On calculating kerma, collision kerma and radiative yields.
To study the relationships between dose (D), kerma (K), and collision kerma ( K col ) in photon beams and to investigate total radiative yields for electrons and positrons as a function of energy. To do this accurately required calculating collision kerma directly as a function of position in a phantom and making changes to the EGSnrc package (including DOSRZnrc and g applications). Changes were made to the EGSnrc system to allow the user to distinguish events according to their initiating process, most importantly relaxation particles initiated by electron impact ionization as opposed to initiated by photons, especially those events depositing energy below energy cutoffs after relaxation events. Appropriate changes were made to the applications DOSRZnrc and g and a new application, DOSRZnrcKcol, was written. The modified codes are much more robust against changes in simulation parameters such as ECUT and AE and whether or not electron impact ionization is included in the simulation. The radiative yields for electrons generally differ from values in ICRU Report 371 (1984) which only account for radiative losses due to bremsstrahlung. The Monte Carlo calculated g(brems) values are generally greater than the ICRU 37 values due to energy-loss straggling. Plots of D, K and K col vs depth in megavoltage photon beams show that some "conventional wisdom" does not hold in general (e.g., D is not always greater than K past D max ) and in general at 10 cm depth it is found that D≈K and D > K col . The beam radius at which D / K col reaches its saturation value depends strongly on the threshold used to define reaching saturation and is generally greater than the radius for lateral charged particle equilibrium used in the TRS-483 Code of Practice for small beam dosimetry2 (Palmans et al, Med Phys. 2018;45:e1123-e1145). The changes to EGSnrc make kerma calculations more accurate but previous calculations with electron impact ionization turned off gave close to correct results. The application DOSRZnrcKcol makes calculating collision kerma more efficient and avoids various approximations used in the past although those approximations are shown to be justified. Including energy-loss straggling when calculating bremsstrahlung radiation yield increases the value. Fluorescence losses and annihilation in flight further increase the radiation yield of electrons and positrons. Results demonstrate the effects of EGSnrc using electron bremsstrahlung production cross sections for positrons and failing to model positron impact ionization.


Oligomeric forms of plant acetolactate synthase depend on flavin adenine dinucleotide.
Acetolactate synthase (ALS, EC 184.108.40.206) has been extracted and partially purified from etiolated barley shoots (Hordeum vulgare L.). Multiple forms of this enzyme were separated by gel filtration and/or anion-exchange chromatography using fast protein liquid chromatography. It could be demonstrated that these two species are in equilibrium, which strongly depends on the structural role of flavin adenine dinucleotide and pyruvate. With 50 micromolar of flavin adenine dinucleotide in the medium most of the ALS aggregates as a high molecular weight form (M(r) = 440,000), while 50 millimolar pyruvate facilitates dissociation into the smaller form (M(r) = 200,000). Data are presented to show that two enzymatically active forms are not isozymes but different oligomeric species or aggregates of the basic 58-kilodalton subunit of ALS. These different ALS species exhibit little difference in feedback inhibition by valine, leucine and isoleucine or in inhibition by the sulfonylurea herbicide chlorsulfuron. Both aggregation forms show a broad pH-optimum between 6.5 and 7. Furthermore, the affinity for pyruvate and the amount of directly-formed acetoin indicate similar properties of these separated ALS forms.


In Situ Production of Zoospores by Five Species of Phytophthora in Aqueous Environments for Use as Inocula.
The goal of this study was to develop a procedure that could be used to evaluate the potential susceptibility of aquatic plants used in constructed wetlands to species of Phytophthora commonly found in nurseries. V8 agar plugs from actively growing cultures of three or four isolates of Phytophthora cinnamomi, P. citrophthora, P. cryptogea, P. nicotianae, and P. palmivora were used to produce inocula. In a laboratory experiment, plugs were placed in plastic cups and covered with 1.5% nonsterile soil extract solution (SES) for 29 days, and zoospore presence and activity in the solution were monitored at 2- or 3-day intervals with a rhododendron leaf disk baiting bioassay. In a greenhouse experiment, plugs of each species of Phytophthora were placed in plastic pots and covered with either SES or Milli-Q water for 13 days during both summer and winter months, and zoospore presence in the solutions were monitored at 3-day intervals with the baiting bioassay and by filtration. Zoospores were present in solutions throughout the 29-day and 13-day experimental periods but consistency of zoospore release varied by species. In the laboratory experiment, colonization of leaf baits decreased over time for some species and often varied among isolates within a species. In the greenhouse experiment, bait colonization decreased over time in both summer and winter, varied among species of Phytophthora in the winter, and was better in Milli-Q water. Zoospore densities in solutions were greater in the summer than in the winter. Decreased zoospore activities for some species of Phytophthora were associated with prolonged temperatures below 13 or above 30°C in the greenhouse. Zoospores from plugs were released consistently in aqueous solutions for at least 13 days. This procedure can be used to provide in situ inocula for the five species of Phytophthora used in this study so that aquatic plant species can be evaluated for potential susceptibility.


The placenta as a barrier for toxic and essential elements in paired maternal and cord blood samples of South African delivering women.
Environmental toxicants such as metals may be detrimental to foetus and infant development and health because of their physiological immaturity, opportunistic and differential exposures, and a longer lifetime over which disease, initiated during pregnancy and in early life, can develop. The placental mechanisms responsible for regulation of absorption and excretion of elements during pregnancy are not fully understood. The aim of this paper is to assess the correlation for selected toxic and essential elements in paired whole blood samples of delivering women and cord blood, as well as to evaluate the placental permeability for selected elements. Regression analyses used to assess this correlation in 62-paired samples of maternal and cord whole blood of delivering women show that the concentrations of mercury, lead, cobalt, arsenic and selenium in maternal and cord blood differed statistically. Lead, cobalt, arsenic and selenium appear to pass the placental barrier by a diffusion mechanism. It was also found that the mercury levels in cord blood were almost double those of the mother, suggesting that the foetus may act as a filter for the maternal mercury levels during pregnancy. Transplacental transfer for arsenic and cobalt was 80% and 45%, respectively, suggesting that the placenta modulates the rate of transfer for these elements. Cadmium, manganese, copper and zinc levels did not show statistically significant correlations between two compartments (maternal versus cord whole blood). The study confirms that most of the toxic metals measured have an ability to cross the placental barrier.


Effect of seasonal variation on the composition and properties of raw milk destined for processing in the UK.
The composition and physical properties of raw milk from a commercial herd were studied over a 1 year period in order to understand how best to utilise milk for processing throughout the year. Protein and fat levels demonstrated seasonal trends, while minerals and many physical properties displayed considerable variations, which were apparently unrelated to season. However, rennet clotting time, ethanol stability and foaming ability were subject to seasonal variation. Many significant interrelationships in physico-chemical properties were found. It is clear that the milk supply may be more suited to the manufacture of different products at different times of the year or even on a day to day basis. Subsequent studies will report on variation in production and quality of products manufactured from the same milk samples described in the current study and will thus highlight potential advantages of seasonal processing of raw milk.


Meeting Patient and Professional Needs: Views of Stakeholders on a Training Initiative for DwSIs in Endodontics in London.
A pilot scheme was established across London to train NHS primary dental care practitioners to provide endodontic treatment of moderate difficulty. It was co-led by the former London Deanery (Health Education England: North West London) and local NHS commissioners. This research aimed to explore key stakeholders' perceptions about the purpose of the initiative, its advantages, disadvantages and future implications. Nineteen semi-structured interviews were conducted with stakeholders (commissioners and providers of the educational initiative; commissioners and providers of care, including trainees, principal dentists and specialists) involved in establishing, running and participating in the initiative and wider endodontic service provision in London. Interviews were based on a topic guide informed by the literature, and a workshop involving the London trainees. Interviews were recorded, transcribed and analysed using framework methodology. The project was perceived as supporting four key areas: addressing services, improving quality/outcomes, delivering education and enhancing professional status. There was evidence that dentists were harnessing health policy in facilitating 'reprofessionalisation' of dentistry with the creation of dentists with enhanced skills (DwSIs). Learning outcomes from the pilot were related to the accreditation of the participants, service tariffs, reimbursement for endodontic treatment on the NHS, and the need for continuity within and between services across the dental system. Uncertainty about funding and the changes within the NHS were among the concerns expressed regarding the future of the initiative. The findings of this research suggest that extending the skills of primary care practitioners may contribute to the reprofessionalisation of dentistry, which has much to contribute to patient care and the development of an integrated and accessible dental care system of quality, with improved outcomes for patients. The implications for health policy and further research are discussed.


