hydrogen production is a promising, sustainable and greenhouse-gas-free process to develop disruptive renewable energy technologies. Transition metal carbides, in particular β-phase Mo2C, are garnering increased attention as hydrogen evolution reaction (HER) catalysts due to their favourable synthesis conditions, stability and high catalytic efficiency. We use a thermodynamic approach in conjunction with density functional theory and a kinetic model of exchange current density to systematically study the HER activity of β-Mo2C under different experimental conditions. We show that the (011) surface has the highest HER activity, which is rationalized by its lack of strong Mo-based hydrogen adsorption sites. Thus, the HER efficiency of β-Mo2C can be tuned using nanoparticles (NPs) that expose larger fractions of this termination. We give definite maps between NP morphologies and experimental synthesis conditions, and show that the control of the carbon chemical potential during synthesis can expose up to 90% of the (011) surface, while ambient H2 has little effect on the NP morphology. The "volcano" plot shows that under these optimum conditions, the NP exchange current density is ∼10-5 A cm-2, that is only slightly smaller than that of Pt (111).


Sublethal toxicity of esbiothrin relationship with total antioxidant status and in vivo genotoxicity assessment in fish (Cyprinus carpio L., 1758) using the micronucleus test and comet assay.
Esbiothrin, synthetic pyrethroid with quick activity against insects, is widely used against household pests and in public health. Despite widespread use, data on ecotoxicity and genotoxic effects are extremely scarce. The aim of the present study is to evaluate the genotoxic potential of esbiothrin on a model fish species Cyprinus carpio L., 1758 (Pisces: Cyprinidae, koi) using the micronucleus test and comet assay in peripheral blood erythrocytes. Effects of two sublethal exposure concentrations on plasma total antioxidant status (TAS mmol/L), and Hct values were examined. On the basis of the 96 h LC50 data from U.S. EPA ecotox database (32 μg/L) two sublethal exposure concentrations (5 and 10 μg/L) were used together with ethyl methanesulfonate (EMS) (5 mg/L) as positive control. Five fish were used for each dose/duration group (24, 48, and 72 h) under controlled laboratory conditions. The fish showed behavioral changes at the higher dose. Plasma TAS (mmol/L) levels decreased in 24 h; an increase was observed slightly for 48 and obviously for 72 h in both exposure doses. Similarly, hematocrit (Hct) values differed between exposure duration but no significant differences in mean values were found between groups of the same exposure time. The general trend was a rise after 48 h, which decreased afterwards. Our results revealed significant increases in the frequencies of micronuclei and levels of DNA strand breaks and thus demonstrated the genotoxic potential of this pesticide on fish, a nontarget organism of the aquatic ecosystem. To our knowledge this is the first study to report observable genotoxic effects of esbiothrin on fish.


Bricks and Morals-Hospital Buildings, Do No Harm.
The volume and rigor of evidence-based design have increasingly grown over the last three decades since the field's inception, supporting research-based designs to improve patient outcomes. This movement of using evidence from engineering and the hard sciences is not necessarily new, but design-based health research launched with the demonstration that post-operative patients with window views towards nature versus a brick wall yielded shorter lengths of hospital stay and less analgesia use, promoting subsequent investigations and guideline development. Architects continue to base healthcare design decisions on credible research, with a recent shift in physician involvement in the design process by introducing clinicians to design-thinking methodologies. In parallel, architects are becoming familiar with research-based practice, allowing for further rigor and clinical partnership. This cross-pollination of fields could benefit from further discussion surrounding the ethics of hospital architecture as applied to current building codes and guidelines. Historical precedents where the building was used as a form of treatment can inform future concepts of ethical design practice when applied to current population health challenges, such as design for dementia care. While architecture itself does not necessarily provide a cure, good design can act as a preventative tool and enhance overall quality of care.


Association of cannabis use with the development of elevated anxiety symptoms in the general population: a meta-analysis.
The directionality and magnitude of the association of cannabis use with elevated anxiety symptoms in the general population is unclear. The aim of this meta-analysis was to investigate the association of cannabis use with the development of elevated anxiety symptoms in the general population. A 'random effects' meta-analysis of prospective longitudinal studies was undertaken in line with Meta-analysis Of Observational Studies in Epidemiology (MOOSE) guidelines. Six databases were systematically searched up until 20 May 2016: PsycINFO, MEDLINE, EMBASE, CINAHL Plus, Social Science Citation Index and System for Information on Grey Literature in Europe (SIGLE). Searching ceased on 20 May 2016. The exposure was cannabis use (or use frequency), measured at baseline and the outcome was anxiety, using diagnosis or cut-off points on standardised scales measuring symptoms. The main analysis (k=10; N=58 538) demonstrated an association of cannabis use with anxiety, with a very small OR of 1.15 (95% CI 1.03 to 1.29). Restricting the analysis to high-quality studies (k=5) decreased the OR considerably (OR=1.04; 95% CI 0.91 to 1.19), as did adjusting for publication bias (OR=1.08; 95% CI 0.94 to 1.23). Studies with a baseline in the last 10 years yielded a lower pooled OR than studies with an earlier baseline and studies set in the Americas yielded a markedly higher pooled OR than European studies and Australasian studies. The findings indicate that cannabis use is no more than a minor risk factor for the development of elevated anxiety symptoms in the general population. They may inform the debate surrounding the legalisation of cannabis.


