Life cycle assessment part 2: current impact assessment practice.
Providing our society with goods and services contributes to a wide range of environmental impacts. Waste generation, emissions and the consumption of resources occur at many stages in a product's life cycle-from raw material extraction, energy acquisition, production and manufacturing, use, reuse, recycling, through to ultimate disposal. These all contribute to impacts such as climate change, stratospheric ozone depletion, photooxidant formation (smog), eutrophication, acidification, toxicological stress on human health and ecosystems, the depletion of resources and noise-among others. The need exists to address these product-related contributions more holistically and in an integrated manner, providing complimentary insights to those of regulatory/process-oriented methodologies. A previous article (Part 1, Rebitzer et al., 2004) outlined how to define and model a product's life cycle in current practice, as well as the methods and tools that are available for compiling the associated waste, emissions and resource consumption data into a life cycle inventory. This article highlights how practitioners and researchers from many domains have come together to provide indicators for the different impacts attributable to products in the life cycle impact assessment (LCIA) phase of life cycle assessment (LCA).


The bacteriophage lambda O and P protein initiators promote the replication of single-stranded DNA.
A soluble enzyme system that specifically initiates lambda dv plasmid DNA replication at a bacteriophage lambda replication origin [Wold et al. (1982) Proc. Natl. Acad. Sci. USA 79, 6176-6180] is also capable of replicating the single-stranded circular chromosomes of phages M13 and phi X174 to a duplex form. This chain initiation on single-stranded templates is novel in that it is absolutely dependent on the lambda O and P protein chromosomal initiators and on several Escherichia coli proteins that are known to function in the replication of the lambda chromosome in vivo, including the host dnaB, dnaG (primase), dnaJ and dnaK replication proteins. Strand initiation occurs at multiple sites following an O and P protein-dependent pre-priming step in which the DNA is converted into an activated nucleoprotein complex containing the bacterial dnaB protein. We propose a scheme for the initiation of DNA synthesis on single-stranded templates in this enzyme system that may be relevant to strand initiation events that occur during replication of phage lambda in vivo.


Increased intra-articular granzyme M may trigger local IFN-λ1/IL-29 response in rheumatoid arthritis.
Granzymes are serine proteases involved in eliminating tumour cells and virally infected cells. In addition, extracellular granzyme levels are elevated in inflammatory conditions, including several types of infection and autoimmune diseases, such as rheumatoid arthritis (RA). While GrA and GrB have been associated with RA, a role for the other three granzymes (GrH, GrK, and GrM) in this disease remains unclear. Here, we aimed to investigate the presence and role of GrM and GrK in serum and synovial fluid of patients with RA, psoriatic arthritis, and osteoarthritis. Granzyme levels were determined in serum, synovial fluid, peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) of RA patients and relevant control groups. In addition, the link between GrM and inflammatory cytokines in synovial fluid was investigated. Serum GrM and GrK levels were not affected in RA. GrM, but not GrK, levels were elevated in synovial fluid of RA patients. GrM was mainly expressed by cytotoxic lymphocytes in SFMCs with a similar expression pattern as compared with PBMCs. Intra-articular GrM expression correlated with IL-25, IL-29, XCL1, and TNFα levels. Intriguingly, purified GrM triggered the release of IL-29 (IFN-λ1) from human fibroblasts in vitro. These data indicate that GrM levels are increased in RA synovial fluid and that GrM can stimulate proinflammatory IL-29 release from fibroblasts, suggesting a role of GrM in the pathogenesis of RA.


Puerarin prevents cadmium-induced disorder of testicular lactic acid metabolism in rats by activating 5' AMP-activated protein kinase (AMPK)/sirtuin 1 (SIRT1) signaling pathway.
Cadmium (Cd) interferes with the function of the male reproductive system; however, the molecular mechanism is poorly understood. This study aimed to evaluate the effect of puerarin (PU) on Cd-induced testicular lactic acid metabolism disorder. Weaning male Sprague-Dawley rats were pre-fed for 7 days, weighed, and randomly divided into four groups: Control group, CdAc2 group, CdAc2 + PU group, PU group. The results showed that Cd accumulated in the testis, the testicles became congested and shrank, and the testis index decreased in the rats treated in the CdAc2 group. Cadmium exposure reduced the serum concentration of testosterone, and the concentration of lactic acid and pyruvate in the testis. Cd decreased testicular superoxide dismutase activity and total antioxidant capacity, and increased testicular malondialdehyde levels. Cd reduced the level of ATP, glycolytic gene expression, and lactate production-related proteins in the testis. Cd also decreased the expression of 5' AMP-activated protein kinase (AMPK)/sirtuin 1 (SIRT1) signaling pathway-related proteins in the testis. However, these negative effects were attenuated by PU administration. In summary, Cd reduces the production of lactic acid in the testis of rats, while PU administration restores the production of lactic acid and reduces the toxicity of Cd to the testis of rats.


Investigation of comparative effectiveness research in Asia, Europe, and North America.
Comparative effectiveness research (CER) is an important branch of pharmacoeconomics that systematically studies and evaluates the cost-effectiveness of medical interventions. CER plays instrumental roles in guiding government public health policy programs and insurance. Countries throughout the world use different methods of CER to help make medical decisions based on providing optimal therapy at a reduced cost. Expenses to the healthcare system continue to rise, and CER is one-way in which expenses could be curbed in the future by applying cost-effectiveness evidence to clinical decisions. China, India, South Korea, and the United Kingdom are of essential focus because these country's economies and health care expenses continue to expand. The structures and use of CER are diverse throughout these countries, and each is of prime importance. By conducting this thorough comparison of CER in different nations, strategies and organizational setups from different countries can be applied to help guide public health and medical decision-making in order to continue to expand the establishment and role of CER programs. The patient-centered medical home has been created to help reduce costs in the primary care sector and to help improve the effectiveness of therapy. Barriers to CER are also important as many stakeholders need to be able to work together to provide the best CER evidence. The advancement of CER in multiple countries throughout the world provides a possible way of reducing costs to the healthcare system in an age of expanding expenses.


Hatching system and time effects on broiler physiology and posthatch growth.
A multilevel housing system for broilers was developed, named Patio (Vencomatic BV, Eersel, the Netherlands), in which the hatching and brooding phase are combined. In a Patio system, climate conditions differ from those provided in the hatchers currently in use. We compared the physiology of broilers hatched in a hatcher or in a Patio system, and included the effects of hatching time. Eggs from 1 breeder flock were incubated until embryonic d 18 in a setter and subsequently placed in a hatcher or the Patio until the end of incubation. From each hatching system, 154 chicks were collected per hatching time, at 465 h (early), 480 h (midterm), and 493 h (late) of incubation, from which 24 chicks/group were decapitated for analyses of blood plasma and organ weights. The remaining 130 chicks in each group from both systems were individually labeled and placed together in the Patio system. All chicks were given access to feed and water directly after hatch and were housed up to d 45 to monitor growth. From embryonic d 18 until the end of incubation, average ambient temperature and RH were 38.1°C and 50.8% in the hatcher and 35.2°C and 29.7% in the Patio system. Glucose and corticosterone were slightly higher in hatcher chicks, whereas organ weights were not affected by the hatching system. Although hatchling weights were lower in hatchery chicks, growth from d 0 to 45 was not affected by the hatching system. In both systems, glucose increased with hatching time, whereas lactate and triiodothyronine levels decreased. Yolk weights decreased with hatching time, whereas absolute and relative weights of the yolk-free body, intestines, stomach, lungs, and heart increased, indicating more advanced maturation of organs. Growth up to d 21 was depressed in chicks in the late group, which was possibly related to lower thyroid hormone levels at hatching. We conclude that the hatching system had minor effects on hatchling physiology and that posthatch growth and livability were not affected. Because hatching time affected broiler physiology, it seems important to take hatching time into account in future studies related to hatchling physiology.


Blood gas analysis: a study of blood loss in intensive care.
This paper describes a quantitative study conducted on an intensive care unit in the north of England. It involved the collection of data from the existing records of 65 patients consecutively sampled from a predetermined date provided that they stayed more than 24 hours and had an arterial line in situ. As patient records were used, ethical approval was not necessary. The objectives of the study were to quantify the mean number of blood gas samples taken per patient and estimate the mean blood loss resulting from this, including discard volume. Limitations include reliance on records and lack of an economic evaluation. The results show that blood loss in this study was greater than that reported elsewhere. Patients who were ventilated for 24 hours or more had a statistically significant greater blood loss when compared to those who were not (P < 0.001). A subgroup of patients undergoing renal replacement therapy had the greatest blood loss (mean 55.18 ml per day). This loss was statistically significant when compared to patients not in acute renal failure (P=0.007). When patients undergoing multiple therapies normally associated with increased sampling were compared to patients not receiving such therapies, there was no statistically significant difference in blood loss. The need to change current nursing practice to reduce iatrogenic anaemia is emphasized.


CTLA4 exon 1 dimorphism in bullous and cicatricial pemphigoid.
The human cytotoxic T-lymphocyte antigen 4 (CTLA4) gene encodes proteins regulating the immune response. The polymorphism of this gene is associated with some autoimmune diseases. In this study, we analysed the distribution of the dimorphisms of exon 1 (+ 49 A/G) in bullous pemphigoid (BP) and cicatricial pemphigoid (CP), two types of autoimmune bullous skin diseases that occur in elderly people. The frequency of the exon 1 A-G genotype was marginally decreased in patients (36.4%; n = 55) compared with controls (52.8%, n = 53), but the results were not statistically significant (P = 0.09).


Emotional reactions of Arab lay persons to a person with Alzheimer's disease.
To assess emotional reactions of Israeli Arab people to persons with Alzheimer's Disease (AD) and assess the associations of perceptions of threat of AD, personal experience with AD and socio-demographic characteristics with emotional reactions to persons with AD. A total of 170 Israeli Arab adults, aged 40-85, were interviewed in person by the vignette methodology for their emotional reactions to people with AD, an issue not yet studied in this population. Their perceptions of dangerousness of persons with AD to themselves and to others, familiarity with the disease and perceptions of personal threat were also probed. Four dimensions of emotional reactions to a person with AD emerged: anxiety, aggressiveness, prosocial reactions and rejection. More positive than negative emotions were expressed. Levels of education and religiosity, and perceived dangerousness of the person with AD to others, were the main factors associated with emotional reactions. Culturally based educational programs should be developed to increase knowledge and reduce negative reactions, thus increasing the wellbeing of AD sufferers and their families.