"Reality" of near-death-experience memories: evidence from a psychodynamic and electrophysiological integrated study.
The nature of near-death-experiences (NDEs) is largely unknown but recent evidence suggests the intriguing possibility that NDEs may refer to actually "perceived," and stored, experiences (although not necessarily in relation to the external physical world). We adopted an integrated approach involving a hypnosis-based clinical protocol to improve recall and decrease memory inaccuracy together with electroencephalography (EEG) recording in order to investigate the characteristics of NDE memories and their neural markers compared to memories of both real and imagined events. We included 10 participants with NDEs, defined by the Greyson NDE scale, and 10 control subjects without NDE. Memories were assessed using the Memory Characteristics Questionnaire. Our hypnosis-based protocol increased the amount of details in the recall of all kind of memories considered (NDE, real, and imagined events). Findings showed that NDE memories were similar to real memories in terms of detail richness, self-referential, and emotional information. Moreover, NDE memories were significantly different from memories of imagined events. The pattern of EEG results indicated that real memory recall was positively associated with two memory-related frequency bands, i.e., high alpha and gamma. NDE memories were linked with theta band, a well-known marker of episodic memory. The recall of NDE memories was also related to delta band, which indexes processes such as the recollection of the past, as well as trance states, hallucinations, and other related portals to transpersonal experience. It is notable that the EEG pattern of correlations for NDE memory recall differed from the pattern for memories of imagined events. In conclusion, our findings suggest that, at a phenomenological level, NDE memories cannot be considered equivalent to imagined memories, and at a neural level, NDE memories are stored as episodic memories of events experienced in a peculiar state of consciousness.


Oxidative Chemical Stressors Alter the Physiological State of the Nasal Olfactory Mucosa of Atlantic Salmon.
The olfactory organs of fish have vital functions for chemosensory and defence. Though there have been some ground-breaking discoveries of their involvement in immunity against pathogens in recent years, little is known about how they respond to non-infectious agents, such as exogenous oxidants, which fish encounter regularly. To this end, we employed Atlantic salmon (Salmo salar) as a model to study the molecular responses at the nasal olfactory mucosa of a teleost fish when challenged with oxidants. Microarray analysis was employed to unravel the transcriptional changes at the nasal olfactory mucosa following two types of in vivo exposure to peracetic acid (PAA), a highly potent oxidative agent commonly used in aquaculture: Trial 1: periodic and low dose (1 ppm, every 3 days over 45 days) to simulate a routine disinfection; and Trial 2: less frequent and high dose (10 ppm for 30 min, every 15 days, 3 times) to mimic a bath treatment. Furthermore, leukocytes from the olfactory organ were isolated and exposed to PAA, as well as to hydrogen peroxide (H2O2) and acetic acid (AA)-the two other components of PAA trade products-to perform targeted cellular and molecular response profiling. In the first trial, microarrays identified 32 differentially expressed genes (DEG) after a 45-day oxidant exposure. Erythrocyte-specific genes were overly represented and substantially upregulated following exogenous oxidant exposure. In Trial 2, in which a higher dose was administered, 62 DEGs were identified, over 80% of which were significantly upregulated after exposure. Genes involved in immune response, redox balance and stress, maintenance of cellular integrity and extracellular matrix were markedly affected by the oxidant. All chemical stimuli (i.e., PAA, H2O2, AA) significantly affected the proliferation of nasal leukocytes, with indications of recovery observed in PAA- and H2O2-exposed cells. The migration of nasal leukocytes was promoted by H2O2, but not much by PAA and AA. The three chemical oxidative stressors triggered oxidative stress in nasal leukocytes as indicated by an increase in the intracellular reactive oxygen species level. This resulted in the mobilisation of antioxidant defences in the nasal leukocytes as shown by the upregulation of crucial genes for this response network. Though qPCR revealed changes in the expression of selected cytokines and heat shock protein genes following in vitro challenge, the responses were stochastic. The results from the study advance our understanding of the role that the nasal olfactory mucosa plays in host defence, particularly towards oxidative chemical stressors.


Chronic unpredictable stress influenced the behavioral but not the neurodegenerative impact of paraquat.
The impact of psychological stressors on the progression of motor and non-motor disturbances observed in Parkinson's disease (PD) has received little attention. Given that PD likely results from many different environmental "hits", we were interested in whether a chronic unpredictable stressor regimen would act additively or possibly even synergistically to augment the impact of the toxicant, paraquat, which has previously been linked to PD. Our findings support the contention that paraquat itself acted as a systemic stressor, with the pesticide increasing plasma corticosterone, as well as altering glucocorticoid receptor (GR) expression in the hippocampus. Furthermore, stressed mice that also received paraquat displayed synergistic motor coordination impairment on a rotarod test and augmented signs of anhedonia (sucrose preference test). The individual stressor and paraquat treatments also caused a range of non-motor (e.g. open field, Y and plus mazes) deficits, but there were no signs of an interaction (neither additive nor synergistic) between the insults. Similarly, paraquat caused the expected loss of substantia nigra dopamine neurons and microglial activation, but this effect was not further influenced by the chronic stressor. Taken together, these results indicate that paraquat has many effects comparable to that of a more traditional stressor and that at least some behavioral measures (i.e. sucrose preference and rotarod) are augmented by the combined pesticide and stress treatments. Thus, although psychological stressors might not necessarily increase the neurodegenerative effects of the toxicant exposure, they may promote co-morbid behaviors pathology.


Autoimmunity in uveitis.
Recent insights into the pathogenesis of immune-mediated diseases proposed a new classification, which includes autoimmune and auto-inflammatory diseases. The prevalence of specific autoimmune and auto-inflammatory diseases in uveitis and/or scleritis is not yet known. In this study, we examine the presence of systemic immune-mediated diseases in patients with uveitis and/or scleritis and put a special emphasis on autoimmune disorders by reporting on their clinical manifestations and visual prognosis. In this retrospective study, we reviewed data of 1327 patients presenting with uveitis and/or scleritis between January 2010 and July 2016 at the Erasmus Medical Center, Rotterdam, the Netherlands. All patients with noninfectious uveitis and/or scleritis were classified according to novel criteria for immune-mediated diseases. Various clinical data, including visual acuity (VA), of patients with uveitis of autoimmune origin were registered during 5-year follow-up. The origin of uveitis was in 5% (62/1327) autoimmune, in 15% (197/1327) auto-inflammatory and in 14% (180/1327) mixed autoimmune/auto-inflammatory. Patients with classical autoimmune connective tissue disease (N = 17) suffered mostly from rheumatoid arthritis and granulomatosis with polyangiitis and exhibited predominantly scleritis (53%). After 5 years of follow-up, none of the eyes of these patients developed legal blindness (VA of <0.1). The VA in patients with uveitis associated with autoimmune neuro-ophthalmological diseases (multiple sclerosis and neuromyelitis optica; N = 27) remained stable over time. Uveitis and scleritis of autoimmune origin were observed in 5% of the total series. The term autoimmune uveitis should not be used as a synonym for intraocular inflammation of noninfectious origin.


Research priorities in advanced heart failure: James Lind alliance priority setting partnership.
To determine research priorities in advanced heart failure (HF) for patients, carers and healthcare professionals. Priority setting partnership using the systematic James Lind Alliance method for ranking and setting research priorities. An initial open survey of patients, carers and healthcare professionals identified respondents' questions, which were categorised to produce a list of summary research questions; questions already answered in existing literature were removed. In a second survey of patients, carers and healthcare professionals, respondents ranked the summary research questions in order of priority. The top 25 unanswered research priorities were then considered at a face-to-face workshop using nominal group technique to agree on a 'top 10'. 192 respondents submitted 489 responses each containing one or more research uncertainty. Out-of-scope questions (35) were removed, and collating the responses produced 80 summary questions. Questions already answered in the literature (15) were removed. In the second survey, 65 questions were ranked by 128 respondents. The top 10 priorities were developed at a consensus meeting of stakeholders and included a focus on quality of life, psychological support, the impact on carers, role of the charity sector and managing prognostic uncertainty. Ranked priorities by physicians and patients were remarkably divergent. Engaging stakeholders in setting research priorities led to a novel set of research questions that might not have otherwise been considered. These priorities can be used by researchers and funders to direct future research towards the areas which matter most to people living with advanced HF.


Even mild hyperlactatemia is associated with increased mortality in critically ill patients.
The clinical significance of elevation of lactate levels within the reference range is not well studied. The objective of this study was to determine the best cutoff threshold for serum lactate within the reference range (0.01 to 2.00 mM) that best discriminated between survivors and nonsurvivors of critical illness and to examine the association between relative hyperlactatemia (lactate above the identified threshold) and mortality. This was a retrospective cohort study of adult patients admitted to the medical-surgical intensive care unit (ICU) of a tertiary care academic center. Youden index was calculated to identify the best lactate cutoff threshold that discriminated between survivors and nonsurvivors. Patients with lactate above the identified threshold were defined as having relative hyperlactatemia. Multivariate logistic regression, adjusting for baseline variables, was performed to determine the relationship between the above two ranges of lactate levels and mortality. In addition, a test of interaction was performed to assess the effect of selected subgroups on the association between relative hyperlactatemia and hospital mortality. During the study period, 2,157 patients were included in the study with mean lactate of 1.3 ± 0.4 mM, age of 55.1 ± 20.3 years, and acute physiology and chronic health evaluation (APACHE) II score of 22.1 ± 8.2. Vasopressors were required in 42.4%. Lactate of 1.35 mM was found to be the best cutoff threshold for the whole cohort. Relative hyperlactatemia was associated with increased hospital mortality (adjusted odds ratio (aOR), 1.60, 95% confidence interval (CI) 1.29 to 1.98), and ICU mortality (aOR, 1.66; 95% CI, 1.26 to 2.17) compared with a lactate level of 0.01 to 1.35 mM. This association was consistent among all examined subgroups. Relative hyperlactatemia (lactate of 1.36 to 2.00 mM) within the first 24 hours of ICU admission is an independent predictor of hospital and ICU mortality in critically ill patients.