Studies on cell communication with enucleated human fibroblasts.
Metabolic cooperation, the correction of the mutant phenotype in cells deficient in hypoxanthine phosphoribosyltransferase (HPRT-) by intimate contact with normal cells (HPRT+), represents a form of cell communication that is easily studied with radioautography. In the present study it was found that the formation of cell junctions needed for communication does not require protein synthesis nor is it under the immediate control of the cell nucleus. Enucleated normal cells efficiently communicate with HPRT- mutant cells. The effectiveness of enucleated cells as donors in metabolic cooperation provides evidence that it is the transfer of small molecules, nucleotide, or nucleotide derivatives that is responsible for correction of the mutant phenotype. Karyoplasts (nuclei with small amounts of cytoplasm surrounded by a plasma membrane) are unable to efficiently communicate with intact cells. The utilization of [3H]hypoxanthine by communicating mixtures of HPRT+ and HPRT- human cells is not significantly different than in the normal cells alone. Metabolic cooperation, as studied involves a redistribution of purine-containing compounds among communicating cells.


Anger: cause or consequence of posttraumatic stress? A prospective study of Dutch soldiers.
Many studies have shown that individuals with posttraumatic stress disorder (PTSD) experience more anger over time and across situations (i.e., trait anger) than trauma-exposed individuals without PTSD. There is a lack of prospective research, however, that considers anger levels before trauma exposure. The aim of this study was to prospectively assess the relationship between trait anger and PTSD symptoms, with several known risk factors, including baseline symptoms, neuroticism, and stressor severity in the model. Participants were 249 Dutch soldiers tested approximately 2 months before and approximately 2 months and 9 months after their deployment to Afghanistan. Trait anger and PTSD symptom severity were measured at all assessments. Structural equation modeling including cross-lagged effects showed that higher trait anger before deployment predicted higher PTSD symptoms 2 months after deployment (β = .36), with stressor severity and baseline symptoms in the model, but not with neuroticism in the model. Trait anger at 2 months postdeployment did not predict PTSD symptom severity at 9 months, and PTSD symptom severity 2 months postdeployment did not predict subsequent trait anger scores. Findings suggest that trait anger may be a pretrauma vulnerability factor for PTSD symptoms, but does not add variance beyond the effect of neuroticism.


School health training during pediatric residency.
Pediatricians are becoming increasingly concerned about reaching beyond the office or clinic to help solve problems of child and family health. The physician's ability to interact with school personnel and communities is one important approach to this outreach effort. School health training has been a required 6-month component of the pediatric residency curriculum at the University of Texas Medical Branch, Galveston, since 1979, with second-year residents providing weekly on-site consultations to public school districts and participating in a weekly school health seminar series. In this study, graduates from this program were surveyed to determine their evaluations of the training and extent of current school involvement and to use their evaluations for curriculum evaluation. Residents who participated in the school consultation from 1979 through 1988 were surveyed. Seventy-eight pediatricians responded, a 79% return rate. Ratings by the graduates reveal that both pediatric generalists and subspecialists highly value their training experiences. Ratings were not related to the specific site to which the resident consultant had been assigned. Of these pediatricians, 41% are currently consulting with school personnel. These findings are discussed as they relate to other reports of physician involvement in schools and provide clues to curriculum design for school health training in pediatric residency.


Health status and quality of life of elderly persons with severe hemophilia born before the advent of modern replacement therapy.
More and more people with severe hemophilia reach an old age thanks to an effective treatment. There is no information on the health status and quality of life of elderly people with hemophilia born at a time when replacement therapy was hardly available. Italian patients with severe hemophilia, aged >or=65 years and hence born in 1942 or earlier, were compared with elderly men without bleeding disorders matched for age, sex, geography and social status. The following aspects were evaluated: concomitant illness, orthopedic status, physical functioning and cognitive status. Measurements of generic and disease-specific health-related quality of life were also obtained, together with the presence or absence of depression. Thirty-nine patients, aged 65-78 years, were investigated; 33 had hemophilia A. All patients had started regular treatment on demand only when they were already 25-30 years of age. Patients were compared with 43 men without hemophilia, aged 65-79 years. More patients with hemophilia had chronic hepatitis B and C, HIV infection and hypertension. On the other hand, their elderly peers without hemophilia were more frequently hypercholesterolemic and had more cardiovascular diseases. Most hemophiliacs had arthropathy and worse values for physical functioning, but their cognitive status was similar to that of elderly non-hemophiliacs. Hemophiliacs reported greater depression and lower health-related quality of life. Elderly patients with hemophilia have more co-morbidities and problems in daily living, but similar cognitive status as age-matched non-hemophilic peers. They have more chronic viral infections and hypertension but fewer cardiovascular diseases. These observations should help to optimize health care delivery in this increasing and neglected population of people with hemophilia.


Ameliorative and protective effects of ginger and its main constituents against natural, chemical and radiation-induced toxicities: A comprehensive review.
Fatal unintentional poisoning is widespread upon human exposure to toxic agents such as pesticides, heavy metals, environmental pollutants, bacterial and fungal toxins or even some medications and cosmetic products. In this regards, the application of the natural dietary agents as antidotes has engrossed a substantial attention. One of the ancient known traditional medicines and spices with an arsenal of metabolites of several reported health benefits is ginger. This extended literature review serves to demonstrate the protective effects and mechanisms of ginger and its phytochemicals against natural, chemical and radiation-induced toxicities. Collected data obtained from the in-vivo and in-vitro experimental studies in this overview detail the designation of the protective effects to ginger's antioxidant, anti-inflammatory, and anti-apoptotic properties. Ginger's armoury of phytochemicals exerted its protective function via different mechanisms and cell signalling pathways, including Nrf2/ARE, MAPK, NF-ƙB, Wnt/β-catenin, TGF-β1/Smad3, and ERK/CREB. The outcomes of this review could encourage further clinical trials of ginger applications in radiotherapy and chemotherapy regime for cancer treatments or its implementation to counteract the chemical toxicity induced by industrial pollutants, alcohol, smoking or administered drugs.