Amylin, Aβ42, and Amyloid in Varicella Zoster Virus Vasculopathy Cerebrospinal Fluid and Infected Vascular Cells.
Varicella zoster virus (VZV) vasculopathy is characterized by persistent arterial inflammation leading to stroke. Studies show that VZV induces amyloid formation that may aggravate vasculitis. Thus, we determined if VZV central nervous system infection produces amyloid. Aβ peptides, amylin, and amyloid were measured in cerebrospinal fluid (CSF) from 16 VZV vasculopathy subjects and 36 stroke controls. To determine if infection induced amyloid deposition, mock- and VZV-infected quiescent primary human perineurial cells (qHPNCs), present in vasculature, were analyzed for intracellular amyloidogenic transcripts/proteins and amyloid. Supernatants were assayed for amyloidogenic peptides and ability to induce amyloid formation. To determine amylin's function during infection, amylin was knocked down with small interfering RNA and viral complementary DNA (cDNA) was quantitated. Compared to controls, VZV vasculopathy CSF had increased amyloid that positively correlated with amylin and anti-VZV antibody levels; Aβ40 was reduced and Aβ42 unchanged. Intracellular amylin, Aβ42, and amyloid were seen only in VZV-infected qHPNCs. VZV-infected supernatant formed amyloid fibrils following addition of amyloidogenic peptides. Amylin knockdown decreased viral cDNA. VZV infection increased levels of amyloidogenic peptides and amyloid in CSF and qHPNCs, indicating that VZV-induced amyloid deposition may contribute to persistent arterial inflammation in VZV vasculopathy. In addition, we identified a novel proviral function of amylin.


Managing the tension: From innovation to application in health professions education.
Innovations in education are essential for solving problems and introducing new ways of thinking. However, implementation of these innovations must take several factors into consideration, including the context, the environment, the stakeholders, the technology needed, the cost, the pace of implementation, appropriateness, and available resources. When these factors are not balanced and considered, tensions arise. This paper describes tensions in five major educational domains, namely curriculum, instruction, assessment, accreditation/regulation, and faculty development. For each of these domains, the typical problems are described, and solutions are presented to manage the tension.


A reproducible genetic transformation system for cultivated Phaseolus acutifolius (tepary bean) and its use to assess the role of arcelins in resistance to the Mexican bean weevil.
A reproducible Agrobacterium tumefaciens-mediated genetic transformation method that delivers fertile and morphologically normal transgenic plants was developed for cultivated tepary bean (Phaseolus acutifolius L. Gray). Factors contributing to higher transformation efficiencies include (1) a low initial concentration of bacteria coupled with a longer cocultivation period with callus, (2) an initial selection of callus on a medium containing low levels of the selectable agent, (3) omission of the selectable agent from the medium during callus differentiation to shoots and (4) the efficient conversion of transgenic shoots into fertile plants. All plants regenerated with this procedure (T0) were stably transformed, and the introduced foreign genes were inherited in a Mendelian fashion in most of the 33 independent transformants. Integration, stable transmission and high expression levels of the transgenes were observed in the T1 and/or T3 progenies of the transgenic lines. The binary transformation vectors contained the beta-glucuronidase reporter gene, the neomycin phosphotransferase II selectable marker gene and either an arcelin 1 or an arcelin 5 gene. Arcelins are seed proteins that are very abundant in some wild P. vulgaris L. genotypes showing resistance to the storage insect Zabrotes subfasciatus (Boheman) (Coleoptera, Bruchidae). Transgenic beans from two different cultivated P. acutifolius genotypes with high arcelin levels were infested with Z. subfasciatus, but they were only marginally less susceptible to infestation than the non-transgenic P. acutifolius. Hence, the arcelin genes tested here are not major determinants of resistance against Z. subfasciatus.


Impacts of cost functions on inverse lithography patterning.
For advanced CMOS processes, inverse lithography promises better patterning fidelity than conventional mask correction techniques due to a more complete exploration of the solution space. However, the success of inverse lithography relies highly on customized cost functions whose design and know-how have rarely been discussed. In this paper, we investigate the impacts of various objective functions and their superposition for inverse lithography patterning using a generic gradient descent approach. We investigate the most commonly used objective functions, which are the resist and aerial images, and also present a derivation for the aerial image contrast. We then discuss the resulting pattern fidelity and final mask characteristics for simple layouts with a single isolated contact and two nested contacts. We show that a cost function composed of a dominant resist-image component and a minor aerial-image or image-contrast component can achieve a good mask correction and contour targets when using inverse lithography patterning.


Mechanical and in vitro biological properties of uniform and graded Cobalt-chrome lattice structures in orthopedic implants.
Human bones are biological examples of functionally graded lattice capable to withstand large in vivo loading and allowing optimal stress distribution. Disruption of bone integrity may require biocompatible implants capable to restore the original bone structure and properties. This study aimed at comparing mechanical properties and biological behavior in vitro of uniform (POR-FIX) and graded (POR-VAR) Cobalt-chrome alloy lattice structures manufactured via Selective Laser Melting. In compression, the POR-VAR equivalent maximum stress was about 2.5 times lower than that of the POR-FIX. According to the DIC analysis, the graded lattice structures showed a stratified deformation associated to unit cells variation. At each timepoint, osteoblast cells were observed to colonize the surface and the first layer of both scaffolds. Cell activity was always significantly higher in the POR-VAR (p < 0.0005). In terms of gene expression, the OPG/RANKL ratio increased significantly over time (p < 0.0005) whereas IL1β and COX2 significantly decreased (7 day vs 1 day; p < 0.0005) in both scaffolds. Both uniform- and graded-porosity scaffolds provided a suitable environment for osteoblasts colonization and proliferation, but graded structures seem to represent a better solution to improve stress distribution between implant and bone of orthopedic implants.


Barriers to Providing VTE Chemoprophylaxis to Hospitalized Patients: A Nursing-Focused Qualitative Evaluation.
Venous thromboembolism (VTE) is a serious medical condition that results in preventable morbidity and mortality. The objective of this study was to identify nursing-related barriers to administration of VTE chemoprophylaxis to hospitalized patients. This was a qualitative study including nurses from five inpatient units at one hospital. Observations were conducted on five units to gain insight into the process for administering chemoprophylaxis. Focus group interviews were conducted with nurses and were audio-recorded, transcribed verbatim, and analyzed using the Theoretical Domains Framework to identify barriers to providing VTE chemoprophylaxis. We conducted 14 focus group interviews with nurses from five inpatient units to assess nurses' perceptions of barriers to administration of VTE chemoprophylaxis. The barriers identified included nurses' misconceptions that ambulating patients did not require chemoprophylaxis, nurses' uncertainty when counseling patients on the importance of chemoprophylaxis, and a lack of comparative data for nurses regarding their specific refusal rates. Multiple factors act as barriers to patients receiving VTE chemoprophylaxis. These barriers are often modifiable targets for quality improvement. There is a need to focus on behavior changes that will remove or minimize barriers and equip nurses to ensure administration of VTE chemoprophylaxis by engaging patients in their care.


Enabling Sensitive Phenotypic Profiling of Cancer-Derived Small Extracellular Vesicles Using Surface-Enhanced Raman Spectroscopy Nanotags.
Circulating cancer-derived small extracellular vesicles (EVs) are nanoscale membranous vesicles shed from cancer cells that are released into surrounding body fluids. Small EVs contain biomolecules associated with cancer such as DNA and proteins for cell-to-cell communication. Therefore, small EVs have been regarded as important cancer biomarkers for liquid biopsy-based cancer diagnosis and drug treatment monitoring. However, because of the high heterogeneity and low level of small EVs in body fluids, there is a high demand for sensitive detection and characterization of such vesicles at a molecular level. In this study, we have developed a sensitive and effective approach to simultaneously profile multiple protein biomarkers expressed on cancer-derived small EVs using surface-enhanced Raman spectroscopy (SERS) nanotags in a single test, without complex isolation steps. Rapid and multiplexed phenotypic profiling of small EVs is achieved by mixing specific detection antibody-coated SERS nanotags, filtered conditioned EV-suspended medium (conditioned EVs), and capture antibody (CD63)-conjugated magnetic beads to form a sandwich immunoassay. As a proof-of-concept demonstration, we applied this approach to characterize pancreatic cancer-derived EVs by simultaneously detecting three specific EV surface receptors including Glypican-1, epithelial cell adhesion molecules (EpCAMs), and CD44 variant isoform 6 (CD44V6). The sensitivity of this method was measured down to 2.3 × 106 particles/mL, which is more sensitive and shows higher multiplexing capability than most other reported EV profiling techniques, such as western blot, enzyme-linked immunosorbent assay, and flow cytometry. Furthermore, phenotypic profiling of small EVs from colorectal cancer and bladder cancer cell lines (SW480 and C3) was conducted and compared to those derived from pancreatic cancer (Panc-1), highlighting the significant difference in EV phenotypes for various cancer cell types suspended in both phosphate-buffered saline and plasma. Thus, we believe that this technology enables a comprehensive evaluation of small secreted EV heterogeneity with high sensitivity, offering strong potential for accurate noninvasive cancer diagnosis and monitoring of drug treatment. In addition, this assay provides point-of-care use because of the easy sample preparation and portable nature of the Raman spectrometer.


Bystander suppression of allergic airway inflammation by lung resident memory CD8+ T cells.
CD8+ memory T cells have recently been recognized as playing a key role in natural immunity against unrelated viral infections, a phenomenon referred to as "heterologous antiviral immunity." We now provide data that the cellular immunological interactions that underlie such heterologous immunity can play an equally important role in regulating T helper 2 immune responses and protecting mucosal surfaces from allergen-induced inflammation. Our data show that CD8+ T cells, either retained in the lung after infection with influenza virus, or adoptively transferred via the intranasal route can suppress allergic airway inflammation. The suppression is mediated by IFN-gamma, which acts to reduce the activation level, T helper 2 cytokine production, airways hyperresponsiveness, and migration of allergen-specific CD4+ T cells into the lung, whereas the systemic and draining lymph node responses remain unchanged. Of note, adoptive transfer of previously activated transgenic CD8+ T cells conferred protection against allergic airway inflammation, even in the absence of specific-antigen. Airway resident CD8+ T cells produced IFN-gamma when directly exposed to conditioned media from activated dendritic cells or the proinflammatory cytokines IL-12 and IL-18. Taken together these data indicate that effector/memory CD8+ T cells present in the airways produce IFN-gamma after inflammatory stimuli, independent of specific-antigen, and as a consequence play a key role in modifying the degree and frequency of allergic responses in the lung.