Revisiting telegony: offspring inherit an acquired characteristic of their mother's previous mate.
Newly discovered non-genetic mechanisms break the link between genes and inheritance, thereby also raising the possibility that previous mating partners could influence traits in offspring sired by subsequent males that mate with the same female ('telegony'). In the fly Telostylinus angusticollis, males transmit their environmentally acquired condition via paternal effects on offspring body size. We manipulated male condition, and mated females to two males in high or low condition in a fully crossed design. Although the second male sired a large majority of offspring, offspring body size was influenced by the condition of the first male. This effect was not observed when females were exposed to the first male without mating, implicating semen-mediated effects rather than female differential allocation based on pre-mating assessment of male quality. Our results reveal a novel type of transgenerational effect with potential implications for the evolution of reproductive strategies.


The Beginnings of Metallurgy: A New Look: Arguments over diffusion and independent invention ignore the complex metallurgic crafts leading to iron.
In the 9 years since my first survey of early metallurgy appeared in this journal (65) metallurgy has become a major battleground between those who argue independent invention and those who argue diffusion in the evolution of urban civilization. In this new article I contend that: 1) The upland belt and debouching river valleys of southwestern Asia have a clear priority in the beginnings of copper metallurgy and extractive metallurgy generally, suggesting that the forces of urbanization contributing to the rise of metallurgy there were more massive, widespread, and better integrated than elsewhere. 2) The trend to polymetallism, against the background of pyrotechnology generally and the other important technologies of urbanism, established a necessary sequence to early metallurgy. This sequence was a prerequisite to the coming of the Iron Age, which was uniquely contained at first within the environment of Anatolia and the eastern Mediterranean and spread outward from there. 3) The course of metallurgy and possibly of the other urbanizing technologies can best be understood through a process of diffusion and multiple innovation interrelating metallurgical evolution over much of Eurasia, but with the area defined by the Black Sea, the Caspian Sea, the Red Sea, and the eastern Mediterranean as its center. Only in this fashion can we place metallurgy in its proper role as an important subsystem in the rise of civilization.


Fully automated digital holographic processing for monitoring the dynamics of a vesicle suspension under shear flow.
We investigate the dynamics of a vesicle suspension under shear flow between plates using DHM with a spatially reduced coherent source. Holograms are grabbed at a frequency of 24 frames/sec. The distribution of the vesicle suspension is obtained after numerical processing of the digital holograms sequence resulting in a 4D distribution. Obtaining this distribution is not straightforward and requires special processing to automate the analysis. We present an original method that fully automates the analysis and provides distributions that are further analyzed to extract physical properties of the fluid. Details of the numerical implementation, as well as sample experimental results are presented.


Immune responses of flounder Paralichthys olivaceus vaccinated by immersion of formalin-inactivated Edwardsiella tarda following hyperosmotic treatment.
The aim of the present study was to evaluate the effects of hyperosmotic immersion (HI) vaccination and determine the optimum hyperosmotic salinity for flounder Paralichthys olivaceus by investigating its immune responses following vaccination. Flounder were immersed in 1 of 3 hyperosmotic solutions at 50, 60 and 70‰ salinity, then transferred into 30‰ salinity normal seawater containing formalin-inactivated Edwardsiella tarda for vaccination (3 HI groups), or were immersed in normal seawater as direct immersion (DI group). The results showed that the percentages of surface membrane immunoglobulin-positive (sIg+) cells in peripheral blood leukocytes and spleen leukocytes induced by HI were significantly higher than that with DI (p < 0.05), and the 50‰ salinity group showed the strongest response among the HI groups, which reached peaks at Week 4. ELISA assay showed that the specific serum antibodies gradually increased after vaccination and reached peak at Day 32, and the fish treated with HI showed stronger antibody responses; among the HI groups, a significantly higher specific antibody level was detected in the 50‰ salinity group at Day 32 (p < 0.05). Similarly, the fish treated with HI showed higher specific mucosal antibody levels compared to the DI group, and the mucosal antibody showed a faster response, with peak time arriving 1 wk earlier than for the serum antibody. The relative percent survival (RPS) of flounder treated with HI at 50, 60 and 70‰ salinities were 79, 71 and 57% respectively, while this was 43% in the DI group. These results demonstrated that HI, especially the 50‰ salinity, could efficiently enhance the immune response of flounder and show higher RPS. This has significant value for immunological prevention of edwardsiellosis in flounder.


High-intensity red light suppresses melatonin.
Early studies on rodents indicated that the long-wavelength portion of the spectrum (orange- and red-appearing light) could influence circadian and neuroendocrine responses. Since then, both polychromatic and analytic action spectra in various rodent species have demonstrated that long-wavelength light is very weak, if not entirely inactive, for regulating neurobehavioral responses. Since testing of monochromatic light wavelengths above 600 nm is uncommon, many researchers have assumed that there is little to no effect of red light on the neuroendocrine or circadian systems. The aims of the following studies were to test the efficacy of monochromatic light above 600 nm for melatonin suppression in hamsters and humans. Results in hamsters show that 640 nm monochromatic light at 1.1 x 10(17) photons/cm2 can acutely suppress pineal melatonin levels. In normal healthy humans, equal photon density exposures of 1.9 x 10(18) photons/cm2 at 460, 630, and 700 nm monochromatic light elicited a significant melatonin suppression at 460 nm and small reductions of plasma melatonin levels at 630 and 700 nm. These findings are discussed relative to the possible roles of classical visual photoreceptors and the recently discovered intrinsically photosensitive retinal ganglion cells for circadian phototransduction. That physiology, and its potential for responding to red light, has implications for domestic applications involving animal care, the lighting of typical human environments, and advanced applications such as space exploration.


Arabidopsis ethylene signaling pathway.
In plants, ethylene gas functions as a potent endogenous growth regulator. In the model system Arabidopsis thaliana, the molecular mechanisms that underlie perception and transduction of the ethylene signal to the nucleus, where the transcription of hundreds of genes is altered, are being elucidated. In the current view, ethylene is sensed by a family of five receptors that show similarity to the bacterial two-component histidine kinases, and in plants function as negative regulators of the pathway. Binding of the ethylene gas turns off the receptors, resulting in the inactivation of another negative regulator of ethylene signaling, CTR1, a Raf-like protein kinase that directly interacts with the receptors. EIN2, a protein of unknown biochemical activity that functions as a positive regulator of the pathway, acts downstream of CTR. Derepression of EIN2 by ethylene upon disabling of the receptors and CTR1 leads to the activation of EIN3 and EIN3-like transcription factors. In the absence of ethylene, the levels of EIN3 protein are extremely low because of the function of two F-box-containing proteins, EBF1 and EBF2, that target EIN3 for proteosome-mediated degradation. In the presence of ethylene, the EIN3 protein accumulates in the nucleus and initiates a transcriptional cascade, resulting in the activation and repression of hundreds of genes. To date, the only empirically demonstrated direct target of EIN3 is the APETALA2 (AP2)-domain-containing transcription factor gene ERF1. The coregulation of ERF1 by another plant hormone, jasmonic acid, illustrates how a transcriptional cascade could be utilized in a combinatorial fashion to generate a large diversity of responses using a limited number of input signals. As new components and points of intersection with other pathways are identified, the Connections Map will be updated.


Expedited Investigation of Powder Caking Aided by Rapid 3D Prototyping of Testing Devices.
Powder caking can dramatically affect powder handling and downstream production processes. Understanding the key factors that contribute to bulk powder caking is crucial. This article introduces the Hirschberg caking device (HCD), which is a 3D-printed device allowing for parallel testing of powder caking in a cylindrical geometry. In the HCD setup, the powder sample is stored in controlled conditions in the sample holder. On removal of the sample holder, the caked powder will remain in the shape determined by the sample geometry while the remaining powder will fall down. Caking indices can be calculated based on image analysis and weight measurement. The results obtained for the caking of lactose monohydrate with the HCD were in good agreement with the results obtained by a ring shear tester. In addition, a strain tester was used to measure the strength of the formed cakes. Using this approach, critical storage conditions and the required concentration of a given anticaking agent (talc) for lactose monohydrate could be identified. This work demonstrates the potential of rapid prototyping in powder characterization by introducing a fast and affordable approach for exploring and trouble-shooting powder caking.


The effect of tea consumption on oxidative stress in smokers and nonsmokers.
While the anticarcinogenic effects of tea in animal models have been reported by several groups, human epidemiological studies examining tea consumption and cancer prevention have produced equivocal results. The beneficial properties of tea to human health may be related to the antioxidant properties of tea components. However, little evidence has been provided that tea consumption can either increase the antioxidant capacity or decrease oxidative stress in humans. In the present study, the effects of tea treatment (green tea) on biomarkers of oxidative stress were investigated in smokers and nonsmokers in two volunteer study groups (one in China and the other in United States). Green tea consumption in both study groups decreased oxidative DNA damage (8-OHdG in white blood cells and urine), lipid peroxidation (MDA in urine), and free radical generation (2, 3-DHBA in urine) in smokers. Nonsmokers (US study group) also exhibited a decrease in overall oxidative stress.