Statistical modeling of interlocus interactions in a complex disease: rejection of the multiplicative model of epistasis in type 1 diabetes.
In general, common diseases do not follow a Mendelian inheritance pattern. To identify disease mechanisms and etiology, their genetic dissection may be assisted by evaluation of linkage in mouse models of human disease. Statistical modeling of multiple-locus linkage data from the nonobese diabetic (NOD) mouse model of type 1 diabetes has previously provided evidence for epistasis between alleles of several Idd (insulin-dependent diabetes) loci. The construction of NOD congenic strains containing selected segments of the diabetes-resistant strain genome allows analysis of the joint effects of alleles of different loci in isolation, without the complication of other segregating Idd loci. In this article, we analyze data from congenic strains carrying two chromosome intervals (a double congenic strain) for two pairs of loci: Idd3 and Idd10 and Idd3 and Idd5. The joint action of both pairs is consistent with models of additivity on either the log odds of the penetrance, or the liability scale, rather than with the previously proposed multiplicative model of epistasis. For Idd3 and Idd5 we would also not reject a model of additivity on the penetrance scale, which might indicate a disease model mediated by more than one pathway leading to beta-cell destruction and development of diabetes. However, there has been confusion between different definitions of interaction or epistasis as used in the biological, statistical, epidemiological, and quantitative and human genetics fields. The degree to which statistical analyses can elucidate underlying biologic mechanisms may be limited and may require prior knowledge of the underlying etiology.


A novel model for Ki67 assessment in breast cancer.
Ki67 is currently the proliferation biomarker of choice, with both prognostic and predictive value in breast cancer. A lack of consensus regarding Ki67 use in pre-analytical, analytical and post-analytical practice may hinder its formal acceptance in the clinical setting. One hundred breast cancer samples were stained for Ki67. A standard estimation of Ki67 using fixed denominators of 200, 400 and 1 000 counted tumor cells was performed, and a cut-off at 20% was applied, Ki67static. A novel stepwise counting strategy for Ki67 estimation, Ki67scs, was developed based on rejection regions derived from exact two-sided binomial confidence intervals for proportions. Ki67scs was defined by the following parameters: the cut-off (20%), minimum (50) and maximum (400) number of tumor cells to count, increment (10) and overall significance level of the test procedure (0.05). Results from Ki67scs were compared to results from the Ki67static estimation with fixed denominators. For Ki67scs, the median number of tumor cells needed to determine Ki67 status was 100; the average, 175. Among 38 highly proliferative samples, the average Ki67scs fraction was 45%. For these samples, the fraction decreased from 39% to 37% to 35% with static counting of 200, 400 and 1 000 cells, respectively. The largest absolute difference between the estimation methods was 23% (42% (Ki67scs) vs. 19% (Ki67static)) and resulted in an altered sample classification. Among the 82 unequivocal samples, 74 samples received the same classification using both Ki67scs and Ki67static. Of the eight disparate samples, seven were classified highly proliferative by Ki67static when 200 cells were counted; whereas all eight cases were classified as low proliferative when 1 000 cells were counted. Ki67 estimation using fixed denominators may be inadequate, particularly for tumors demonstrating extensive heterogeneity. We propose a time saving stepwise counting strategy, which acknowledges small highly proliferative hot spots. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/3588156111195336.


Size distribution of exhaled aerosol particles containing SARS-CoV-2 RNA.
SARS-CoV-2 in exhaled aerosols is considered an important contributor to the spread of COVID-19. However, characterizing the size distribution of virus-containing aerosol particles has been challenging as high concentrations of SARS-CoV-2 in exhaled air is mainly present close to symptom onset. We present a case study of a person with COVID-19 who was able to participate in extensive measurements of exhaled aerosols already on the day of symptom onset and then for the following three days. Aerosol collection was performed using an eight-stage impactor while the subject was breathing, talking and singing, for 30 min each, once every day. In addition, nasopharyngeal samples, saliva samples, room air samples and information on symptom manifestations were collected every day. Samples were analyzed by RT-qPCR for detection of SARS-CoV-2 RNA. SARS-CoV-2 RNA was detected in seven of the eight particle size fractions, from 0.34 to >8.1 µm, with the highest concentrations found in 0.94-2.8 µm particles. The concentration of SARS-CoV-2 RNA was highest on the day of symptom onset, and declined for each day thereafter. Our data showed that 90% of the exhaled SARS-CoV-2 RNA was found in aerosol particles <4.5 µm, indicating the importance of small particles for the transmission of COVID-19 close to symptom onset. These results are important for our understanding of airborne transmission, for developing accurate models and for selecting appropriate mitigation strategies.


Acute Retinopathy in Pseudoxanthoma Elasticum.
Acute retinopathy may partly explain variable disease manifestation and vision loss in patients with pseudoxanthoma elasticum (PXE). The diagnosis of this likely autoimmune process may inform patient counseling and treatment approaches. To characterize acute retinopathy in patients with PXE as a disease manifestation that may be associated with profound visual impairment. This single-center case series was conducted from May 2013 to October 2018. It used the patient database of the Department of Ophthalmology at the University of Bonn, a referral center for PXE in Germany. Patients at this center with genetically confirmed PXE and who met the inclusion criteria were included (n = 9). Patients underwent multimodal retinal imaging, including fundus photography, fundus autofluorescence (AF), optical coherence tomography (OCT), fluorescein angiography (FA), and indocyanine green angiography (ICGA); in select cases, electroretinography as well as antiretinal and anti-retinal pigment epithelium (RPE) antibody testing were also used. Clinical presentation and disease course. Nine patients (8 [89%] female; mean [range] age, 43 [19-55] years) with acute retinopathy were identified in a cohort of 167 consecutive patients with PXE (frequency of 5%). Symptoms ranged from light sensations or metamorphopsia to profound vision loss. Visual acuity was reduced in 6 patients (67%), ranging from a best-corrected visual acuity of 20/30 to perception of hand movements at manifestation. All patients revealed characteristic fundus features with temporary appearance of partly confluent outer retinal whitish dots at the posterior pole, which corresponded to areas of hyperautofluorescence on fundus AF, loss of the ellipsoid band on OCT, and associated scotomata. The FA and late-phase ICGA imaging showed associated hyperfluorescence and hypocyanescence. Electroretinography revealed a variable reduction of amplitudes. Changes were fully reversible within 1 month in 3 of 8 patients with available follow-up data. Of the remaining 5 patients, 3 had a prolonged and likely permanent vision loss (observation period, 1-64 months) mainly owing to central subretinal hyperreflective material originating from angioid streaks. In 4 (67%) of 6 tested, antiretinal and/or anti-RPE antibodies were detected. Acute retinopathy in patients with PXE may occur, with symptoms ranging from short-term, reversible alterations to irreversible vision loss; these findings contribute to understanding the variable ocular disease progression in PXE and provide insights into the autoimmune phenomena of the posterior pole.