Real-time monitoring of hydrogen peroxide consumption in an oxidation reaction in molecular solvent and ionic liquids by a hydrogen peroxide electrochemical sensor.
An efficient electrochemical protocol to monitor hydrogen peroxide consumption during metal-catalyzed oxidation by using screen-printed electrodes modified with Prussian blue is presented. In particular, cyclooctene oxidation to cyclooctene oxide, catalyzed by a vanadium(V)-salophen complex (H(2)salophen=N,N'-o-phenylenebis(salicylideneimine)), in molecular and ionic media was tested. Initially, a protocol for batch analysis was developed for a monophasic system in acetonitrile, and subsequently, an in situ protocol was developed for a biphasic system of 1-butyl-3-methylimidazolium hexafluorophosphate/phosphate buffer. Calibration curves were performed in amperometric mode by applying -50 mV versus an Ag pseudo-reference. The calibration curve of hydrogen peroxide showed a linear correlation from 1 × 10(-6) up to 5 × 10(-3) mol L(-1) with satisfactory inter- and intra-electrode reproducibility (relative standard deviation (RSD) values of 5 and 13%, respectively, for the monophasic system and 11 and 13%, respectively, for the biphasic system). Kinetic studies to investigate the oxidation reaction for both the mono- and biphasic systems have been carried out in amperometric mode as well. Firstly, the decomposition of hydrogen peroxide was examined, which showed that, in 1-butyl-3-methylimidazolium hexafluorophosphate(,) it completely decomposed in 300 min, whereas in acetonitrile, in the same time frame, 20% of the initial amount was still active. In the presence of 1% of the catalyst the decomposition rate increased in both solvents. Finally, the complete oxidation of cyclooctene was followed and the effective conversion was determined. The developed protocols showed high reproducibility, with the advantage that the environmentally friendly biphasic system could also be recycled. The good analytical performance obtained, coupled with a short analysis time, the possibility of in-line automation and the use of ionic liquids instead of molecular solvents, made this system a very attractive choice for monitoring oxidative reactions.


Immunocytochemical mapping of 1B236, a brain-specific neuronal polypeptide deduced from the sequence of a cloned mRNA.
The 318-amino acid, carboxy-terminal sequence of the putative brain-specific polypeptide 1B236 was deduced from the nucleotide sequence of its cloned brain-specific mRNA. Antisera raised against selected synthetic peptide fragments of this protein were used to map the cellular location of the presumptive gene product in the brains of normal or colchicine-pretreated adult rats. Antisera directed against any of three C-terminally located, but nonoverlapping, nonhomologous, synthetic peptide segments (P5, P6, or P7) produced virtually identical maps of intensely immunoreactive neuropil staining. The immunoreactivity was distributed heterogeneously and was most pronounced within olfactory, somatosensory, and limbic systems, and was more modest in certain motor and auditory structures. In colchicine-pretreated rats, large, multipolar perikarya were observed within the amygdala, caudate-putamen, cingulate, parietal, and piriform cortices, as well as in particular diencephalic and pontine nuclei. Smaller immunoreactive neurons with more limited dendritic extensions were observed in the olfactory bulb, the cerebellar cortex, and the dorsal horn and intermediolateral cell columns of the spinal cord. No immunoreactivity was observed in visceral structures innervated by the autonomic nervous system or in non-neural tissues. In addition to the virtually superimposable maps produced by antisera to all three synthetic fragments selected from the C-terminus of 1B236, some uniquely reactive sites were seen. Antisera to the most N-terminal of the three synthetic immunogens (P5) were reactive with neurons of the medial trapezoid nucleus and in nerve terminals surrounding the deep cerebellar nuclei. Antisera against the most C-terminal synthetic immunogen (P7) were reactive with neurons of the paraventricular and supraoptic hypothalamic nuclei. These data demonstrate that the 1B236 protein is located within selected neuronal elements within functionally related cellular circuits established more formally by other methods. Our data show that protein 1B236-immunoreactive cells share at least the expression of this protein and suggest that these cells may also be related epigenetically or evolutionarily. These data, together with other subcellular, ultrastructural, and electrophysiological properties of 1B236, suggest that this protein could be considered as a prohormone capable of yielding several final candidate transmitter products.


Simultaneity of foetal heart rate acceleration and foetal trunk movement determined by foetal magnetocardiogram actocardiography.
We describe a new method of actocardiography, foetal magnetocardiogram (fMCG) actocardiography, which is based on the high sensitivity of the fMCG to foetal trunk movements. We demonstrate the efficacy of the method by applying it to assess the simultaneity of onset of foetal heart rate (FHR) acceleration and foetal trunk movement. The analysis was restricted to events for which the onset of FHR accelerations and foetal movements could be determined accurately, i.e. when FHR was stable and near the average quiescent level just prior to the acceleration. We found that FHR accelerations coincided with or preceded foetal movements nearly all the time. This supports the hypothesis of coordinated control of FHR accelerations and foetal movements more strongly than prior studies, based on other techniques. We also found that beat-to-beat FHR variability often decreased at or near the start of FHR accelerations and that this occurrence was an accurate marker of foetal movement onset, even when foetal movement onset lagged FHR accelerations.


Can We Define Termination Of Resuscitation Criteria In Out-Of-Hospital Hanging?
Survival rate of cardiac arrest due to hanging (H-CA) victims is low. Hence, this leads to the question of the utility of resuscitation in these patients. The objective was to investigate whether there are predictive criteria for survival with a good neurological outcome or predictive criteria for non-survival or survival with a poor neurological outcome enabling us to define the termination of resuscitation rules in these patients. Between July 1, 2011 and January 1, 2016, we included 1,689 out-of-hospital cardiac arrests due to hanging. We compared the characteristics of survivors with a good neurological outcome at day 30 with the others. The study population was mainly composed of males with a median age of 48 [37-60]. The overall survival was 2.1%, among which 48.6% had a good neurological outcome. Survivors benefited more often from immediate basic life support than the rest of the subjects, which was corroborated by the shorter no-flow durations. We did not record any difference in terms of advanced cardiac life support initiation frequency and technique between survivors with a good neurological outcome and the rest. Nevertheless, ACLS duration was longer in survivors with a good neurological outcome than in others. Basic life support (BLS) was the decisive criterion for 15/17 survivors. However, a detailed analysis showed 2 survivors presenting no BLS before the arrival of mobile medical teams and non-shockable rhythms who survived at day 30 with a good neurological outcome. These results lead us to consider that mobile medical team intervention and ACLS attempt are not futile, and the benefit justifies the cost. Thus, we cannot define any rule for the termination of resuscitation.


Correlation Decay in Fermionic Lattice Systems with Power-Law Interactions at Nonzero Temperature.
We study correlations in fermionic lattice systems with long-range interactions in thermal equilibrium. We prove a bound on the correlation decay between anticommuting operators and generalize a long-range Lieb-Robinson-type bound. Our results show that in these systems of spatial dimension D with, not necessarily translation invariant, two-site interactions decaying algebraically with the distance with an exponent α≥2D, correlations between such operators decay at least algebraically to 0 with an exponent arbitrarily close to α at any nonzero temperature. Our bound is asymptotically tight, which we demonstrate by a high temperature expansion and by numerically analyzing density-density correlations in the one-dimensional quadratic (free, exactly solvable) Kitaev chain with long-range pairing.


Effects of partial and complete replacement of added phosphates with encapsulated phosphates on lipid oxidation inhibition in cooked ground meat during storage.
The objective of this research was to investigate the influence of various levels (0.0, 0.1, 0.2, 0.3, 0.4, 0.5%) of added encapsulated polyphosphates (sodium tripolyphosphate; sodium pyrophosphate) combined with unencapsulated polyphosphate to total 0.5% on the inhibition of lipid oxidation in cooked ground meat (beef, chicken) during refrigerated storage (0, 1, 7 d). The use of sodium tripolyphosphate (encapsulated sodium tripolyphosphate, unencapsulated sodium tripolyphosphate) led to lower cooking loss compared to sodium pyrophosphate in both meat species (p < 0.05). Increasing encapsulated sodium tripolyphosphate up to 0.3% decreased cooking loss in ground beef (p < 0.05). Added encapsulated polyphosphate at 0.5% had the same effect on pH as 0.5% unencapsulated polyphosphate in the cooked ground beef and chicken. A higher accumulation of orthophosphate was determined in the samples with sodium tripolyphosphate compared to those with sodium pyrophosphate (p < 0.05). Inclusion of a minimum of 0.1% encapsulated polyphosphate decreased thiobarbituric acid reactive substances and lipid hydroperoxides on 7 d. Increasing encapsulated sodium tripolyphosphate and encapsulated sodium pyrophosphate up to 0.2% in beef decreased thiobarbituric acid reactive substances at 7 d. Addition of 0.4% encapsulated sodium tripolyphosphate and 0.3% encapsulated sodium pyrophosphate in chicken prevented any increase in TBARS during storage. Incorporating encapsulated sodium pyrophosphate at 0.3% inhibited lipid hydroperoxide formation in beef and chicken. The meat industry could achieve enhanced lipid oxidation inhibition by replacing some of the unencapsulated polyphosphate with encapsulated polyphosphate in their product formulations.


Identification of HLA-A*0201/-A*2402-restricted CTL epitope-peptides derived from a novel cancer/testis antigen, MCAK, and induction of a specific antitumor immune response.
Cancer immunotherapy is a potential therapeutic strategy, in addition to surgical treatment, radiotherapy, and chemotherapy. Cancer-specific immunotherapy, such as the MAGE peptide vaccine, has been utilized clinically. How-ever, there are inherent limits to the effectiveness of vaccinotherapy using a single antigen because of the expression frequency of cancer-specific antigens on tumor cells. Thus, identification of a new cancer-specific antigen is needed. In this study, we examined the possibility of using cancer-specific immunotherapy based upon mitotic centromere-associated kinesin (MCAK) which was previously identified as a novel cancer/testis antigen. To evaluate the feasibility of developing cancer immunotherapy using MCAK peptides, we studied HLA-A*0201 and *2402 as targets for CTLs in the context of HLA class I molecules. By using a peptide with a sequence of AINPELLQL (amino acid positions 63-71 in MCAK, HLA-A*0201) and FFEIYNGKL (amino acid positions 401-409 in MCAK, HLA-A*2402), CTL responses could be induced from unseparated PBMCs by stimulation of freshly isolated, peptide-pulsed PBMCs as antigen-presenting cells (APCs) and also by using interleukin-7 and keyhole limpet hemocyanin in primary culture. The induced CTLs could lyse HLA-A-*0201/*2402 colon and gastric cancer cells expressing MCAK, as well as the peptide-pulsed target cells, in an HLA class l, and CD8 restricted manner. The identification of the MCAK/HLA-A*0201 and *2402 peptides suggests the possibility of designing peptide-based immunotherapeutic approaches that might prove effective in treating patients with MCAK-positive cancer.