Molecular architecture of the PBP2-MreC core bacterial cell wall synthesis complex.
Bacterial cell wall biosynthesis is an essential process that requires the coordinated activity of peptidoglycan biosynthesis enzymes within multi-protein complexes involved in cell division (the "divisome") and lateral wall growth (the "elongasome"). MreC is a structural protein that serves as a platform during wall elongation, scaffolding other essential peptidoglycan biosynthesis macromolecules, such as penicillin-binding proteins. Despite the importance of these multi-partite complexes, details of their architecture have remained elusive due to the transitory nature of their interactions. Here, we present the crystal structures of the soluble PBP2:MreC core elongasome complex from Helicobacter pylori, and of uncomplexed PBP2. PBP2 recognizes the two-winged MreC molecule upon opening of its N-terminal region, revealing a hydrophobic zipper that serves as binding platform. The PBP2:MreC interface is essential both for protein recognition in vitro and maintenance of bacterial shape and growth. This work allows visualization as to how peptidoglycan machinery proteins are scaffolded, revealing interaction regions that could be targeted by tailored inhibitors.Bacterial wall biosynthesis is a complex process that requires the coordination of multiple enzymes. Here, the authors structurally characterize the PBP2:MreC complex involved in peptidoglycan elongation and cross-linking, and demonstrate that its disruption leads to loss of H. pylori shape and inability to sustain growth.


Hydrogeologic controls on episodic H2 release from precambrian fractured rocks--energy for deep subsurface life on earth and mars.
Dissolved H(2) concentrations up to the mM range and H(2) levels up to 9-58% by volume in the free gas phase are reported for groundwaters at sites in the Precambrian shields of Canada and Finland. Along with previously reported dissolved H(2) concentrations up to 7.4 mM for groundwaters from the Witwatersrand Basin, South Africa, these findings indicate that deep Precambrian Shield fracture waters contain some of the highest levels of dissolved H(2) ever reported and represent a potentially important energy-rich environment for subsurface microbial life. The delta (2)H isotope signatures of H(2) gas from Canada, Finland, and South Africa are consistent with a range of H(2)-producing water-rock reactions, depending on the geologic setting, which include both serpentinization and radiolysis. In Canada and Finland, several of the sites are in Archean greenstone belts characterized by ultramafic rocks that have under-gone serpentinization and may be ancient analogues for serpentinite-hosted gases recently reported at the Lost City Hydrothermal Field and other hydrothermal seafloor deposits. The hydrogeologically isolated nature of these fracture-controlled groundwater systems provides a mechanism whereby the products of water-rock interaction accumulate over geologic timescales, which produces correlations between high H(2) levels, abiogenic hydrocarbon signatures, and the high salinities and highly altered delta (18)O and delta (2)H values of these groundwaters. A conceptual model is presented that demonstrates how periodic opening of fractures and resultant mixing control the distribution and supply of H(2) and support a microbial community of H(2)-utilizing sulfate reducers and methanogens.


Pathophysiology and time course of silent myocardial ischaemia during mental stress: clinical, anatomical, and physiological correlates.
To define the prevalence and pathophysiology of myocardial ischaemia induced by mental stress in patients with coronary artery disease and exercise inducible ischaemia, and to determine the correlation between the severity of coronary artery disease and ischaemia induced by speech. Prospective cohort study. Tertiary care academic institution. PATIENTS AND PROTOCOL: 47 patients with coronary artery disease and 20 normal controls were studied using standardised exercise and mental stress. The ambulatory nuclear vest provided continuous measures of left ventricular ejection fraction and relative volume changes: an ischaemic response to mental stress was defined as a decrease in ejection fraction of > or = 5% for > or = 60 s. Severity of coronary artery disease was assessed by the extent of thallium reversibility on exercise testing and the severity of angiographic disease. 23 (49%) of 47 patients with coronary artery disease had an ischaemic response to mental stress which occurred early, was sustained throughout the task and associated with an increase in end systolic volume. In contrast, the pattern of left ventricular response in the remaining 24 patients (51%) resembled that in the normal controls. Patients with mental stress induced ischaemia tended to have greater severity of coronary disease (mean (SD) total number of diseased vessels 1.9 (0.8) v 1.4 (0.9), P = 0.07), more frequent exercise induced angina (17/23 v 7/24, P = 0.003) and lower increases in heart rate (36 (11) v 49 (23) beats per min, P = 0.023) and systolic blood pressure (32 (19) v 45 (18) mm Hg, P = 0.03) during exercise. Left ventricular responses to speech and exercise were compared in the 23 patients with mental stress induced ischaemia: mental stress was associated with a greater decrease in ejection fraction at comparable increases in rate pressure product (-6.5 (6.3)% v 4.7 (11.2)%, P = 0.0001). These findings suggest that mental stress induction of myocardial ischaemia is common in patients with stable coronary artery disease. Susceptible patients may have more functionally severe coronary disease. The time course, pattern, and haemodynamic features of mental stress induced ischaemia suggest a dynamic decrease in coronary supply.


The DNA-based human karyotype.
Image cytometry and computer analysis are used to determine the relative DNA content and the DNA-based centromeric index of the 24 chromosomes of the human karyotype. A two-step procedure is used. Chromosomes of cells in metaphase first are stained with quinacrine and identified visually by their fluorescent Q-band patterns. They then are stained for DNA using gallocyanin-chrome alum. The chromosome images are scanned and recorded as digital values of optical density by an CYDAC image cytometric microscope system, CYDAC. The digital images are processed by computer to measure for each chromosome the relative DNA stain contents of the whole chromosome and of the p and q arms and the DNA-based centromeric index. About ten cells are analyzed for each of the donors, who are phenotypically normal men and women. The chromosome measurements are pooled by chromosome type for each donor and are compared among donors. The means of the chromosome measurements give the DNA-based human karyotype. Analysis of the DNA-based data shows that some chromosomes or portions of chromosomes vary significantly among donors. These variants do not correlate with detectable morphologic polymorphisms, such as Q- or C-band variants; thus they represent new and otherwise undetectable chromosome polymorphisms whose genetic basis and clinical significance are yet to be determined.


Pharmacological interventions in the treatment of the acute effects of cannabis: a systematic review of literature.
Cannabis intoxication is related to a number of physical and mental health risks with ensuing social costs. However, little attention has been given to the investigation of possible pharmacological interactions in this condition. To review the available scientific literature concerning pharmacological interventions for the treatment of the acute effects of cannabis. A search was performed on the Pubmed, Lilacs, and Scielo online databases by combining the terms cannabis, intoxication, psychosis, anxiety, and treatment. The articles selected from this search had their reference lists checked for additional publications related to the topic of the review. The reviewed articles consisted of case reports and controlled clinical trials and are presented according to interventions targeting the physiological, psychiatric, and cognitive symptoms provoked by cannabis. The pharmacological interventions reported in these studies include: beta-blockers, antiarrhythmic agents, antagonists of CB-1 and GABA-benzodiazepine receptors, antipsychotics, and cannabidiol. Although scarce, the evidence on pharmacological interventions for the management of cannabis intoxication suggests that propanolol and rimonabant are the most effective compounds currently available to treat the physiological and subjective effects of the drug. Further studies are necessary to establish the real effectiveness of these two medications, as well as the effectiveness of other candidate compounds to counteract the effects of cannabis intoxication, such as cannabidiol and flumazenil.


Early development of therapeutic biologics--pharmacokinetics.
Modern biologics are biotechnology-derived pharmaceuticals. They are mostly used for diagnosis, prevention and treatment of serious and chronic diseases. Today, therapeutic biologics range from traditional biologics like blood and blood components, fractionated blood products, and antitoxins to modern biologics such as monoclonal antibodies, cytokines (e.g. interferon, interleukine), tissue growth factors, vaccines directed against non-infectious disease targets, and gene transfer products. Chemical as well as pre-clinical development are major challenges for biologics due to their different physicochemical properties (mostly protein structure) compared to small molecules. They demonstrate much more complex pharmacokinetic behaviour, which strongly influences their pre-clinical testing strategy. Biologics are often highly species-specific in action and immunogenic in test animal species and humans. Immunogenicity of therapeutic biologics may influence their pharmacokinetic behaviour as well as pharmacodynamics and toxicity. Biologics are frequently regulated by different procedures compared to small molecules. New guidances are evolving which reflect the rapid development of new technologies in this field. Bioanalytical method development and validation is a prerequisite not exclusively for pharmacokinetic studies but for the whole pre-clinical and clinical development. Due to their unique properties, different kinds of bioanalytical assays (mass assays, activity assays, immunogenicity assays) are necessary in early development of biologics.