"She would sit with me": mothers' experiences of individual peer support for exclusive breastfeeding in Uganda.
Different strategies have been used to improve the initiation and duration of breastfeeding. Peer counsellors are reported to improve exclusive breastfeeding levels, but few studies have assessed the satisfaction of women with the support given, especially in Africa. In this paper we describe women's experiences of peer counselling for exclusive breastfeeding in an East African setting. In the Ugandan site of PROMISE-EBF, a multi-centre community randomised trial to evaluate the effect of peer counselling for exclusive breastfeeding on infant health, 370 women in the intervention arm participated in a study exit interview. Individual peer counselling was offered to women in 12 of the 24 study clusters, scheduled as five visits: before childbirth and during weeks 1, 4, 7 and 10 after childbirth. During the visits, the women were given information and skills to help them breastfeed exclusively. After the 10-week visit, they were interviewed about their feelings and experiences related to the peer counselling. Overall, more than 95% of the women expressed satisfaction with the various aspects of peer counselling offered. Those who had received five or more visits were more likely to give positive responses about their experience with peer counselling than those who had received fewer visits. They explained their satisfaction with time spent with the peer counsellor in terms of how much she discussed with them. Most women felt their knowledge needs about breastfeeding were covered by the peer counsellors, while others expressed a desire to learn about complementary feeding and family planning. Attributes of the peer counsellors included their friendliness, being women and giving support in a familiar and relaxed way. Women were positive about the acquisition of knowledge and the benefit to their babies from the peer counselling. They preferred a peer counsellor to a health worker for support of exclusive breastfeeding because of their friendly approach. Individual peer counselling to support exclusive breastfeeding was positively received by the women. clinicaltrials.gov no: NCT00397150.


Challenging future, challenging past: the relationship of social integration and psychological impairment in traumatized refugees.
Refugees have been shown to present high prevalence rates of trauma-related mental disorders. Despite their psychological impairment, they are expected to meet high functional requirements in terms of social integration into, and financial independence from, the host society. This cross-sectional study examined the relationship of mental health problems, post-migration living difficulties (PMLD), and social integration in a sample of 104 refugees seeking treatment for severe posttraumatic stress and comorbid symptoms in two outpatient clinics in Switzerland. Despite an average time of residence in Switzerland of over 10 years, participants showed poor integration and a high number of PMLD. Integration difficulties were closely associated with psychological symptoms, but not with socio-demographic parameters such as education or visa status. Psychological impairment in treatment-seeking traumatized refugees is associated with poor integration. To foster social integration, it is crucial to better understand and address the specific needs of this highly vulnerable population.


[Parinaud syndrome caused by corpus pineale tumour in a 14-year-old boy].
Parinaud syndrome (PS) can manifest in a pineal tumour. Major components of PS include restriction of gazing upwards, light-near dissociation and convergence retraction nystagmus. A 14-year-old boy presented with diplopia and restricted ability to gaze upwards. The objective examination revealed signs, which were compatible with the major manifestations of PS. A magnetic resonance scan (MRI) of cerebrum indicated pinealoma, and a pathological examination identified the lesion as a germ cell carcinoma. The patient received chemotherapy and stereotactic radiosurgery. The ophthalmic symptoms improved, and a follow-up MRI demonstrated complete regression of the tumour.


A New Risk Score to Predict In-Hospital Mortality in Elderly Patients With Acute Heart Failure: On Behalf of the Journey HF-TR Study Investigators.
We aimed to predict in-hospital mortality of elderly patients with heart failure (HF) by using a risk score model which could be easily applied in routine clinical practice without using an electronic calculator. The study population (n = 1034) recruited from the Journey HF-TR (Patient Journey in Hospital with Heart Failure in Turkish Population) study was divided into a derivation and a validation cohort. The parameters related to in-hospital mortality were first analyzed by univariate analysis, then the variables found to be significant in that analysis were entered into a stepwise multivariate logistic regression (LR) analysis. Patients were classified as low, intermediate, and high risk. A risk score obtained by taking into account the regression coefficients of the significant variables as a result of the LR analysis was tested in the validation cohort using receiver operating characteristic curve analysis. In total, 6 independent variables (age, blood urea nitrogen, previous history of hemodialysis/hemofiltration, inotropic agent use, and length of intensive care stay) associated with in-hospital mortality were included in the analysis. The risk score had a good discrimination in both the derivation and validation cohorts. A new validated risk score to determine the risk of in-hospital mortality of elderly hospitalized patients with HF was developed by including 6 independent predictors.