Mouse CSB protein is important for gene expression in the presence of a single-strand break in the non-transcribed DNA strand.
CSB protein is required for strand-specific repair of bulky DNA lesions in transcribed genes and mediates transcription recovery after exposure to DNA-damaging agents. We enzymatically generated DNA single-strand breaks (SSBs) with 3'-OH and 5'-phosphate termini in defined positions of a plasmid-borne gene and measured their effect on transcription in cell lines with different statuses of the Csb gene. A single SSB in the transcribed region of the gene caused significant decrease of gene expression. In all tested cell lines of mouse and human origin, a SSB in the transcribed DNA strand was less harmful for gene expression than a SSB situated in the opposing DNA strand. CSB deficiency exhibited no effect on the expression of the nicked DNA in human fibroblasts immortalised by SV40 large T-antigen but caused a very strong decrease of gene expression in spontaneously transformed mouse embryonic fibroblasts (MEFs). Compared to the corresponding CSB-proficient MEFs, the effect was on average 6.7-fold stronger for a defined SSB located in the non-transcribed DNA strand, but only 2.4-fold for a SSB in the transcribed strand and 1.7-fold for a SSB located in the non-genic region. At the same time, CSB deficiency did not compromise the overall efficiency of repair of SSBs generated by treatment of the cells with hydrogen peroxide. The gene expression data thus indicate that CSB prevents irreversible transcription failures at the sites of DNA damage, acting preferentially at SSBs located in the non-transcribed DNA strand of the transcribed genes. We further conclude that SSBs in the non-transcribed DNA strand are commonly more harmful for transcription than those situated in the transcribed strand.


Serum osteocalcin level is associated with the mortality in Chinese patients with Fibrodysplasia ossificans progressiva aged ≤18 years at diagnosis.
Fibrodysplasia ossificans progressiva (FOP) is an ultra-rare genetic disorder characterized by extraskeletal heterotopic ossification. It is well recognized that FOP can lead to a devastating condition of disability. However, the mortality rate of FOP patients in China and risk factors for mortality are still largely unclear. We conducted a retrospective research on a cohort of 65 cases of FOP patients in China from 2008 to 2018. We reviewed medical records of these FOP patients to retrieve information such as date of birth/death, gender, clinical features, genotypes and biochemical parameters and analyze the correlation of these parameters with the mortality. 92.3% (60/65 cases) patients were classic FOP patients, 3.1% (2/65 cases) were FOP-plus and 4.6% (3/65 cases) were FOP variants. 9 cases of this cohort were dead during the ten-year period, and the overall mortality rate was 13.8%. c.617G > A mutation was confirmed in all non-survivors. In FOP patients≤18 years at diagnosis, non-survivors demonstrated significantly lower blood osteocalcin and alkaline phosphatase levels compared with survivors (P < 0.05), and spearman correlation and logistic regression analysis indicated that serum osteocalcin and alkaline phosphatase levels were negatively correlated with the mortality. Furthermore, the receiver-operating characteristic curve analysis showed serum osteocalcin had the largest area under the curve of 0.855 among four biochemical parameters, and serum osteocalcin < 65.9 ng/ml displayed a good capacity to discriminate the non-survivors from survivors in FOP patients aged 18 years and younger at diagnosis. Our findings showed that the mortality rate of FOP was 13.8% in China. Serum OC level was negatively correlated with the mortality in Chinese FOP patients ≤18 years at diagnosis.


Transport of cations and anions across forestomach epithelia: conclusions from in vitro studies.
Secretion of saliva as well as absorptive and secretory processes across forestomach epithelia ensures an optimal environment for microbial digestion in the forestomachs. Daily salivary secretion of sodium (Na+) exceeds the amount found in plasma by a factor of 2 to 3, while the secretion of bicarbonate (HCO3-) is 6 to 8 times higher than the amount of HCO3- in the total extracellular space. This implies a need for efficient absorptive mechanisms across forestomach epithelia to allow for an early recycling. While Na+ is absorbed from all forestomachs via Na+/H+ exchange and a non-selective cation channel that shows increased conductance at low concentrations of Mg2+, Ca2+ or H+ in the luminal microclima and at low intracellular Mg2+, HCO3- is secreted by the rumen for the buffering of ingesta but absorbed by the omasum to prevent liberation of CO2 in the abomasum. Fermentation provides short chain fatty acids and ammonia (NH3) that have to be absorbed both to meet nutrient requirements and maintain ruminal homeostasis of pH and osmolarity. The rumen is an important location for the absorption of essential minerals such as Mg2+ from the diet. Other ions can be absorbed, if delivered in sufficient amounts (Ca2+, Pi, K+, Cl- and NH4+). Although the presence of transport mechanisms for these electrolytes has been described earlier, our knowledge about their nature, regulation and crosstalk has increased greatly in the last years. New transport pathways have recently been added to our picture of epithelial transport across rumen and omasum, including an apical non-selective cation conductance, a basolateral anion conductance, an apical H+-ATPase, differently expressed anion exchangers and monocarboxylate transporters.


Renoprotective activity of telmisartan versus pioglitazone on ischemia/reperfusion induced renal damage in diabetic rats.
Diabetes mellitus (DM) causes organ dysfunction and increases the sensitivity of organs to damages.To test this hypothesis, we used renal ischemia/reperfusion (I/R) experiment to evaluate the renoprotective activity of telmisartan versus pioglitazone on I/R induced renal damage in diabetic rats. Renal I/R was performed in both normal and diabetic rats. The protocol comprised ischemia for 45 minutes followed by the reperfusion for 24 hours and a treatment period of two weeks before induction of ischemia. Renal I/R in both control and diabetic rats induced marked renal dysfunction associated with a significant increase in the arterial pressure, tumor necrosis factor alpha (TNF-alpha) levels, and the malondialdehyde formation (MDA). The activities of the anti-oxidant enzymes such as reduced glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) were found to be decreased significantly compared to control rats. Diabetic animals that underwent renal I/R exhibited a significant increase in all the studied parameters with a reduction in the anti-oxidant enzymes as compared to non-diabetic rats. Histo-pathological studies confirm these results. Treatment with pioglitazone or telmisartan demonstrated a significant improvement in the reperfusion-induced renal injury in comparison with diabetic I/R group, without difference between the two treated groups. Therefore, the treatment with pioglitazone or telmisartan have the same corrective effect. Type 2 diabetes had exaggerated renal I/R injury in STZ-NAD induced diabetes. Telmisartan treatment is equieffective as pioglitazone in attenuating acute I/R-induced renal injury in diabetic rats by a modification in the oxidative stress and the inflammation.


Parallelized evolutionary learning for detection of biclusters in gene expression data.
The analysis of gene expression data obtained from microarray experiments is important for discovering the biological process of genes. Biclustering algorithms have been proven to be able to group the genes with similar expression patterns under a number of experimental conditions. In this paper, we propose a new biclustering algorithm based on evolutionary learning. By converting the biclustering problem into a common clustering problem, the algorithm can be applied in a search space constructed by the conditions. To further reduce the size of the search space, we randomly separate the full conditions into a number of condition subsets (subspaces), each of which has a smaller number of conditions. The algorithm is applied to each subspace and is able to discover bicluster seeds within a limited computing time. Finally, an expanding and merging procedure is employed to combine the bicluster seeds into larger biclusters according to a homogeneity criterion. We test the performance of the proposed algorithm using synthetic and real microarray data sets. Compared with several previously developed biclustering algorithms, our algorithm demonstrates a significant improvement in discovering additive biclusters.


K33-Linked Polyubiquitination of Coronin 7 by Cul3-KLHL20 Ubiquitin E3 Ligase Regulates Protein Trafficking.
Ubiquitin chains are formed as structurally distinct polymers via different linkages, and several chain types including K33-linkage remain uncharacterized. Here, we describe a role for K33-polyubiquitination in protein trafficking. We show that the Cullin 3 (Cul3) substrate adaptor KLHL20 is localized to the trans-Golgi network (TGN) and is important for post-Golgi trafficking by promoting the biogenesis of TGN-derived transport carriers. The Cul3-KLHL20 ubiquitin E3 ligase catalyzes a nondegradable, K33-linked polyubiquitination on coronin 7 (Crn7), which facilitates Crn7 targeting to TGN through a ubiquitin-dependent interaction with Eps15. Blockage of K33-chain formation, Crn7 ubiquitination, or disruption of Crn7-Eps15 interaction impairs TGN-pool F-actin assembly, a process essential for generating transport carriers. Enforced targeting of Crn7 to TGN bypasses the requirement of K33-ubiquitination for TGN-pool F-actin assembly and post-Golgi trafficking. Our study reveals a role of KLHL20-mediated K33-ubiquitination of Crn7 in post-Golgi transport and identifies a cellular recognition mechanism for this ubiquitin chain type.


Case 320.
A 7-year-old boy with a history of pleuropulmonary blastoma after resection 6 years prior and germline DICER1 mutation was being monitored by physicians at a multidisciplinary genetic predisposition clinic. He demonstrated no evidence of recurrent pleuropulmonary blastoma, and his renal US, chest radiographic, and ocular screening examination results remained normal. Per age-directed screening guidelines, he underwent thyroid US (Figs 1-3). He had no signs or symptoms of hyper- or hypothyroidism. Physical examination was notable for the absence of thyromegaly or palpable nodule. US at 12-month follow-up showed no change in size or appearance of the left lobe (not shown). However, at this time, the Thyroid Imaging Reporting and Data System (TI-RADS) classification scheme was applied to the stable left lobe finding. The findings were discussed at a multidisciplinary thyroid nodule conference, and the decision was made to bring the patient back for a short-term follow-up for limited unenhanced MRI without sedation (Fig 4). A diagnosis was made based on the follow-up imaging findings.


Emergency intubation for respiratory failure in Guillain-Barré syndrome.
The consequences of emergency intubation in Guillain-Barré syndrome are not known. To review data from patients admitted to the intensive care unit with recent diagnosis of Guillain-Barré syndrome and intubation for respiratory failure. The database of patients with Guillain-Barré syndrome admitted to the intensive care units during the past 2 decades was reviewed. Emergency intubation was defined as need for ventilation in a patient with sudden dyspnea, cyanosis, or respiratory arrest. Outcome and pulmonary morbidity were assessed by comparing these patients with patients intubated electively. Six patients were intubated for acute respiratory distress and 1 patient for respiratory arrest. Thirty-six patients were intubated electively. Prolonged weaning was twice as common in the emergent group as in the elective group; a larger sample size might have demonstrated statistical significance. One patient with respiratory arrest developed marked anoxic encephalopathy; in all others, no differences were found in mortality, pulmonary morbidity, or duration of ventilatory assistance. None of the emergency intubations occurred in the last 15 years of the study. Emergency intubation in Guillain-Barré syndrome is uncommon but, when associated with respiratory arrest, can lead to anoxic encephalopathy. Duration of ventilator use and pulmonary morbidity are not increased in these patients.