Studying ZIF-8 SURMOF Thin Films with a Langatate Crystal Microbalance: Single-Component Gas Adsorption Isotherms Measured at Elevated Temperatures and Pressures.
Characterizing the adsorption behavior of thin films at technical operation ranges is highly relevant in order to obtain appropriate experimental parameters, as well as to evaluate their possible integration in multiple applications. Nonetheless, gathering such experimental data is not a trivial task. In the case of metal-organic framework thin films, a particular interesting class of highly porous coatings, determination of adsorption isotherms at elevated temperatures and pressures is not commonly reported. Using a custom-designed langatate crystal microbalance following the principle for piezoelectric microbalances, but under harsher conditions, allowed us to present adsorption equilibrium measurements on a zeolitic imidazolate framework-8 (ZIF-8) surface-mounted metal-organic framework (SURMOF) thin film at temperatures between 35 and 100 °C and at a pressure range up to 10 bars. The layer-by-layer liquid-phase epitaxy approach was used to deposit ZIF-8 SURMOF thin films on langatate crystals. X-ray diffraction, infrared reflection absorption spectroscopy, and scanning electron microscopy combined with energy dispersive X-ray mapping were used to gather deeper information on the ZIF-8 thin film growth. Single-component isotherms were collected for different gases of practical interest, that is, CO2, C2H6, C2H4, C3H8, and C3H6. The measurements were obtained at 35, 50, 75, and 100 °C at a gauge pressure range up to 10 bars. The collected data showed a clear preferential adsorption of ethane over ethylene, whereas only a slight difference was found between propane and propylene. Additionally, the experimental data allowed for the determination of adsorption equilibrium constants and saturation loadings of the ZIF-8 material as a thin SURMOF film, which is of great importance for its integration in multiple applications, such as sensor devices or in membrane-based separation.


Neurofeedback in healthy elderly human subjects with electroencephalographic risk for cognitive disorder.
In normal elderly subjects, the best electroencephalogram (EEG)-based predictor of cognitive impairment is theta EEG activity abnormally high for their age. The goal of this work was to explore the effectiveness of a neurofeedback (NFB) protocol in reducing theta EEG activity in normal elderly subjects who present abnormally high theta absolute power (AP). Fourteen subjects were randomly assigned to either the experimental group or the control group; the experimental group received a reward (tone of 1000 Hz) when the theta AP was reduced, and the control group received a placebo treatment, a random administration of the same tone. The results show that the experimental group exhibits greater improvement in EEG and behavioral measures. However, subjects of the control group also show improved EEG values and in memory, which may be attributed to a placebo effect. However, the effect of the NFB treatment was clear in the EG, although a placebo effect may also have been present.


Targeting the Neurokinin-1 Receptor Compromises Canonical Wnt Signaling in Hepatoblastoma.
The substance P (SP)/NK-1 receptor (NK1R) complex represents an intriguing anticancer target for a variety of tumors, including hepatoblastoma (HB). Therefore, NK1R antagonists, such as the clinical drug aprepitant, recently have been proposed as potent anticancer agents. However, very little is known regarding the molecular basis of NK1R inhibition in cancer. Using reverse phase protein array, Western blot, Super TOP/FOP, confocal microscopy, and sphere formation ability (SFA) assays, we identified the AKT and Wnt signaling pathways as the key targets of aprepitant in three human HB cell lines (HepT1, HepG2, and HuH6). Following NK1R blockage, we observed decreased phosphorylation of p70S6K and 4E-BP1/2 and inhibition of the canonical Wnt pathway with subsequent decrease of HB cell growth. This effect was dependent of high baseline Wnt activity either by mutational status of β-catenin or extrinsic Wnt activation. Wnt inhibition seemed to be strengthened by disruption of the FOXM1-β-catenin complex. Furthermore, treatment of HB cells with aprepitant led to reduced expression of (liver) stemness markers (AFP, CD13, SOX2, NANOG, and OCT4) and SFA when grown under cancer stem cell conditions. Taken together, we show for the first time that targeting the SP/NK1R signaling cascade inhibits canonical Wnt signaling in HB cells. These findings reveal important insight into the molecular mechanisms of the SP/NK1R complex as a critical component in a model of pediatric liver cancer and may support the development of novel therapeutic interventions for HB and other Wnt-activated cancers.


Generalized papular and sclerodermoid eruption: scleromyxedema.
Scleromyxedema (SM) is a rare chronic progressive and highly intractable cutaneous disease with unknown etiology, affecting both genders equally between 30 and 50 years. The disease is characterized with mucin deposits in the skin and/or other organs. In fact it is a clinicopathological subset of lichen myxedematosus (LM) according to a new classification. Sclerodermiform plaques and lichenoid papules are characteristic cutaneous lesions. An elevation of IgG λ (lambda) chain exists in most cases and extracutaneous involvement occurring with variable systemic findings is also detected. Generalized form is quite difficult to treat and may even be fatal. Herein, we present a male patient with typical features of generalized papular and sclerodermoid LM variety and with benign outcome by isotretinoin.


Predicting COVID-19 in China Using Hybrid AI Model.
The coronavirus disease 2019 (COVID-19) breaking out in late December 2019 is gradually being controlled in China, but it is still spreading rapidly in many other countries and regions worldwide. It is urgent to conduct prediction research on the development and spread of the epidemic. In this article, a hybrid artificial-intelligence (AI) model is proposed for COVID-19 prediction. First, as traditional epidemic models treat all individuals with coronavirus as having the same infection rate, an improved susceptible-infected (ISI) model is proposed to estimate the variety of the infection rates for analyzing the transmission laws and development trend. Second, considering the effects of prevention and control measures and the increase of the public's prevention awareness, the natural language processing (NLP) module and the long short-term memory (LSTM) network are embedded into the ISI model to build the hybrid AI model for COVID-19 prediction. The experimental results on the epidemic data of several typical provinces and cities in China show that individuals with coronavirus have a higher infection rate within the third to eighth days after they were infected, which is more in line with the actual transmission laws of the epidemic. Moreover, compared with the traditional epidemic models, the proposed hybrid AI model can significantly reduce the errors of the prediction results and obtain the mean absolute percentage errors (MAPEs) with 0.52%, 0.38%, 0.05%, and 0.86% for the next six days in Wuhan, Beijing, Shanghai, and countrywide, respectively.


Short-course prophylaxis against tuberculosis in HIV-infected persons. A decision and cost-effectiveness analysis.
Isoniazid prophylaxis for 12 months effectively prevents tuberculosis in HIV-infected persons and may decrease the incidence of other HIV-related disease and mortality. Recent clinical trials have found that some short-course regimens also effectively prevent tuberculosis. To compare the benefits, risks, and cost-effectiveness of isoniazid prophylaxis and short-course prophylaxis regimens. Decision and cost-effectiveness analysis. United States. Hypothetical patients who are HIV-infected and have CD4 counts of 200 cells/mm3 or less and positive results on tuberculin skin tests. Isoniazid prophylaxis lasting 12 months and six short-course prophylaxis regimens of isoniazid, rifampin, and pyrazinamide alone or in combination. 5-year survival rate, life expectancy, lifetime incidence of tuberculosis, and cost per quality-adjusted life-year saved. Compared with no prophylaxis, the 12-month isoniazid regimen increased 5-year survival rates by 9% and life expectancy by 8.7 months, decreased incidence of tuberculosis by 27%, and saved 4 medical care dollars for every 1 spent on prophylaxis. Regimens of isoniazid for 6 months, isoniazid and rifampin for 3 months, and rifampin and pyrazinamide for 2 months had similar results: 6.2- to 8.6-month increases in life expectancy, 19% to 26% reductions in incidence of tuberculosis, and 1 to 7 medical care dollars saved for every 1 spent on prophylaxis. A 3-month regimen of isoniazid, rifampin, and pyrazinamide resulted in fewer clinical benefits and was the only regimen tested that did not save medical care dollars. Prophylaxis decreases the incidence of tuberculosis and increases life expectancy for HIV-infected patients. Some regimens save medical care dollars, and some short-course regimens have clinical and economic benefits similar to those of the 12-month isoniazid regimen. Short-course prophylaxis is a reasonable alternative to the 12-month isoniazid regimen.


Early lethality, functional NF-kappaB activation, and increased sensitivity to TNF-induced cell death in TRAF2-deficient mice.
TRAF2 is an intracellular signal-transducing protein recruited to the TNFR1 and TNFR2 receptors following TNF stimulation. To investigate the physiological role of TRAF2, we generated TRAF2-deficient mice. traf2-/- mice appeared normal at birth but became progressively runted and died prematurely. Atrophy of the thymus and spleen and depletion of B cell precursors also were observed. Thymocytes and other hematopoietic progenitors were highly sensitive to TNF-induced cell death and serum TNF levels were elevated in these TRAF2-deficient animals. Examination of traf2-/- cells revealed a severe reduction in TNF-mediated JNK/SAPK activation but a mild effect on NF-kappaB activation. These results suggest that TRAF2-independent pathways of NF-kappaB activation exist and that TRAF2 is required for an NF-kappaB-independent signal that protects against TNF-induced apoptosis.