Minimal influence of water and nutrient content on the bacterial community composition of a maritime Antarctic soil.
Bacterial community composition was determined by culture-independent PCR-based methods in two soils differing markedly in their water, C, N and P contents sampled from Mars Oasis on Alexander Island, western Antarctic Peninsula. 16S rRNA sequences of the phyla Actinobacteria, Cyanobacteria, α-Proteobacteria and Acidobacteria were commonly (> 8% frequency) obtained from soil. Those of β-, γ- and δ-Proteobacteria, Chloroflexi, Bacteroidetes, Verrucomicrobia, Planctomycetes, Gemmatimonadetes and Firmicutes were less frequent. Comparisons of slopes of collector's curves and the Shannon-Weiner diversity index indicated no difference in overall bacterial diversity between the two soils, although sequences of δ-Proteobacteria and the cyanobacterial genus Leptolyngbya were more commonly derived from the soil with the higher water and nutrient content. The data suggest that different levels of soil water, C, N and P have only a minor effect on the bacterial community composition of maritime Antarctic soils.


Epigenetic mechanisms and metabolic reprogramming in fibrogenesis: dual targeting of G9a and DNMT1 for the inhibition of liver fibrosis.
Hepatic stellate cells (HSC) transdifferentiation into myofibroblasts is central to fibrogenesis. Epigenetic mechanisms, including histone and DNA methylation, play a key role in this process. Concerted action between histone and DNA-mehyltransferases like G9a and DNMT1 is a common theme in gene expression regulation. We aimed to study the efficacy of CM272, a first-in-class dual and reversible G9a/DNMT1 inhibitor, in halting fibrogenesis. G9a and DNMT1 were analysed in cirrhotic human livers, mouse models of liver fibrosis and cultured mouse HSC. G9a and DNMT1 expression was knocked down or inhibited with CM272 in human HSC (hHSC), and transcriptomic responses to transforming growth factor-β1 (TGFβ1) were examined. Glycolytic metabolism and mitochondrial function were analysed with Seahorse-XF technology. Gene expression regulation was analysed by chromatin immunoprecipitation and methylation-specific PCR. Antifibrogenic activity and safety of CM272 were studied in mouse chronic CCl4 administration and bile duct ligation (BDL), and in human precision-cut liver slices (PCLSs) in a new bioreactor technology. G9a and DNMT1 were detected in stromal cells in areas of active fibrosis in human and mouse livers. G9a and DNMT1 expression was induced during mouse HSC activation, and TGFβ1 triggered their chromatin recruitment in hHSC. G9a/DNMT1 knockdown and CM272 inhibited TGFβ1 fibrogenic responses in hHSC. TGFβ1-mediated profibrogenic metabolic reprogramming was abrogated by CM272, which restored gluconeogenic gene expression and mitochondrial function through on-target epigenetic effects. CM272 inhibited fibrogenesis in mice and PCLSs without toxicity. Dual G9a/DNMT1 inhibition by compounds like CM272 may be a novel therapeutic strategy for treating liver fibrosis.


Concurrent micronutrient deficiencies are prevalent in nonpregnant rural and tribal women from central India.
The existence of concurrent micronutrient deficiencies in Indian women of reproductive age has received little attention. This study aimed to comprehensively assess the micronutrient status of nonpregnant rural and tribal women 18-30 y from central India. Participants (n = 109) were randomly selected using a stratified (rural-tribal) proportionate-to-population size cluster sampling method from 12 subcenters in Ramtek block, Nagpur. Sociodemographic, anthropometric, dietary, and biochemical data, including blood and urine samples, were obtained. Tribal and rural women had similar sociodemographic characteristics and anthropometric status; 63% of women had a body mass index <18.5 kg/m(2). The median urinary iodine concentration was 215 μg/L (IQR: 127, 319). The mean (SD) concentration of hemoglobin, serum zinc, retinol, and folate was 112 (13) g/L, 10.8 (1.6) μmol/L, 1.2 (0.3) μmol/L, 18.4 (8.4) nmol/L, respectively, with a geometric mean serum vitamin B(12) concentration of 186 pmol/L. The percentage of women with low values for hemoglobin (<120 g/L), serum zinc (<10.7 μmol/L), vitamin B(12) (<148 pmol/L), retinol (<0.7 μmol/L), and folate (<6.8 nmol/L) was 66%, 52%, 34%, 4%, and 2%, respectively. Tribal women had a higher prevalence of zinc deficiency (58% versus 39%, P = 0.054) and concurrent deficiency of any two micronutrients (46% versus 26%; P = 0.034), including zinc and anemia (38% versus 21%, P = 0.024). Zinc, vitamin B(12), and iron constitute the principal micronutrient deficiencies in these women. Existing supplementation programs should be extended to include 18- to 30-y-old nonpregnant women as the majority of childbearing occurs within this timeframe.


TA-periodic ("601"-like) centromeric nucleosomes of A. thaliana.
The bulk of strong nucleosomes (SNs, with visibly periodic DNA sequences) is described by consensus pattern of 5 or 6 base runs of purines alternating with similar runs of pyrimidines - RR/YY SNs. Yet, the strongest known nucleosome positioning sequence, the 601 clone of Lowary and Widom, is rather periodic repetition of TA dinucleotides following one another every 10 bases. We located "601"-like TA-periodic sequences in the genome of A. thaliana. Several families of such sequences are discovered repeating almost exclusively in centromeres. Thus, while A. thaliana SNs of RR/YY type have strong affinity to pericentromeric regions, as it has been previously found, the SNs of TA periodic type concentrate rather in centromeres.