The final word on nutritional screening and assessment in older persons.
To provide an updated perspective of how nutritional screening and assessment in older persons should be performed and reasonably implemented in the near future. Although nutritional screening and assessment should be fast and easy procedures, there is increasing evidence that more time should be dedicated to them. This is probably an answer to the claim to a medicine being more preventive than curative. Increasing interest is currently given to healthy aging and nutritional status is more likely to be addressed for its implications on functional status and disability. Important prognostic conditions, such as frailty, sarcopenia, and cachexia, which are closely linked to the nutritional domain, are at the top of the agenda. Therefore, body composition is a key issue and functional status is suggested as primary endpoint of nutrition trials. In this scenario, there is also a rationale for systematic assessment of inflammation, protein intake, and vitamin D status as potential contributing factors to reduced muscle mass and function. A 'second-generation' multidimensional nutritional screening and assessment including the evaluation of body composition, frailty, sarcopenia, and cachexia could be hypothesized. Nutritional assessment should be also completed by the systematic evaluation of inflammation, protein intake, and vitamin D status.


Evaluation of Quantum II microbiology system for identification of gram-negative bacteria of veterinary origin.
The ability of a rapid, semiautomated bacterial identification system, the Quantum II microbiology system (Abbott Laboratories, Irving, Tex.), to accurately identify gram-negative bacteria from veterinary sources was evaluated. A total of 378 isolates were tested, including 298 organisms in the family Enterobacteriaceae and strains representing Acinetobacter sp., Aeromonas sp., Flavobacterium sp., Pasteurella multocida, Plesiomonas sp., and Pseudomonas spp. Of these isolates, 333 (88.1%) were correctly identified, 20 (5.3%) were not identified, 10 (2.6%) were incorrectly identified at the genus level, and 15 (4.0%) were incorrectly identified at the species level. The Quantum II system correctly identified 268 (89.9%) of the isolates of Enterobacteriaceae and 65 (81.3%) of the nonenteric isolates. P. multocida was not identified correctly, and some nonenteric gram-negative bacteria of clinical significance in veterinary medicine are not included in the data base. The Quantum II system provided an accurate identification system for isolates of Enterobacteriaceae but had limited usefulness for the identification of other gram-negative bacteria of clinical significance in veterinary medicine.


Rift Valley fever virus detection in susceptible hosts with special emphasis in insects.
Rift Valley fever phlebovirus (RVFV, Phenuiviridae) is an emerging arbovirus that can cause potentially fatal disease in many host species including ruminants and humans. Thus, tools to detect this pathogen within tissue samples from routine diagnostic investigations or for research purposes are of major interest. This study compares the immunohistological usefulness of several mono- and polyclonal antibodies against RVFV epitopes in tissue samples derived from natural hosts of epidemiologic importance (sheep), potentially virus transmitting insect species (Culex quinquefasciatus, Aedes aegypti) as well as scientific infection models (mouse, Drosophila melanogaster, C6/36 cell pellet). While the nucleoprotein was the epitope most prominently detected in mammal and mosquito tissue samples, fruit fly tissues showed expression of glycoproteins only. Antibodies against non-structural proteins exhibited single cell reactions in salivary glands of mosquitoes and the C6/36 cell pellet. However, as single antibodies exhibited a cross reactivity of varying degree in non-infected specimens, a careful interpretation of positive reactions and consideration of adequate controls remains of critical importance. The results suggest that primary antibodies directed against viral nucleoproteins and glycoproteins can facilitate RVFV detection in mammals and insects, respectively, and therefore will allow RVFV detection for diagnostic and research purposes.


Inflammatory Bowel Disease and Risk of Adverse Pregnancy Outcomes.
Existing data on pregnancy complications in inflammatory bowel disease (IBD) are inconsistent. To address these inconsistencies, we investigated potential associations between IBD, IBD-related medication use during pregnancy, and pregnancy loss, pre-eclampsia, preterm delivery, Apgar score, and congenital abnormalities. We conducted a cohort study in >85,000 Danish National Birth Cohort women who were pregnant in the period 1996-2002 and had information on IBD, IBD-related medication use (systemic or local corticosteroids, 5-aminosalicylates), pregnancy outcomes and potential confounders. We evaluated associations between IBD and adverse pregnancy/birth outcomes using Cox regression and log-linear binomial regression. IBD was strongly and significantly associated with severe pre-eclampsia, preterm premature rupture of membranes and medically indicated preterm delivery in women using systemic corticosteroids during pregnancy (hazard ratios [HRs] >7). IBD was also associated with premature preterm rupture of membranes in women using local corticosteroid medications (HR 3.30, 95% confidence interval [CI] 1.33-8.20) and with medically indicated preterm delivery (HR 1.91, 95% CI 0.99-3.68) in non-medicated women. Furthermore, IBD was associated with low 5-minute Apgar score in term infants (risk ratio [RR] 2.19, 95% CI 1.03-4.66). Finally, Crohn's disease (but not ulcerative colitis) was associated with major congenital abnormalities in the offspring (RR 1.85, 95% CI 1.06-3.21). No child with a congenital abnormality born to a woman with IBD was exposed to systemic corticosteroids in utero. Women with IBD are at increased risk of severe pre-eclampsia, medically indicated preterm delivery, preterm premature rupture of membranes, and delivering infants with low Apgar score and major congenital malformations. These associations are only partly explained by severe disease as reflected by systemic corticosteroid use.


Thymoquinone Augments Cyclophosphamide-Mediated Inhibition of Cell Proliferation in Breast Cancer Cells
Objective: Cancer chemotherapy at the recommended doses is largely associated with toxicity, and also it is not effective enough to reduce the advancement of the disease at lower doses. Thymoquinone (TQ) is an active compound derived from black seeds (Nigella sativa) which exhibits anticancer activities. The aim of the present study was to investigate the synergistic effect of TQ alone and in combination with cyclophosphamide (cyclo), and to unravel the role of TQ in fatty acid synthase (FASN) mediated molecular signaling in Her2 + and Her2- breast cancer cell lines. Methods: The effect of TQ on the growth of Her2+ SKBR-3 and Her2- MDA-231 breast cancer lines were evaluated as percent cell viability by cytotoxicity-based MTT assay. The analysis of cell cycle arrest was done through flowcytometry followed by Western blot and RT-PCR to detect signaling events in the cells. Results: The data showed that TQ-cyclo (0.5mM-10μM) combination significantly inhibited the proliferation through the 5.49% and 57.72% accumulation of cells in sub-G1 and G1 respectively as 12% cells were shifted from G2/M phase in Her2+ breast cancer cells. Similarly, TQ-cyclo (0.5mM-20μM) combination exhibited that the 16.6% cells were arrested in Sub-G1 and only 3.54% cells were remained in G2/M phase as it was 22.89% in DMSO control in Her-2- breast cancers cells. Though TQ alone or in combination with cyclo alleviated the PI3K/Akt signaling by downregulating the phosphorylation of Akt and upregulating the PTEN, no changes was observed in FASN and Her-2 as well in both type of cells. The significant decreased expression of cyclin D1 was found in TQ-cyclo combinations. Conclusion: The current findings suggested that TQ can alter the cell cycle progression and induce cell death independent of FASN mediated signaling. In terms of clinical perspective, the present study clearly showed that TQ can broadly augment the effect of cyclo in breast cancer cases irrespective of Her-2+ or Her-.


Bagging improves reproducibility of functional parcellation of the human brain.
Increasing the reproducibility of neuroimaging measurement addresses a central impediment to the advancement of human neuroscience and its clinical applications. Recent efforts demonstrating variance in functional brain organization within and between individuals shows a need for improving reproducibility of functional parcellations without long scan times. We apply bootstrap aggregation, or bagging, to the problem of improving reproducibility in functional parcellation. We use two large datasets to demonstrate that compared to a standard clustering framework, bagging improves the reproducibility and test-retest reliability of both cortical and subcortical functional parcellations across a range of sites, scanners, samples, scan lengths, clustering algorithms, and clustering parameters (e.g., number of clusters, spatial constraints). With as little as 6 min of scan time, bagging creates more reproducible group and individual level parcellations than standard approaches with twice as much data. This suggests that regardless of the specific parcellation strategy employed, bagging may be a key method for improving functional parcellation and bringing functional neuroimaging-based measurement closer to clinical impact.


Postnatal dietary omega-3 fatty acid supplementation rescues glucocorticoid-programmed adiposity, hypertension, and hyperlipidemia in male rat offspring raised on a high-fat diet.
Fetal glucocorticoid excess programs several adverse outcomes in adult offspring, many of which can be prevented by postnatal, dietary omega-3 (n-3) fatty acids. Here we tested 2 separate hypotheses: 1) a postnatal high-fat diet exacerbates the glucocorticoid-programmed phenotype; and 2) postnatal, dietary n-3 fatty acids rescue programmed outcomes, even in the presence of a high-fat diet challenge. Pregnant Wistar rat dams were either untreated or administered dexamethasone acetate (Dex; 0.5 μg/mL drinking water) from day 13 of pregnancy. Offspring were cross-fostered to untreated mothers and males were weaned onto a standard (Std), high-fat, low n-3 (HF), or high-fat, high n-3 (HFHn-3) diet. Prenatal Dex reduced birth weight (26%) and delayed puberty onset by 1.2 days, irrespective of postnatal diet. Prenatal Dex programmed increased blood pressure in adult offspring, an effect worsened by the postnatal HF diet. Supplementation with high n-3 fatty acids, however, prevented both the Dex and HF-induced increases in blood pressure. Prenatal Dex also programmed increased adiposity, plasma cholesterol, and plasma triglyceride levels at 6 months of age, particularly in those offspring raised on the HF diet. But again, each of these adverse outcomes was rescued by supplementation of the HF diet with n-3 fatty acids. In conclusion, the capacity of n-3 fatty acids to overcome adverse programming outcomes remains evident, even in the presence of a HF diet challenge.