The ecotoxicology and chemistry of manufactured nanoparticles.
The emerging literature on the ecotoxicity of nanoparticles and nanomaterials is summarised, then the fundamental physico-chemistry that governs particle behaviour is explained in an ecotoxicological context. Techniques for measuring nanoparticles in various biological and chemical matrices are also outlined. The emerging ecotoxicological literature shows toxic effects on fish and invertebrates, often at low mg l(-1) concentrations of nanoparticles. However, data on bacteria, plants, and terrestrial species are particularly lacking at present. Initial data suggest that at least some manufactured nanoparticles may interact with other contaminants, influencing their ecotoxicity. Particle behaviour is influenced by particle size, shape, surface charge, and the presence of other materials in the environment. Nanoparticles tend to aggregate in hard water and seawater, and are greatly influenced by the specific type of organic matter or other natural particles (colloids) present in freshwater. The state of dispersion will alter ecotoxicity, but many abiotic factors that influence this, such as pH, salinity, and the presence of organic matter remain to be systematically investigated as part of ecotoxicological studies. Concentrations of manufactured nanoparticles have rarely been measured in the environment to date. Various techniques are available to characterise nanoparticles for exposure and dosimetry, although each of these methods has advantages and disadvantages for the ecotoxicologist. We conclude with a consideration of implications for environmental risk assessment of manufactured nanoparticles.


Field enhancement of MoS2: visualization of the enhancement and effect of the number of layers.
Two-dimensional transition metal dichalcogenides (2D TMDCs) are layered semiconductor materials with unique electronic and optical properties. In the family of 2D TMDCs, molybdenum disulphide (MoS2) is a promising material for next-generation optoelectrical devices due to its high mobility and characteristic properties. The properties of 2D TMDCs, as well as device performances, can be further improved by a field enhancement effect. However, field enhancement has not been reported to date in the 2D TMDC family. Here, we show the field enhancement of MoS2 and its dependence on the number of layers (5-850 layers). Measurements of the fluorescence intensity of a dye solution, crystal violet, were used to visualize the enhancement factor (EF) for a MoS2 flake as a map. The EFs on the map were independently confirmed by x-y-z size measurements of the same MoS2 flake with an atomic force microscope. Furthermore, the obtained x-y-z sizes of the MoS2 flake were used for the finite-difference time-domain (FDTD) calculations to evaluate field enhancement. As a result, the MoS2 flake with a specific thickness (ca. 80 layers) gave the highest enhancement with EF = 100. Theoretical calculations based on the Mie scattering theory also confirmed the experimental EF mapping results, the dependence on the number of layers, and the component analysis of field enhancement. As another crucial point, large and small enhancement effects were attributed to the electric field and charge transfer effects, respectively, both of which depend on the number of layers. A transition region of these effects was indicated at around 300-400 layers.


A Personal Narrative on Living and Dealing with Psychiatric Symptoms after DBS Surgery.
Although deep brain stimulation (DBS) may result in dramatic motor improvement in people with Parkinson's disease (PD), it has been correlated with a number of postoperative psychiatric side effects. We report a case of a person with PD experiencing depression and hypomania following DBS surgery. We provide a detailed report of the patient's personal experiences dealing with and managing these psychiatric side effects for three years. Providing a personal narrative focusing on detailed patient subjective experiences complements reports that give insight into the short- and long-term effects of DBS on established psychiatric measures and neurologic activity. But, most importantly, such a qualitative approach provides prospective patients and clinicians with a broader ethical picture of real-life challenges faced and coping strategies employed by PD patients treated with DBS who are experiencing psychiatric adverse events. This case study reinforces the ethical need to disclose the potential risk of harm to prospective patients as well as the importance of establishing a multidisciplinary postoperative supportive group.


Levels and Correlates of Objectively Measured Sedentary Behavior in Young Children: SUNRISE Study Results from 19 Countries.
There is a paucity of global data on sedentary behavior during early childhood. The purpose of this study was to examine how device-measured sedentary behavior in young children differed across geographically, economically, and sociodemographically diverse populations, in an international sample. This multinational, cross-sectional study included data from 1071 children 3-5 yr old from 19 countries, collected between 2018 and 2020 (pre-COVID). Sedentary behavior was measured for three consecutive days using activPAL accelerometers. Sedentary time, sedentary fragmentation, and seated transport duration were calculated. Linear mixed models were used to examine the differences in sedentary behavior variables between sex, country-level income groups, urban/rural settings, and population density. Children spent 56% (7.4 h) of their waking time sedentary. The longest average bout duration was 81.1 ± 45.4 min, and an average of 61.1 ± 50.1 min·d-1 was spent in seated transport. Children from upper-middle-income and high-income countries spent a greater proportion of the day sedentary, accrued more sedentary bouts, had shorter breaks between sedentary bouts, and spent significantly more time in seated transport, compared with children from low-income and lower-middle-income countries. Sex and urban/rural residential setting were not associated with any outcomes. Higher population density was associated with several higher sedentary behavior measures. These data advance our understanding of young children's sedentary behavior patterns globally. Country income levels and population density appear to be stronger drivers of the observed differences, than sex or rural/urban residential setting.


A critical view of metabolic network adaptations.
There has been considerable recent interest in deciphering the adaptive properties underlying the structure and function of metabolic networks. Various features of metabolic networks such as the global topology, distribution of fluxes, and mutational robustness, have been proposed to have adaptive significance and hence reflect design principles. However, whether evolutionary processes alternative to direct selection on the trait under investigation also play a role is often ignored and the selection pressures maintaining a given metabolic trait often remain speculative. Some systems-level traits might simply arise as by-products of selection on other traits or even through random genetic drift. Here, we ask which systems-level aspects of metabolism are likely to have adaptive utility and which could be better explained as by-products of other evolutionary forces. We conclude that the global topological characteristics of metabolic networks and their mutational robustness are unlikely to be directly shaped by natural selection. Conversely, models of optimal design revealed that various aspects of individual pathways and the behavior of the whole network show signs of adaptations, even though the exact selective forces often remain elusive. Comparative and experimental approaches, which so far have been relatively rarely employed, could help to distinguish between alternative adaptive scenarios.


Host and geographic structure of endophytic and endolichenic fungi at a continental scale.
Endophytic and endolichenic fungi occur in healthy tissues of plants and lichens, respectively, playing potentially important roles in the ecology and evolution of their hosts. However, previous sampling has not comprehensively evaluated the biotic, biogeographic, and abiotic factors that structure their communities. Using molecular data we examined the diversity, composition, and distributions of 4154 endophytic and endolichenic Ascomycota cultured from replicate surveys of ca. 20 plant and lichen species in each of five North American sites (Madrean coniferous forest, Arizona; montane semideciduous forest, North Carolina; scrub forest, Florida; Beringian tundra and forest, western Alaska; subalpine tundra, eastern central Alaska). Endolichenic fungi were more abundant and diverse per host species than endophytes, but communities of endophytes were more diverse overall, reflecting high diversity in mosses and lycophytes. Endophytes of vascular plants were largely distinct from fungal communities that inhabit mosses and lichens. Fungi from closely related hosts from different regions were similar in higher taxonomy, but differed at shallow taxonomic levels. These differences reflected climate factors more strongly than geographic distance alone. Our study provides a first evaluation of endophytic and endolichenic fungal associations with their hosts at a continental scale. Both plants and lichens harbor abundant and diverse fungal communities whose incidence, diversity, and composition reflect the interplay of climatic patterns, geographic separation, host type, and host lineage. Although culture-free methods will inform future work, our study sets the stage for empirical assessments of ecological specificity, metabolic capability, and comparative genomics.


Lymph node fibroblastic reticular cells deposit fibrosis-associated collagen following organ transplantation.
Although the immune response within draining lymph nodes (DLNs) has been studied for decades, how their stromal compartment contributes to this process remains to be fully explored. Here, we show that donor mast cells were prominent activators of collagen I deposition by fibroblastic reticular cells (FRCs) in DLNs shortly following transplantation. Serial analysis of the DLN indicated that the LN stroma did not return to its baseline microarchitecture following organ rejection and that the DLN contained significant fibrosis following repetitive organ transplants. Using several FRC conditional-knockout mice, we show that induction of senescence in the FRCs of the DLN resulted in massive production of collagen I and a proinflammatory milieu within the DLN. Stimulation of herpes virus entry mediator (HVEM) on FRCs by its ligand LIGHT contributed chiefly to the induction of senescence in FRCs and overproduction of collagen I. Systemic administration of ex vivo-expanded FRCs to mice decreased DLN fibrosis and strengthened the effect of anti-CD40L in prolonging heart allograft survival. These data demonstrate that the transformation of FRCs into proinflammatory myofibroblasts is critically important for the maintenance of a proinflammatory milieu within a fibrotic DLN.


Direct observation of the external force mediated conformational dynamics of an IHF bound Holliday junction.
We have investigated the isomerization dynamics and plausible energy landscape of 4-way Holliday junctions (4WHJs) bound to integration host factor (IHF, a DNA binding protein), considering the effect of applied external force, by single-molecule FRET methods. A slowing down of the forward as well as the backward rates of the isomerization process of the protein bound 4WHJ has been observed under the influence of an external force, which indicates an imposed restriction on the conformational switching. This has also been reflected by an increase in rigidity, as observed from the increase in the single-molecule FRET (smFRET)-anisotropy values (0.270 ± 0.012 to 0.360 ± 0.008). The application of an external force has assisted the conformational transitions to share the unstacked open structure intermediate, with different rate-limiting steps and a huge induced variation in the energy landscape. Furthermore, the associated landscape of the 4WHJ is visualized in terms of rarely interconverting states embedded into the two isoforms by using nonlinear dynamics analysis, which shows that the chaoticity of the system increases at intermediate force (0.4 to 1.6 pN). The identification of chaos in our investigation provides useful information for a comprehensive explanation of the origin of the complex behavior of the system, which effectively helps us to perceive the dynamics of IHF bound 4WHJs under the influence of external force, and also demonstrates the applicability of nonlinear dynamics analysis in the field of biology.