Variations in mortality and length of stay in intensive care units.
To evaluate the amount of variation in in-hospital mortality and length of intensive care unit (ICU) stay that can be accounted for by clinical data available at ICU admission. Inception cohort study. Forty-two ICUs in 40 hospitals, including 26 hospitals that were randomly selected and 14 large tertiary care hospitals that volunteered for the study. A consecutive sample of 16,622 patients and 17,440 ICU admissions. Data on selected demographic characteristics, comorbidity, and specific physiologic variables were recorded during the first ICU day for an average of 415 admissions at each ICU; hospital discharge status (dead or alive) and length of ICU stay were recorded for individual patients; and the ratio of actual to predicted in-hospital mortality, standardized mortality ratios, and the ratio of actual to predicted length of ICU stay were recorded for individual ICUs. Unadjusted in-hospital mortality rates for the 42 units varied from 6.4% to 40%, and 90% (R2 = 0.90) of this variation was attributable to patient characteristics at admission. The standard mortality ratio varied from 0.67 to 1.25. The mean unadjusted length of ICU stay varied from 3.3 to 7.3 days, and 78% of the variation (R2 = 0.78) was attributed to patient and selected institutional characteristics. The best performing unit had a length of stay ratio of 0.88, whereas the poorest performing unit had a ratio of 1.21. Clinicians can use readily available admission data to adjust for considerable variations in patient severity and type in different ICUs. Such data should permit precise evaluation and comparison of ICU effectiveness and efficiency, which varied substantially in this study, and result in improved methods of risk prediction and evaluation of new medical practices.


Synergistic complement lysis by monoclonal antibodies to the human leukocyte common antigen requires both the classical and alternative pathways.
In a previous study we isolated a series of rat monoclonal antibodies to the human leukocyte common (LC) antigen and demonstrated that synergistic complement lysis was possible between IgG2b antibodies which recognised different epitopes. In this report we have examined the mechanisms that were involved in synergistic lysis. We found that the number of C1q binding sites increased from 30,000 to 40,000/cell using a single antibody to 90,000/cell when two IgG2b antibodies to different epitopes were used together. The affinity of C1q binding also increased approx. 3-fold for the synergistic pair, and there was a similar increase in the rate of C1 activation. Combinations of an IgG2b with IgG1 or IgG2a gave much smaller increases in the amount of C1q bound and in the rate of C1 activation. Despite the large number of C1q molecules bound with the optimal synergistic pair and the increased rate of C1 activation, lysis was inefficient in serum depleted of Factor D, suggesting a requirement for the alternative pathway. This is the first demonstration of the need for the alternative pathway in complement lysis by monoclonal antibodies.


Evidence of the Zanclean megaflood in the eastern Mediterranean Basin.
The Messinian salinity crisis (MSC) - the most abrupt, global-scale environmental change since the end of the Cretaceous - is widely associated with partial desiccation of the Mediterranean Sea. A major open question is the way normal marine conditions were abruptly restored at the end of the MSC. Here we use geological and geophysical data to identify an extensive, buried and chaotic sedimentary body deposited in the western Ionian Basin after the massive Messinian salts and before the Plio-Quaternary open-marine sedimentary sequence. We show that this body is consistent with the passage of a megaflood from the western to the eastern Mediterranean Sea via a south-eastern Sicilian gateway. Our findings provide evidence for a large amplitude drawdown in the Ionian Basin during the MSC, support the scenario of a Mediterranean-wide catastrophic flood at the end of the MSC, and suggest that the identified sedimentary body is the largest known megaflood deposit on Earth.


A pulsed electron gun for ultrafast electron diffraction at surfaces.
The construction of a pulsed electron gun for ultrafast reflection high-energy electron diffraction experiments at surfaces is reported. Special emphasis is placed on the characterization of the electron source: a photocathode, consisting of a 10 nm thin Au film deposited onto a sapphire substrate. Electron pulses are generated by the illumination of the film with ultraviolet laser pulses of femtosecond duration. The photoelectrons are emitted homogeneously across the photocathode with an energy distribution of 0.1 eV width. After leaving the Au film, the electrons are accelerated to kinetic energies of up to 15 keV. Focusing is accomplished by an electrostatic lens. The temporal resolution of the experiment is determined by the probing time of the electrons traveling across the surface which is about 30 ps. However, the duration of the electron pulses can be reduced to less than 6 ps.


Orthotopic liver transplantation for primary sclerosing cholangitis. A 12-year single center experience.
The purpose of this study was to analyze a single center's 12-year experience with 127 orthotopic liver transplantations (OLT) for primary sclerosing cholangitis (PSC). Primary sclerosing cholangitis is a chronic cholestatic liver disease of unknown origin that occurs most commonly in young men and is associated frequently (70-80%) with inflammatory bowel disease (IBD). Patients with PSC also are at risk for the development of cholangiocarcinoma (CCA) and those with IBD for colon carcinoma. Although the course of PSC is variable, it frequently is progressive, leading to cirrhosis and requirement for OLT. The medical records of 127 consecutive patients undergoing OLT for PSC from July 1, 1984, to May 30, 1996, were reviewed. Actuarial patient and graft survival was determined at 1,2, and 5 years. The incidence and outcome of patients with CCA, recurrent sclerosing cholangitis, and post-transplant colon carcinoma was determined. Results were analyzed by way of stepwise Cox regression to determine the statistical strength of independent associations between pretransplant covariates and patient survival. The median follow-up period was 3.01 years. Incidental cholangiocarcinoma (ICCA) was defined as a tumor < 1 cm in size that was discovered at the time of pathologic sectioning of the explanted liver. Ninety-two patients (72%) had associated IBD. Seventy-nine (62%) had undergone previous biliary tract surgery. One hundred seven patients (84%) received a single graft, whereas 20 patients (16%) required 22 retransplants. Patients received either cyclosporine- (n = 76) or tacrolimus- (n = 51) based immunosuppression. The 1-, 2-, and 5-year actuarial patient survivals were 90%, 86%, and 85%, respectively, whereas graft survival was 82%, 77%, and 72%, respectively. The presence of previous biliary surgery had no effect on patient survival. Ten patients (8%) had ICCA and their survival was not significantly different from patients without ICCA (100%, 83%, and 83% at 1, 2, and 5 years, respectively). Four patients were known to have CCA at the time of OLT, all recurred within 6 months, and had a significantly worse outcome (p < 0.0001). Recurrent sclerosing cholangitis developed in 11 patients (8.6%). The patient and graft survival in this group was not different from those in whom recurrence did not develop (patient; 100%, 90%, and 90%; graft: 80%, 70%, and 52%). Thirty patients (23%) underwent colectomy after liver transplantation for dysplasia-carcinoma or symptomatic colitis. Of the nine covariates entered into the Cox multivariate regression analysis, only common bile duct frozen section biopsy specimen showing CCA was predictive of a survival disadvantage. Liver transplantation provides excellent patient and graft survival rates for patients affected with PSC independent of pretransplant biliary tract surgery. Incidental cholangiocarcinoma does not affect patient survival significantly. However, known CCA or common duct frozen section biopsy specimen or both showing CCA are associated with poor recipient survival, and OLT should be proscribed in these cases. Recurrent PSC occurs in approximately 9% of cases but does not affect patient survival. Post-transplant colectomy does not affect patient survival adversely.