Oral Sucrose for Pain Relief During Clubfoot Casting: A Double-Blinded Randomized Controlled Trial.
Idiopathic clubfoot treatment is treated by manipulation and casting utilizing the Ponseti technique which can make the infant fussy and irritable. The goal of this study was to determine which intervention could decrease this pain response in infants undergoing Ponseti casting for idiopathic clubfeet. Our hypothesis was that the administration of oral sucrose solution or milk would be the most effective in accomplishing that goal. We conducted a double-blinded randomized controlled trial at a tertiary pediatric orthopaedic center on 33 children (average age=17.94 d; SD=20.51 d) undergoing clubfoot manipulation and casting and their guardians. Each cast was considered a new event and was randomized to an oral 20% sucrose solution (S), water (W), or milk (M) in a bottle (breast or nonbreast). We assessed the Neonatal Infant Pain Scale (NIPS), heart rate, and oxygen saturation before, during, and after the casting. A total of 131 casts were randomized and 118 analyzed (37 M, 42 S, 39 W). Each child underwent an average of 3.97 casts (SD=1.74). There were no significant differences seen between the groups before casting in their mean NIPS score (M=2.2; SD=2.38, S=1.84, SD=2.18, W=1.61, SD=2.12). However during casting, mean NIPS score for both milk, 0.91 (SD=1.26, P=0.0005) and sucrose, 0.64 (SD=1.27, P<0.0001) were significantly less than water, 2.27 (SD=2.03) but not different from each other (P=0.33). Postcasting, the sucrose NIPS score, 0.69 (SD=1.53) continued to be significantly less than milk, 2.11 (SD=2.37, P=0.0065. There was no correlation between heart rate or oxygen saturation and NIPS. Sucrose solution and milk during Ponseti casting and manipulation were effective in decreasing the pain response in children undergoing manipulation and casting for clubfeet. The sucrose solution administration continued the pain relief into the postcasting period. In addition to the benefits of improving the patient experience during casting, a less irritable child may result in better casting. Level 1 evidence.


Back pain: pathogenesis, diagnosis and management.
Back pain is a common problem that may have physical and psychosocial consequences for the patient if not managed effectively. Assessment should aim to identify any underlying pathology so that targeted treatment can be provided. The nurse has a central role in giving the patient information about managing pain and offering support and reassurance if pain persists.


Obesity and polycystic ovary syndrome.
The aetiology of Polycystic Ovary Syndrome (PCOS) is complex and multifactorial. There is much evidence, however, to suggest that adipose tissue plays an important role in the development and maintenance of PCOS pathology. There is a close correlation between adiposity and symptom severity in women with PCOS, and even modest reductions in weight generally translate into significant improvements in menstrual regularity, fertility and hyperandrogenic features. This review article considers the various mechanisms that might underlie this link between excess adiposity and PCOS - including the effects of differential insulin sensitivity, abnormal steroid hormone metabolism and adipocytokine secretion. Greater attention to the therapeutic options available to reduce the impact of excess adiposity on ovarian and metabolic function is essential to the management of PCOS.


Adverse drug reactions in hospitalized cardiac patients: characteristics and risk factors.
Adverse drug reactions (ADRs) appear more frequently than actually reported and registered. The main goal of our work was to analyze risk factors, incidence and characteristics of ADRs in hospitalized cardiac patients. This prospective study included 200 patients, hospitalized at Cardiology Center of the Clinical Centre of Montenegro. ADRs were collected using specially designed questionnaire, based on the list of symptoms and signs that could point out to potential ADRs. Data from medical charts of patients, lab tests and other available parameters were observed and combined with the data from questionnaire. Severity of ADRs were assessed as serious or nonserious according to the World Health Organization criteria. Causality was assessed using the Naranjo probability scale. A total of 34% of all the patients experienced at least one ADR. The most common ADRs occurred as nervous system disorders, less frequent were cardiovascular disorders, while the immune system disorders were the rarest. Sixteen percent of all ADRs were characterized as serious, most often caused by carvedilol and amiodarone. The majority of patients (97.3%) recovered without consequences. The multivariate analysis showed independent significant associations between ADRs and age, gender, comorbidities and polypragmasia. ADRs represent a significant issue in hospitalized cardiac patients population. The most significant predictors for ADRs in observed population were age, comorbidity, number of medications used during hospitalization and patients' gender. Preventive measures such as pharmacotherapy rationalization and continual education of health care professionals could reduce the frequency of ADRs appearance in patients with detected risk factors.


Types of articles in Advances in Clinical and Experimental Medicine in 2021 and 2022: Editors' perspective.
The present editorial summarizes the last 2 calendar years of Advances in Clinical and Experimental Medicine (ACEM) publication (2021 and 2022). The specific aims were: 1) To clarify the classification of papers published in ACEM; 2) To present motivations behind choosing this classification; 3) To show how this classification is reflected in citations. Six categories of papers published in ACEM are presented: editorials, meta-analyses, reviews (including systematic reviews), multicenter studies, research-in-progress studies, and research letters; lack of clear definitions for editorials, research letters and research-in-progress studies is discussed. Thematic fields covered by all categories in 2021 and 2022 are presented and differences in this regard between 2021 and 2022 are highlighted. Reasons for not publishing case reports (CRs) are discussed, with some of the debate on this issue in medical literature summarized. The article type classification used in ACEM in only one of many possible solutions and may be modified in the future - it should be both clear for the authors and allow for orientation in the journal's content. The motivation for choosing the employed categories stem both from their position on the accepted levels of evidence in evidence-based medicine (EBM) and their potential to be cited.


Cationic AIEgen micelle-improved chemiluminescent H2O2 assay by integrating reactant approach and CRET.
The enhancement of chemiluminescence (CL) intensity is significant in the development of chemiluminescent detection systems with improved sensitivity. In this study, a cationic surfactant with an intrinsic aggregation-induced emission emitter (AIEgen) has been applied to boost the CL signal of the horseradish peroxidase-luminol-H2O2 system. The formed cationic AIEgen micelles enhance the CL signal in two ways: the electrostatic attraction-mediated enrichment and approach of reactants and the high CRET efficiency between excited luminol radicals and AIEgen in the surfactant backbone. As a result, strong CL intensity is produced. Rapid and sensitive H2O2 detection is realized through the proposed cationic AIEgen micelle-containing chemiluminescent system with a limit of detection of 100 nM. The favourable selectivity over other possible interferents including metal ions and anions is due to the specific chemical reaction. Practical H2O2 analysis of thawing water samples with high accuracy using the proposed chemiluminescent platform is realized and is consistent with standard methods.


Urinary incontinence in men with chronic obstructive pulmonary disease.
This study investigated urinary incontinence in men with chronic obstructive pulmonary disease (COPD). A total of 244 community-dwelling men (mean age 66.5 years) diagnosed with COPD within the past 4 years were recruited from six hospital outpatient departments in central Japan. The prevalence of urinary incontinence was 10% according to the International Consultation on Incontinence criterion. Urine leakage among the 24 incontinent men was typically a small amount (75%) and occurred once a week or less often (58%). Fifteen (63%) of them reported urge incontinence while only two men experienced stress incontinence. On average they had urine leakage for 2.5 (SD 2.3) years and the majority (n = 19, 79%) developed the condition after diagnosis of COPD. The finding of higher prevalence of urge incontinence challenges the conventional view that COPD is associated with stress incontinence due to high pressure coughs.


Pre-Exposure Prophylaxis for HIV Prevention: Safety Concerns.
Available evidence supports the efficacy of pre-exposure prophylaxis (PrEP) in decreasing the incidence of human immunodeficiency virus (HIV) infection among high-risk individuals, especially when used in combination with other behavioural preventive methods. Safety concerns about PrEP present challenges in the implementation and use of PrEP. The aim of this review is to discuss safety concerns observed in completed clinical trials on the use of PrEP. We performed a literature search on PrEP in PubMed, global advocacy for HIV prevention (Aids Vaccine Advocacy Coalition) database, clinical trials registry " http://www.clinicaltrials.gov " and scholar.google, using combination search terms 'pre-exposure prophylaxis', 'safety concerns in the use of pre-exposure prophylaxis', 'truvada use as PrEP', 'guidelines for PrEP use', 'HIV pre-exposure prophylaxis' and 'tenofovir' to identify clinical trials and literature on PrEP. We present findings associated with safety issues on the use of PrEP based on a review of 11 clinical trials on PrEP with results on safety and efficacy as at April 2016. We also reviewed findings from routine real-life practice reports. The pharmacological intervention for PrEP was tenofovir disoproxil fumarate/emtricitabine in a combined form as Truvada® or tenofovir as a single entity. Both products are efficacious for PrEP and seem to have a good safety profile. Regular monitoring is recommended to prevent long-term toxic effects. The main adverse effects observed with PrEP are gastrointestinal related; basically mild to moderate nausea, vomiting and diarrhea. Other adverse drug effects worth monitoring are liver enzymes, renal function and bone mineral density. PrEP as an intervention to reduce HIV transmission appears to have a safe benefit-risk profile in clinical trials. It is recommended for widespread use but adherence monitoring and real-world safety surveillance are critical in the post-marketing phase to ensure that the benefits observed in clinical trials are maintained in real-world use.


Intravitreal versus sub-tenon posterior triamcinolone injection in bilateral diffuse diabetic macular edema.
To compare the efficacy of intravitreal and sub-Tenon (ST) posterior triamcinolone injection in the treatment of bilateral diabetic diffuse macular edema (DDME). 28 eyes of 14 diabetic patients with bilateral DDME were included in this prospective randomized study. One eye of each individual received a sub-Tenon (ST) posterior triamcinolone injection (ST group), whereas the fellow eye received an intravitreal triamcinolone injection (IV group). All eyes were treated with supplementary focal laser after the injections. Cases were evaluated 1, 2, 3 and 6 months after the initial treatment. Both groups had visual improvement and macular edema reduction following the injections. This temporary effect was prominent for 24 weeks in the IV group and for 8 weeks in the ST group. Edema resorption and visual improvement were more pronounced in the IV group, although this difference was not statistically significant. No major complications were observed except for moderate and reversible intraocular pressure elevation in some cases. Both intravitreal and ST injections of triamcinolone appear to yield a significant visual recovery and prompt resolution of DDME. The beneficial outcome appears to be more longstanding with intravitreal injection.