[Impact of alcohol use disorder on mortality: are there age and gender differences?].
To estimate excess mortality in patients with an alcohol use disorder and to calculate the population-based impact attributable to this disorder, with emphasis on differences according to gender and age. We carried out a longitudinal study of a dynamic retrospective cohort in which 7,109 patients (76.5% men) aged 18-64 years who started medical day treatment between 1997 and 2006 were recruited. Deaths were monitored until the end of 2006 using a national mortality register. Standardized mortality ratios (SMR) by age and gender and population attributable risks (PAR) by age group were estimated. There were 557 deaths (83% were men). The total excess mortality was 8 times higher than in the general population aged 18-64 years, especially in the group aged 25-34 years old (SMR: 11.2 [95% CI: 7.8-16.0] in men and 24 [IC95%: 11.5-50.4] in women). Significant differences by gender were found in all the variables analyzed. In Barcelona, alcohol use disorder was estimated to cause 73 deaths per year, representing 3.3% of the total annual deaths in this city in persons aged 18-64 years. This percentage was substantially higher in the groups aged 25-34 years (PAR: 19.4% [95% CI: 19.2-19.6]) and 18-24 years (PAR: 11.1% [95% CI: 10.7-11.4]). Women and young men with an alcohol use disorder have a higher risk of early mortality. Early detection of problematic alcohol consumption and selective and well-indicated prevention programs should be improved.


Peppermint oil in irritable bowel syndrome.
In a literature search 16 clinical trials investigating 180-200 mg enteric-coated peppermint oil (PO) in irritable bowel syndrome (IBS) or recurrent abdominal pain in children (1 study) with 651 patients enrolled were identified. Nine out of 16 studies were randomized double blind cross over trials with (n = 5) or without (n = 4) run in and/or wash out periods, five had a randomized double blind parallel group design and two were open labeled studies. Placebo served in 12 and anticholinergics in three studies as comparator. Eight out of 12 placebo controlled studies show statistically significant effects in favor of PO. Average response rates in terms of "overall success" are 58% (range 39-79%) for PO and 29% (range 10-52%) for placebo. The three studies versus smooth muscle relaxants did not show differences between treatments hinting for equivalence of treatments. Adverse events reported were generally mild and transient, but very specific. PO caused the typical GI effects like heartburn and anal/perianal burning or discomfort sensations, whereas the anticholinergics caused dry mouth and blurred vision. Anticholinergics and 5HT3/4-ant/agonists do not offer superior improvement rates, placebo responses cover the range as in PO trials. Taking into account the currently available drug treatments for IBS PO (1-2 capsules t.i.d. over 24 weeks) may be the drug of first choice in IBS patients with non-serious constipation or diarrhea to alleviate general symptoms and to improve quality of life.


Changes in density of on-premises alcohol outlets and impact on violent crime, Atlanta, Georgia, 1997-2007.
Regulating alcohol outlet density is an evidence-based strategy for reducing excessive drinking. However, the effect of this strategy on violent crime has not been well characterized. A reduction in alcohol outlet density in the Buckhead neighborhood of Atlanta from 2003 through 2007 provided an opportunity to evaluate this effect. We conducted a community-based longitudinal study to evaluate the impact of changes in alcohol outlet density on violent crime in Buckhead compared with 2 other cluster areas in Atlanta (Midtown and Downtown) with high densities of alcohol outlets, from 1997 through 2002 (preintervention) to 2003 through 2007 (postintervention). The relationship between exposures to on-premises retail alcohol outlets and violent crime were assessed by using annual spatially defined indices at the census block level. Multilevel regression models were used to evaluate the relationship between changes in exposure to on-premises alcohol outlets and violent crime while controlling for potential census block-level confounders. A 3% relative reduction in alcohol outlet density in Buckhead from 1997-2002 to 2003-2007 was associated with a 2-fold greater reduction in exposure to violent crime than occurred in Midtown or Downtown, where exposure to on-premises retail alcohol outlets increased. The magnitude of the association between exposure to alcohol outlets and violent crime was 2 to 5 times greater in Buckhead than in either Midtown or Downtown during the postintervention period. A modest reduction in alcohol outlet density can substantially reduce exposure to violent crime in neighborhoods with high density of alcohol outlets. Routine monitoring of community exposure to alcohol outlets could also inform the regulation of alcohol outlet density, consistent with Guide to Community Preventive Services recommendations.


Stereotactic Body Radiotherapy (SBRT) Reirradiation for Recurrent Pancreas Cancer.
After adjuvant or definitive radiation for pancreas cancer, there are limited conventional treatment options for recurrent pancreas cancer. We explored the role of (Stereotactic Body Radiotherapy) SBRT for reirradiation of recurrent pancreas Cancer. This is a retrospective study of patients reirradiated with SBRT for recurrent pancreas cancer. All patients were deemed unresectable and treated with systemic therapy. Fiducial gold markers were used. CT simulation was performed with oral and IV contrast and patients were treated with respiratory motion tracking in the Cyberknife(TM) system. 30 patients (17 men and 13 women) with a median age of 67 years were included in the study. The median target volume was 41.29cc. The median prescription dose was 25Gy (24-36Gy) in a median of 5 fractions prescribed to a mean 78% isodose line. The median overall survival was 14 months. The 1 and 2 year local control was 78%. The worst toxicity included 3/30(10%) Grade III acute toxicity for pain, bleeding and vomiting. There was 2/30 (7%) Grade III long-term bowel obstructions. SBRT can be a useful and tolerable option for patients with recurrent pancreas cancer after prior radiation.


Experimental studies of Rio Grande virus in rodent hosts.
Rio Grande (RG) virus, a new member of the Phlebotomus fever serogroup, was inoculated into wild wood rats (Neotoma micropus) and laboratory-reared cotton rats (Sigmodon hispidus) to determine if these potential hosts could be experimentally infected. Nine of 14 (64%) wood rats became viremic, with titers of circulating virus ranging from 10(2.3) to 10(5.3) plaque-forming units (PFU)/ml and a geometric mean titer of 10(3.7) PFU/ml. Virus was not detected in urine specimens from inoculated wood rats but was found in a single saliva specimen. RG virus was detected in the blood of 1 of 12 (8%) cotton rats. Neutralizing (N) antibody developed in 8 of 9 inoculated wood rats which survived for 30 days postinoculation and in 11 of 12 cotton rats. N antibody was still detectable in 4 of 7 wood rats which survived for 1 year, and all 7 were resistant to rechallenge with the virus, as were 3 wood rats with naturally-acquired antibody. High mortality (36%) occurred in inoculated wood rats; whereas low mortality (8%) occurred in cotton rats. The specific cause of death of the rats was not determined. Modes of transmission of the virus in nature are discussed.


Exercise training differentially affects intrinsic excitability of autonomic and neuroendocrine neurons in the hypothalamic paraventricular nucleus.
Oxytocinergic and vasopressinergic brain stem projections have been shown to play an important role in mediating cardiovascular adjustments during exercise training (ET). The aim of the present work was to determine whether the intrinsic excitability of hypothalamic neurons giving rise to brain stem peptidergic projections is altered as a consequence of ET. Whole cell patch-clamp recordings were obtained from nucleus of the solitarii tract (NTS)-projecting paraventricular nucleus of the hypothalamus (PVN) neurons and from supraoptic nucleus (SON) and PVN magnocellular cells (MNCs), in hypothalamic slices obtained from sedentary (S) and ET rats. Our results indicate that intrinsic excitability of PVN neurons that innervate the NTS (PVN-NTS) is enhanced by ET, resulting in a more efficient input-output function (increase number of evoked actions potentials, steeper frequency/current relationships and slower decaying frequency/time relationships). Changes in input-output function were accompanied by smaller hyperpolarizing afterpotentials (HAPs) and afterhyperpolarizing potentials (AHPs), during and after trains of spikes, respectively. On the other hand, a decreased efficacy in the input-output function was observed in SON/PVN MNCs during ET. Altogether, our results indicate that ET differentially affects the intrinsic excitability of autonomic and neurosecretory SON and PVN neurons. Increased excitability in PVN-NTS neurons may contribute to enhanced release of OT and VP peptides in the dorsal brain stem, and cardiovascular fine-tuning during exercise training.