Smoking cessation in inpatient psychiatry treatment facilities: A review.
Tobacco-related diseases are a leading cause of death among individuals with severe mental illness (SMI), yet interventions to address tobacco cessation are rare in inpatient settings where persons with SMI are hospitalized. While cigarette smoking rates have declined in the general population, they remain high in persons with SMI. Inpatient settings would be a prime location to intervene on tobacco consumption among persons with SMI. The objective of this review was to examine evidence of smoking cessation interventions in psychiatric inpatient facilities. Using narrative overview guidelines, we searched PubMed, PsycINFO, and CINAHL for smoking cessation RCT studies published between 1950 and 2018. Studies included had to have at least started in inpatient psychiatry settings. Examples of search terms included: smoking cessation in inpatient psychiatry, smoking cessation in inpatient mental health treatment facilities, and smoking cessation and mental health. Following the inclusion criteria, eight RCT studies were reviewed. One study was among adolescent psychiatric inpatient smokers ages 13-17, and 7 were among adult psychiatric inpatients with mean age 41 years. Treatment periods lasting 8 to 12 weeks started in inpatient settings and continued post discharge. A combination of behavioral and pharmacological interventions were used. Pharmacological interventions were nicotine replacement therapies, and at least one study used varenicline. At baseline, participants smoked an average of 18.1 cigarettes per day. Smoking cessation in inpatient psychiatry settings is rare or delayed. There is a need for more tailored treatments among this population to help them quit smoking.


Android and ODK based data collection framework to aid in epidemiological analysis.
Periodic collection of field data, analysis and interpretation of data are key to a good healthcare service. This data is used by the subsequent decision makers to recognize preventive measures, provide timely support to the affected and to help measure the effects of their interventions. While the resources required for good disease surveillance and proactive healthcare are available more readily in developed countries, the lack of these in developing countries may compromise the quality of service provided. This combined with the critical nature of some diseases makes this an essential issue to be addressed. Taking advantage of the rapid growth of cell phone usage and related infrastructure in developed as well as developing countries, several systems have been established to address the gaps in data collection. Android, being an open sourced platform, has gained considerable popularity in this aspect. Open data kit is one such tool developed to aid in data collection. The aim of this paper is to present a prototype framework built using few such existing tools and technologies to address data collection for seasonal influenza, commonly referred to as the flu.


Spinal fractures in patients with diffuse idiopathic skeletal hyperostosis: Clinical characteristics by fracture level.
Diffuse idiopathic skeletal hyperostosis (DISH) makes the spine prone to unstable fractures with neurological deterioration. This study was conducted to assess clinical and radiographic features of spinal fractures in DISH by the level of spinal injury, and to evaluate the optimal treatment for each level. A multicenter retrospective study over a 5-year period, including 46 patients (35 males; 11 females) with a mean age of 77.2 ± 9.7 years at the time of injury. By fracture level, there were 7 cervical (15.2%), 25 thoracic (54.3%), and 14 lumbar (30.4%) fractures. We recorded the cause of injury, whether diagnosis was delayed, and neurological status by Frankel grade. Ossification and fracture patterns were assessed by CT-multi-planar reconstruction (MPR). Neurological status immediately after the cervical-spine injury was C (28.6%) or E (71.4%); after thoracic injury, C (12.0%) or E (88.0%); and after lumbar injury, D (21.4%) or E (78.6%). Inability to walk at admission was more frequent in patients with a spinal-cord injury above the lumbar level (P = .033). Vertebral-body fractures were observed in 14.3% of the cervical injuries, 80.0% of the thoracic injuries, and 50.0% of the lumbar injuries (P = .004). Most patients with a cervical fracture had a disc-level fracture (85.7%). Posterior-column ankylosis was observed in 14.3% of the cervical-fracture group, 72.0% of the thoracic-fracture group, and 78.6% of the lumbar-fracture group (P = .008). Ossification and fracture patterns in patients with DISH varied distinctly by the level of spinal injury. Intervertebral-disc fractures were frequently observed in the cervical spine. Delayed diagnosis, vertebral-body fracture, and posterior-column ankylosis were observed in the thoracolumbar spine. This study recommends 3 above and 3 below fusion, to avoid instrumentation failure in the fixation of spinal fracture in patients with DISH.