Incisal changes and orthodontic stability.
Lateral cephalograms and study casts of 55 patients were evaluated to determine if any relationships exist among incisal positions and angulations, changes in positions and angulations, and long-term occlusal stability. No significant relationships could be found between long-term changes occurring in a number of commonly used incisal measurements and end-of-treatment incisal positions, changes in incisal positions during treatment, or long-term changes in the facial axis angle, ANB angle, or weighted PAR score. Long-term incisal changes occurring in individual patients were not necessarily associated with negative occlusal changes. Since incisal positions usually change in the long-term, it is suggested that the use of published norms or recommended absolute goals for end-of-treatment incisal positions be used more as general functional and esthetic clinical guides, rather than as predictors of stability.


Closed pulmonary valvotomy in infants under 6 months of age: report of 14 consecutive cases without mortality.
Fourteen infants, all under 6 months of age, underwent surgery for the relief of severe valvar pulmonary stenosis. A modified Brock (transinfundibular valvotomy) procedure was performed in all cases. Dilatation of the pulmonary valve with a mosquito clamp and biliary dilator is the basis of the modified operation. All infants survived the operation. In two patients there is residual, significant pulmonary stenosis, and in two additional cases pulmonary regurgitation is present.


Repurposing of the Fasciolicide Triclabendazole to Treat Infections Caused by Staphylococcus spp. and Vancomycin-Resistant Enterococci.
One approach to combat the increasing incidence of multidrug-resistant (MDR) bacterial pathogens involves repurposing existing compounds with known safety and development pathways as new antibacterial classes with potentially novel mechanisms of action. Here, triclabendazole (TCBZ), a drug originally developed to treat Fasciola hepatica (liver fluke) in sheep and cattle, and later in humans, was evaluated as an antibacterial alone or in combination with sub-inhibitory concentrations of polymyxin B (PMB) against clinical isolates and reference strains of key Gram-positive and Gram-negative bacteria. We show for the first time that in vitro, TCBZ selectively kills methicillin-sensitive and methicillin-resistant Staphylococcus aureus and Staphylococcus pseudintermedius at a minimum inhibitory concentration (MIC) range of 2-4 µg/mL, and vancomycin-resistant enterococci at a MIC range of 4-8 µg/mL. TCBZ also inhibited key Gram-negative bacteria in the presence of sub-inhibitory concentrations of PMB, returning MIC90 values of 1 µg/mL for Escherichia coli, 8 µg/mL for Klebsiella pneumoniae, 2 µg/mL for Acinetobacter baumannii and 4 µg/mL for Pseudomonasaeruginosa. Interestingly, TCBZ was found to be bacteriostatic against intracellular S. aureus but bactericidal against intracellular S. pseudintermedius. Additionally, TCBZ's favourable pharmacokinetic (PK) and pharmacodynamic (PD) profile was further explored by in vivo safety and efficacy studies using a bioluminescent mouse model of S. aureus sepsis. We show that repeated four-hourly oral treatment of mice with 50 mg/kg TCBZ after systemic S. aureus challenge resulted in a significant reduction in S. aureus populations in the blood to 18 h post-infection (compared to untreated mice) but did not clear the bacterial infection from the bloodstream, consistent with in vivo bacteriostatic activity. These results indicate that additional pharmaceutical development of TCBZ may enhance its PK/PD, allowing it to be an appropriate candidate for the treatment of serious MDR bacterial pathogens.


Time-Resolved Selective Electrochemical Sensing of Carbon Particles.
This work demonstrated a new method for electrochemical detection of carbon black particles based on impact electrochemistry that was capable of selective detection of carbon black from the insulating oxide particles. We systematically studied the electrochemical collision events with carbon black particle suspension solution (pH 7.0 phosphate buffer) at varying carbon black concentrations using a convective condition and a gold microelectrode. We evaluated the effect of bias potential on the number and magnitude of collision spikes by changing the applied potential in chronoamperometry experiments. Our results showed that the biased potential of +0.4 V was the most suitable potential among the tested potential biases. Current blips were observed in the amperometric i-t response, and the spike numbers scaled linearly with the concentration of carbon black particles in the range of 2.5-20 μM (i.e., mass/volume concentration of 0.03 to 0.24 mg L-1) with the lowest detection limit of 0.396 μM (i.e., mass/volume concentration of 0.00475 mg L-1). The selective detection of carbon particles in the presence of representative poorly conductive oxide particles in our experimental conditions was achieved. The sensing mechanism of the sensitive and selective detection of carbon black particles is proposed. This work provides the basis for the development of powerful electroanalytical methods and technologies for the detection and classification of carbon particles in varying environmental conditions such as coalmines, engineered carbon particle factories, and coal power plants.


Levodopa pharmacokinetics and motor performance during activities of daily living in patients with Parkinson's disease on individual drug combinations.
Pharmacokinetics and pharmacodynamics of levodopa were evaluated at a high-resolution level in a heterogeneous group of 10 patients with idiopathic Parkinson's disease during their normal daily activity. A physician and a nurse spent 10 hours with each patient from the first morning dose of levodopa during daily activities at home and at work. Plasma samples were obtained every 20 minutes for analysis of levodopa and 3-O-methyldopa by high-performance liquid chromatography. To assess clinical response, mobility was rated on every test occasion by patients and by investigators. Food and fluid intake and physical activity were also monitored. There was a large intra- and interindividual variability in the pharmacokinetics of levodopa regardless of the different drug combinations used. Mean plasma levodopa concentration ranged between 0.45 to 7.07 microg/mL and peak concentrations between 0.95 to 13.75 microg/mL. In 44 of 58 dosing events, an oral dose of levodopa was related to a peak in plasma concentration. Assessment of the clinical effects was more sensitive when given by patients than when given by the investigators. The fluctuations of the levodopa concentration in plasma had a clear effect on the clinical parameters assessed, even during early disease stages. Variation in levodopa concentration is the determining factor for motor fluctuations also in patients on clinically optimized combinations with dopamine agonists and enzyme inhibitors.


[Early discharge possible in acute upper gastrointestinal bleeding; new study results require adaptation of Dutch guideline].
Acute upper gastrointestinal bleeding is still an important emergency situation with an incidence of 48-62 per 100,000 adults per year in the Netherlands. An international multicentre prospective study evaluated different risk scoring systems for patients presenting with upper gastrointestinal bleeding. The authors showed that patients with a Glasgow Blatchford score of 0 or 1 could be safely discharged without endoscopy. Thirty-day all-cause mortality in this group was 0.4%. In the current Dutch guideline, there is no clear advice on management of low-risk patients. The new guideline is expected in 2017 and should include this clear advice of discharging low-risk patients (Glasgow Blatchford score ≤ 1). It can therefore be concluded that with optimal selection about 20% of all patients presenting to the emergency room with acute upper gastrointestinal bleeding can be discharged the same day without intervention.


Regulation of PTEN degradation and NEDD4-1 E3 ligase activity by Numb.
The critical tumor suppressor PTEN is regulated by numerous post-translational modifications including phosphorylation, acetylation and ubiquitination. Ubiquitination of PTEN was reported to control both PTEN stability and nuclear localization. Notably, the HECT E3-ligase NEDD4-1 was identified as the ubiquitin ligase for PTEN, mediating its degradation and down-stream events. However, the mechanisms how NEDD4-1 is regulated by up-stream signaling pathways or interaction with other proteins in promoting PTEN degradation remain largely unclear. In the present study, we identified that the adaptor protein Numb, which is demonstrated to be a novel binding partner of NEDD4-1, plays important roles in controlling PTEN ubiquitination through regulating NEDD4-1 activity and the association between PTEN and NEDD4-1. Furthermore, we provided data to show that Numb regulates cell proliferation and glucose metabolism in a PTEN-dependent manner. Overall, our study revealed a novel regulation of the well-documented NEDD4-1/PTEN pathway and its oncogenic behavior.


Lactate is an antidepressant that mediates resilience to stress by modulating the hippocampal levels and activity of histone deacetylases.
Chronic stress promotes depression in some individuals, but has no effect in others. Susceptible individuals exhibit social avoidance and anxious behavior and ultimately develop depression, whereas resilient individuals live normally. Exercise counteracts the effects of stress. Our objective was to examine whether lactate, a metabolite produced during exercise and known to reproduce specific brain exercise-related changes, promotes resilience to stress and acts as an antidepressant. To determine whether lactate promotes resilience to stress, male C57BL/6 mice experienced daily defeat by a CD-1 aggressor, for 10 days. On the 11th day, mice were subjected to behavioral tests. Mice received lactate before each defeat session. When compared with control mice, mice exposed to stress displayed increased susceptibility, social avoidance and anxiety. Lactate promoted resilience to stress and rescued social avoidance and anxiety by restoring hippocampal class I histone deacetylase (HDAC) levels and activity, specifically HDAC2/3. To determine whether lactate is an antidepressant, mice only received lactate from days 12-25 and a second set of behavioral tests was conducted on day 26. In this paradigm, we examined whether lactate functions by regulating HDACs using co-treatment with CI-994, a brain-permeable class I HDAC inhibitor. When administered after the establishment of depression, lactate behaved as antidepressant. In this paradigm, lactate regulated HDAC5 and not HDAC2/3 levels. On the contrary, HDAC2/3 inhibition was antidepressant-like. This indicates that lactate mimics exercise's effects and rescues susceptibility to stress by modulating HDAC2/3 activity and suggests that HDAC2/3 play opposite roles before and after establishment of susceptibility to stress.


Cell death and oxidative stress in gliomas.
In gliomas, apoptosis and necrosis are determined by a number of promoting and inhibiting factors including oxidative cell stress mediated by nitric oxide synthases (NOS) and reduced by superoxide dismutases. Therefore, in 46 gliomas (including astrocytomas, oligodendrogliomas, oligo-astrocytomas, and glioblastomas), the relationship of apoptosis and necrosis and the expression of apoptosis-promoting (p53, bax, Fas, Fas-L) and inhibiting (bcl-2) factors as well as of different isoforms of NOS (NOSb, NOSe, NOSi) and manganese superoxide dismutase (MnSOD) were studied. Apoptosis was measured in situ by the TUNEL method while expression profiles of apoptosis-related and oxidative stress-associated factors were determined by immunohistochemistry. As a defining criterion, necrosis was restricted to glioblastomas while apoptosis increased with tumour malignancy (P=0.017) in all types of gliomas. Glial tumour cells displayed upregulation of bax, Fas, Fas-L, p53, and bcl-2 but with no significant correlation with malignancy. There was also a strong expression of NOS isoforms with upregulation of NOSe in all and of NOSb and NOSi in nearly 50% of the tumour specimens but only NOSb expression correlated significantly with tumour malignancy (P=0.004). Likewise, MnSOD was strongly expressed in all gliomas but was not correlated with tumour grade. There was a wide variability of expression in each tumour type without significant correlation between apoptosis and expression of apoptosis-associated or oxidative stress-related factors indicating that the network of regulating factors may be too complex for clear associations.


Ceramicines U-Z from Chisocheton ceramicus and structure-antimalarial activity relationship study.
Ceramicines are a series of limonoids which were isolated from the barks of Malaysian Chisocheton ceramicus (Meliaceae), and were known to show various biological activity. Six new limonoids, ceramicines U-Z (1-6), with a cyclopentanone[α]phenanthrene ring system with a β-furyl ring at C-17 were isolated from the barks of C. ceramicus. Their structures were determined on the basis of the 1D and 2D NMR analyses, and their absolute configurations were investigated by CD spectroscopy. Ceramicine W (3) exhibited potent antimalarial activity against Plasmodium falciparum 3D7 strain with IC50 value of 1.2 µM. In addition, the structure-antimalarial activity relationship (SAR) of the ceramicines was investigated to identify substituent patterns that may enhance activity. It appears that ring B and the functional groups in the vicinity of rings B and C are critical for the antimalarial activity of the ceramicines. In particular, bulky ester substituents with equatorial orientation at C-7 and C-12 greatly increase the antimalarial activity.


Modification of existing maximum residue levels in various crops and evaluation of confirmatory data following the Article 12 MRL review for trifloxystrobin.
In accordance with Article 6 of Regulation (EC) No 396/2005, the applicant Bayer CropScience submitted two applications to the competent national authorities appointed in the United Kingdom and Greece to modify the existing maximum residue levels (MRLs) for the active substance trifloxystrobin in various crops and in sweet peppers/bell peppers, respectively. Moreover, in the application submitted to the United Kingdom, the applicant also included a request to evaluate the confirmatory data identified in the framework of the MRL review under Article 12 of Regulation (EC) No 396/2005 as not available. In the framework of the assessment process, these applications were re-allocated to the Netherlands. The data submitted in support of the requests were found to be sufficient to derive MRL proposals for sweet peppers/bell peppers, kales, escaroles, herbs and edible flowers, beans with pods, oat and chicory roots. Since the data gaps identified in the MRL review were not addressed, risk managers might consider lowering the tentative MRLs in passion fruits/maracujas and leafy brassica other than kales to the limit of quantification (LOQ), and the implementation of the Codex MRL (CXL) in the EU legislation for cucumbers and gherkins. The new intended use on witloof/Belgian endives was not adequately supported by residue data and therefore an MRL proposal cannot be derived. Adequate analytical methods for enforcement are available to control the residues of trifloxystrobin on the commodities under consideration. Based on the results of the risk assessment performed, considering the exposure to residues of trifloxystrobin and its metabolites CGA 321113 (apart for passion fruits), EFSA concluded that the short-term and long-term intake of residues resulting from the uses of trifloxystrobin under consideration is unlikely to present a risk to consumer health, except for escaroles. For the intended indoor use on escaroles, a risk from short-term intake cannot be excluded, hence a modification of the existing EU MRL is not proposed for this commodity. The consumer risk assessment shall be regarded as indicative and affected by uncertainties.


Circulating Tumor-Macrophage Fusion Cells and Circulating Tumor Cells Complement Non-Small-Cell Lung Cancer Screening in Patients With Suspicious Lung-RADS 4 Nodules.
Low-dose computed tomography (LDCT) screening of high-risk patients decreases lung cancer-related mortality. However, high false-positive rates associated with LDCT result in unnecessary interventions. To distinguish non-small-cell lung cancer (NSCLC) from benign nodules, in the present study, we integrated cellular liquid biomarkers in patients with suspicious lung nodules (lung cancer screening reporting and data system [Lung-RADS] 4). Prospectively, 7.5 mL of blood was collected from 221 individuals (training set: 90 nonscreened NSCLC patients, 74 high-risk screening patients with no/benign nodules [Lung-RADS 1-3], and 20 never smokers; validation set: 37 patients with suspicious nodules [Lung-RADS 4]). Circulating tumor cells (CTCs), CTC clusters, and tumor-macrophage fusion (TMF) cells were identified by blinded analyses. Screening patients underwent a median of two LDCTs (range, 1-4) with a median surveillance time of 30 (range, 11-50) months. In the validation set of 37 Lung-RADS 4 patients, all circulating cellular biomarker counts (P < .005; Wilcoxon test) and positivity rates were significantly higher in 23 biopsy-proven NSCLC patients (CTCs: 23 of 23 [100%], CTC clusters: 6 of 23 [26.1%], and TMF cells: 15 of 23 [65.2%]) than in 14 patients with biopsy-proven benign nodules (6 of 14 [42.9%], 0 of 14 [0%], and 2 of 14 [14.3%]). On the basis of cutoff values from the training set, logistic regression with receiver operating characteristic and area under the curve analyses demonstrated that CTCs (sensitivity: 0.870, specificity: 1.0, and area under the curve: 0.989) and TMF cells (0.652; 0.880; 0.790) complement LDCT in diagnosing NSCLC in Lung-RADS 4 patients. Cellular liquid biomarkers have a potential to complement LDCT interpretation of suspicious Lung-RADS 4 nodules to distinguish NSCLC from benign lung nodules. A future prospective, large-scale, multicenter clinical trial should validate the role of cellular liquid biomarkers in improving diagnostic accuracy in high-risk patients with Lung-RADS 4 nodules.


Dose-Specific Effects of Di-Isononyl Phthalate on the Endocannabinoid System and on Liver of Female Zebrafish.
Phthalates, used as plasticizers, have become a ubiquitous contaminant and have been reported for their potential to induce toxicity in living organisms. Among them, di-isononyl phthalate (DiNP) has been recently used to replace di(2-ethylhexyl) phthalate (DEHP). Nowadays, there is evidence that DiNP is an endocrine-disrupting chemical; however, little is known about its effects on the endocannabinoid system (ECS) and lipid metabolism. Hence, the aim of our study was to investigate the effects of DiNP on the ECS in zebrafish liver and brain and on hepatic lipid storage. To do so, adult female zebrafish were exposed to three concentrations (0.42 µg/L, 4.2 µg/L, and 42 µg/L) of DiNP via water for 3 weeks. Afterwards, we investigated transcript levels for genes involved in the ECS of the brain and liver as well as liver histology and image analysis, Fourier-transform infrared spectroscopy imaging, and measurement of endocannabinoid levels. Our results demonstrate that DiNP upregulates orexigenic signals and causes hepatosteatosis together with deregulation of the peripheral ECS and lipid metabolism. A decrease in the levels of ECS components at the central level was observed after exposure to the highest DiNP concentration tested. These findings suggest that replacement of DEHP with DiNP should be considered with caution because of observed adverse DiNP effects on aquatic organisms.


Volunteers trained in palliative care at the hospital: an original and dynamic resource.
Volunteers trained in palliative care are increasingly present in acute care units in general hospitals. Nevertheless, there still are few available data on this topic, especially concerning the integration of volunteers outside the palliative structures. Our present study aimed to describe the experience of volunteers trained in palliative care in the context of a primary care hospital. In particular, the difficulties and the benefits of this specific position were evaluated according to volunteers' own perceptions and words. We employed a qualitative method. Various aspects of the volunteer's role were explored by means of semistructured questions, addressing their activity, their motivations, and their feelings. Participants were volunteers (n = 19) trained in palliative care and working at a university hospital. After giving written consent, they completed the semistructured questionnaire at home. Content analysis was used to identify the main categories of answers and the principal themes reported by the volunteers. The main difficulties were related to uncertainty of the context. As every situation is different, volunteers could not define their role once and for all. However, they derived great satisfaction from their activity. A supporting frame and a good balance between constraints and autonomy were facilitating factors. Besides, the complexity related to the context contributed to make the position valuable and challenging. Integrating a voluntary service in a primary care hospital is partly based on active participation of the volunteers in developing their position in a more adequate way. In return, this relative autonomy implies a rigorous and supportive attitude from the institution.


Benign tumors in myotonic dystrophy type I target disease-related cancer sites.
Recent evidence showed that myotonic dystrophy type I (DM1) patients are at increased risk of certain cancers, but the risk of benign tumors is unknown. We compared the risk of benign tumors in DM1 patients with matched DM1-free individuals and assessed the association between benign tumors and subsequent cancers. We identified 927 DM1 patients and 13,085 DM1-free individuals matched on gender, birth-year, clinic, and clinic-registration year from the UK Clinical Practice Research Datalink, a primary care records database. We used Cox regression models for statistical analyses. DM1 patients had elevated risks of thyroid nodules (Hazard Ratio [HR] = 10.4; 95% Confidence Interval [CI] = 3.91-27.52; P < 0.001), benign tumors of the brain or nervous system (HR = 8.4; 95% CI = 2.48-28.47; P < 0.001), colorectal polyps (HR = 4.3; 95% CI = 1.76-10.41; P = 0.001), and possibly uterine fibroids (HR = 2.7; 95% CI = 1.22-5.88; P = 0.01). Pilomatricomas and salivary gland adenomas occurred almost exclusively in DM1 patients (Fisher's exact P < 0.001). The HR for colorectal polyps was elevated in DM1 males but not in females (HR = 8.2 vs. 1.3, respectively; P-heterogeneity < 0.001), whereas endocrine and brain tumors occurred exclusively in females. The data suggested an association between benign tumors and subsequent cancer in classic DM1 patients (HR = 2.7; 95% CI = 0.93-7.59; P = 0.07). Our study showed a similar site-specific benign tumor profile to that previously reported for DM1-associated cancers. The possible association between benign tumors and subsequent cancer in classic DM1 patients warrants further investigation as it may guide identifying patients at elevated risk of cancer. Our findings underscore the importance of following population-based screening recommendations in DM1 patients, for example, for colorectal cancer.


Antibiotic binding of STY3178, a yfdX protein from Salmonella Typhi.
The yfdX family proteins are known for long time to occur in various virulent bacteria including their multidrug resistant (MDR) strains, without any direct assigned function for them. However, yfdX protein along with other proteins involved in acid tolerance response is reported to be up regulated by the multidrug response regulatory system in E. coli. Hence, molecular and functional characterization of this protein is important for understanding of key cellular processes in bacterial cells. Here we study STY3178, a yfdX protein from a MDR strain of typhoid fever causing Salmonella Typhi. Our experimental results indicate that STY3178 is a helical protein existing in a trimeric oligomerization state in solution. We also observe many small antibiotics, like ciprofloxacin, rifampin and ampicillin viably interact with this protein. The dissociation constants from the quenching of steady state fluorescence and isothermal titration calorimetry show that ciprofloxacin binding is stronger than rifampin followed by ampicillin.