Registry-Based Prospective, Active Surveillance of Medical-Device Safety.
The process of assuring the safety of medical devices is constrained by reliance on voluntary reporting of adverse events. We evaluated a strategy of prospective, active surveillance of a national clinical registry to monitor the safety of an implantable vascular-closure device that had a suspected association with increased adverse events after percutaneous coronary intervention (PCI). We used an integrated clinical-data surveillance system to conduct a prospective, propensity-matched analysis of the safety of the Mynx vascular-closure device, as compared with alternative approved vascular-closure devices, with data from the CathPCI Registry of the National Cardiovascular Data Registry. The primary outcome was any vascular complication, which was a composite of access-site bleeding, access-site hematoma, retroperitoneal bleeding, or any vascular complication requiring intervention. Secondary safety end points were access-site bleeding requiring treatment and postprocedural blood transfusion. We analyzed data from 73,124 patients who had received Mynx devices after PCI procedures with femoral access from January 1, 2011, to September 30, 2013. The Mynx device was associated with a significantly greater risk of any vascular complication than were alternative vascular-closure devices (absolute risk, 1.2% vs. 0.8%; relative risk, 1.59; 95% confidence interval [CI], 1.42 to 1.78; P<0.001); there was also a significantly greater risk of access-site bleeding (absolute risk, 0.4% vs. 0.3%; relative risk, 1.34; 95% CI, 1.10 to 1.62; P=0.001) and transfusion (absolute risk, 1.8% vs. 1.5%; relative risk, 1.23; 95% CI, 1.13 to 1.34; P<0.001). The initial alerts occurred within the first 12 months of monitoring. Relative risks were greater in three prespecified high-risk subgroups: patients with diabetes, those 70 years of age or older, and women. All safety alerts were confirmed in an independent sample of 48,992 patients from April 1, 2014, to September 30, 2015. A strategy of prospective, active surveillance of a clinical registry rapidly identified potential safety signals among recipients of an implantable vascular-closure device, with initial alerts occurring within the first 12 months of monitoring. (Funded by the Food and Drug Administration and others.).


Poor outcome of hormone receptor-positive breast cancer at very young age is due to tamoxifen resistance: nationwide survival data in Korea--a report from the Korean Breast Cancer Society.
Breast cancer in very young women (age < 35 years) is uncommon and poorly understood. We sought to evaluate the prognosis and treatment response of these patients compared with women ages 35 to 50 years. We analyzed data from 9,885 breast cancer patients age < or = 50 years who were part of the Korean Breast Cancer Society registration program between 1992 and 2001. The overall survival (OS) and breast cancer-specific survival (BCSS) were compared between age groups. One thousand four hundred forty-four patients (14.6%) were younger than age 35 and 8,441 (85.4%) patients were between 35 and 50 years of age. Younger patients had significantly higher T-stage and higher lymph node positivity and lower hormone receptor expression than older patients. Younger patients had a greater probability of death than older patients, regardless of tumor size or lymph node status. The survival difference was significant for patients with positive or unknown hormone receptor status (P < .0001), but not for patients with negative hormone receptor status. In a multivariate analysis, the interaction term of young age and hormone receptor positivity was significant for OS and BCSS with a hazard ratio for OS of 2.13 (95% CI, 1.52 to 2.98). The significant survival benefit from adjuvant hormone therapy after chemotherapy observed in older patients (hazard ratio for OS, 0.61; 95% CI, 0.47 to 0.79; P = .001) could not be seen in younger patients (P > .05). Younger patients (age < 35) showed worse prognosis than older patients (age, 35 to 50 years) only in the hormone receptor-unknown or hormone receptor-positive subgroups. Adjuvant tamoxifen therapy might provide less survival benefit when added to chemotherapy in very young breast cancer patients.


Resource use and guideline concordance in evaluation of pulmonary nodules for cancer: too much and too little care.
Pulmonary nodules are common, and more will be found with implementation of lung cancer screening. How potentially malignant pulmonary nodules are evaluated may affect patient outcomes, health care costs, and effectiveness of lung cancer screening programs. Guidelines for evaluating pulmonary nodules for cancer exist, but little is known about how nodules are evaluated in the usual care setting. To characterize nodule evaluation and concordance with guidelines. A retrospective cohort study was conducted including detailed review of medical records from pulmonary nodule detection through evaluation completion, cancer diagnosis, or study end (December 31, 2012). The participants included 300 adults with pulmonary nodules from 15 Veterans Affairs hospitals. Resources used for evaluation at any Veterans Affairs facility and guideline-concordant evaluation served as the main outcomes. Twenty-seven of 300 patients (9.0%) with pulmonary nodules ultimately received a diagnosis of lung cancer: 1 of 57 (1.8%) with a nodule of 4 mm or less, 4 of 134 (3.0%) with a nodule of 5 to 8 mm, and 22 of 109 (20.2%) with a nodule larger than 8 mm. Nodule evaluation entailed 1044 imaging studies, 147 consultations, 76 biopsies, 13 resections, and 21 hospitalizations. Radiographic surveillance (n = 277) lasted a median of 13 months but ranged from less than 0.5 months to 8.5 years. Forty-six patients underwent invasive procedures (range per patient, 1-4): 41.3% (19 patients) did not have cancer and 17.4% (8) experienced complications, including 1 death. Notably, 15 of the 300 (5.0%) received no purposeful evaluation and had no obvious reason for deferral, seemingly "falling through the cracks." Among 197 patients with a nodule detected after release of the Fleischner Society guidelines, 44.7% received care inconsistent with guidelines (17.8% overevaluation, 26.9% underevaluation). In multivariable analyses, the strongest predictor of guideline-inconsistent care was inappropriate radiologist recommendations (overevaluation relative risk, 4.6 [95% CI, 2.3-9.2]; underevaluation, 4.3 [2.7-6.8]). Other systems factors associated with underevaluation included receiving care at more than 1 facility (2.0 [1.5-2.7]) and nodule detection during an inpatient or preoperative visit (1.6 [1.1-2.5]). Pulmonary nodule evaluation is often inconsistent with guidelines, including cases with no workup and others with prolonged surveillance or unneeded procedures that may cause harm. Systems to improve quality (eg, aligning radiologist recommendations with guidelines and facilitating communication across providers) are needed before lung cancer screening is widely implemented.