Risk Factors and Mortality for Atypical Presentation of COVID-19 Infection in Hospitalized Patients - Lessons From the Early Pandemic.
To assess the clinical epidemiology and outcomes of patients hospitalized with COVID-19 who did not experience fever and cough during the early pandemic. Retrospective cohort of all patients admitted during March 13, 2020 through May 13, 2020 with laboratory-confirmed COVID-19 to 3 tertiary-care hospitals. Patient-level data (demographic, clinical manifestations, comorbid illnesses, inpatient treatment) were analyzed. The main outcome variable was atypical presentation, defined as any hospitalized patient with COVID-19 infection who did not experience both fever and cough. We identified risk factors for atypical presentation on univariate and multivariate analyses and assessed 30-day mortality differences via survival analysis. Of 163 patients in the study, 39 (24%) were atypical. On univariate analysis, atypical cases were significantly more likely to be older, reside in a long-term-care facility (LTCF), and have underlying diabetes mellitus, stroke, or cardiac disease; present without dyspnea or myalgia, have lower C-reactive proteins (CRP) and higher beta-natriuretic peptides. They were less likely to receive intensive care unit care or specific COVID-19 treatments (P < .05). The incidence of acute respiratory failure was not significantly different between the groups. On logistic regression, atypical cases were significantly more likely to be LTCF residents (P = 0.003) and have a lower average CRP (P = 0.01). Atypical cases had significantly higher 30-day mortality (hazard ratio 3.4 [95% CI, 1.6 - 7.2], P = 0.002). During the first pandemic surge, COVID-19 patients without inflammatory signs and symptoms were more likely to be LTCF residents and had higher mortality. Timely recognition of these atypical presentations may have prevented spread and improved clinical outcomes.


Victimization experiences of lesbian, gay, and bisexual individuals: a meta-analysis.
This meta-analysis quantitatively compiled the results of studies from 1992 to 2009 to determine the prevalence and types of victimization experienced by lesbian, gay, and bisexual (LGB) individuals. Based on the results of three searches, 386 studies were retrieved and coded. Comparisons were made across all LGB individuals (138 studies), between LGB and heterosexual individuals (65 studies), and between LGB females and males (53 studies), with over 500,000 participants. Multiple types of victimization were coded, including discrimination, physical assault, and school victimization. Findings revealed that for LGB individuals, reports of victimization experiences were substantial (e.g., 55% experienced verbal harassment, and 41% experienced discrimination) and some types have increased since a 1992 review, while others have decreased. LGB individuals experienced greater rates of victimization than heterosexual individuals (range: d = .04-.58). LGB males experienced some types of victimization more than LGB females (e.g., weapon assault and being robbed) but, overall, the gender differences were small. It can be concluded that LGB individuals still experience a substantial amount of victimization. Implications for research methods are discussed, including recommendations for sampling and measurement of victimization. [Supplementary materials are available for this article. Go to the publisher's online edition of Journal of Sex Research for the following free supplemental resource(s): Supplementary Tables. These tables are referred to in the text of this article as "Table S1," "Table S2," etc.].


Cortical and Commissural Defects Upon HCF-1 Loss in Nkx2.1-Derived Embryonic Neurons and Glia.
Formation of the cerebral cortex and commissures involves a complex developmental process defined by multiple molecular mechanisms governing proliferation of neuronal and glial precursors, neuronal and glial migration, and patterning events. Failure in any of these processes can lead to malformations. Here, we study the role of HCF-1 in these processes. HCF-1 is a conserved metazoan transcriptional co-regulator long implicated in cell proliferation and more recently in human metabolic disorders and mental retardation. Loss of HCF-1 in a subset of ventral telencephalic Nkx2.1-positive progenitors leads to reduced numbers of GABAergic interneurons and glia, owing not to decreased proliferation but rather to increased apoptosis before cell migration. The loss of these cells leads to development of severe commissural and cortical defects in early postnatal mouse brains. These defects include mild and severe structural defects of the corpus callosum and anterior commissure, respectively, and increased folding of the cortex resembling polymicrogyria. Hence, in addition to its well-established role in cell proliferation, HCF-1 is important for organ development, here the brain.


Characterization of an autotransporter adhesin protein shared by Burkholderia mallei and Burkholderia pseudomallei.
Autotransporters form a large family of outer membrane proteins specifying diverse biological traits of Gram-negative bacteria. In this study, we report the identification and characterization of a novel autotransporter gene product of Burkholderia mallei (locus tag BMA1027 in strain ATCC 23344). Database searches identified the gene in at least seven B. mallei isolates and the encoded proteins were found to be 84% identical. Inactivation of the gene encoding the autotransporter in the genome of strain ATCC 23344 substantially reduces adherence to monolayers of HEp-2 laryngeal cells and A549 type II pneumocytes, as well as to cultures of normal human bronchial epithelium (NHBE). Consistent with these findings, expression of the autotransporter on the surface of recombinant E. coli bacteria increases adherence to these cell types by 5-7 fold. The gene specifying the autotransporter was identified in the genome of 29 B. pseudomallei isolates and disruption of the gene in strain DD503 reduced adherence to NHBE cultures by 61%. Unlike B. mallei, the mutation did not impair binding of B. pseudomallei to A549 or HEp-2 cells. Analysis of sera from mice infected via the aerosol route with B. mallei and B. pseudomallei revealed that animals inoculated with as few as 10 organisms produce antibodies against the autotransporter, therefore indicating expression in vivo. Our data demonstrate that we have identified an autotransporter protein common to the pathogenic species B. mallei and B. pseudomallei which mediates adherence to respiratory epithelial cells and is expressed in vivo during the course of aerosol infection.


Molecular Architecture of Contactin-associated Protein-like 2 (CNTNAP2) and Its Interaction with Contactin 2 (CNTN2).
Contactin-associated protein-like 2 (CNTNAP2) is a large multidomain neuronal adhesion molecule implicated in a number of neurological disorders, including epilepsy, schizophrenia, autism spectrum disorder, intellectual disability, and language delay. We reveal here by electron microscopy that the architecture of CNTNAP2 is composed of a large, medium, and small lobe that flex with respect to each other. Using epitope labeling and fragments, we assign the F58C, L1, and L2 domains to the large lobe, the FBG and L3 domains to the middle lobe, and the L4 domain to the small lobe of the CNTNAP2 molecular envelope. Our data reveal that CNTNAP2 has a very different architecture compared with neurexin 1α, a fellow member of the neurexin superfamily and a prototype, suggesting that CNTNAP2 uses a different strategy to integrate into the synaptic protein network. We show that the ectodomains of CNTNAP2 and contactin 2 (CNTN2) bind directly and specifically, with low nanomolar affinity. We show further that mutations in CNTNAP2 implicated in autism spectrum disorder are not segregated but are distributed over the whole ectodomain. The molecular shape and dimensions of CNTNAP2 place constraints on how CNTNAP2 integrates in the cleft of axo-glial and neuronal contact sites and how it functions as an organizing and adhesive molecule.


Species diversity and biological invasions: relating local process to community pattern.
In a California riparian system, the most diverse natural assemblages are the most invaded by exotic plants. A direct in situ manipulation of local diversity and a seed addition experiment showed that these patterns emerge despite the intrinsic negative effects of diversity on invasions. The results suggest that species loss at small scales may reduce invasion resistance. At community-wide scales, the overwhelming effects of ecological factors spatially covarying with diversity, such as propagule supply, make the most diverse communities most likely to be invaded.


Phase behavior of the quantum Lennard-Jones solid.
The Lennard-Jones (LJ) potential is perhaps one of the most widely used models for the interaction of uncharged particles, such as noble gas solids. The phase diagram of the classical LJ solid is known to exhibit transitions between hcp and fcc phases. However, the phase behavior of the quantum LJ solid remains unknown. Thermodynamic integration based on path integral molecular dynamics (PIMD) and lattice dynamics calculations are used to study the phase stability of the hcp and fcc LJ solids. The hcp phase is shown to be stabilized by quantum effects in PIMD, while fcc is shown to be favored by lattice dynamics, which suggests a possible re-entrant low pressure fcc phase for highly quantum systems. Implications for the phase stability of noble gas solids are discussed. For parameters equating to helium, the expansion due to zero-point vibrations is associated with quantum melting: neither crystal structure is stable at zero pressure.


Correlation between herpes simplex virus neutralizing antibody titers determined by ELVIS cell and traditional plaque reduction assays.
Preventive viral vaccine efficacy trials require large-scale sample analysis to quantitate immune responses and their correlation with infection outcomes. Traditional plaque reduction assays measure a functionally important form of humoral immunity, neutralizing antibody titer. These assays, however, are time-consuming and laborious. We previously developed a higher throughput assay of neutralizing antibody to herpes simplex viruses 1 and 2 (Blevins et al., PLOS ONE, 10(12), e0144738) using the enzyme-linked virus inducible system (ELVIS) cell line; this cell line produces β-galactosidase in response to HSV infection. Here, serum samples from recipients of an investigational vaccine in the Herpevac Trial for Women were used to compare the ELVIS cell assay with the lower throughput, traditional plaque reduction assay. We demonstrate that neutralizing antibody titers to HSV-1 or HSV-2 determined using ELVIS cells positively correlate with neutralizing antibody titers determined by traditional plaque reduction assay, thus validating a higher throughput alternative for large-scale sample analysis.


The activated Notch1 signal pathway is associated with gastric cancer progression through cyclooxygenase-2.
Gastric carcinoma is one of the most common cancers and lethal malignancies worldwide. Thus far, the regulatory mechanisms of its aggressiveness are still poorly understood. To understand the pathogenesis and to develop new therapeutic strategies, it is essential to dissect the molecular mechanisms that regulate progression of gastric cancer. Herein, we sought to address whether Notch1 signal pathway is involved in the control of progression in gastric cancer. We found that expression of Notch ligand Jagged1 was correlated with aggressiveness of human gastric cancer. Patients with Jagged1 expression in gastric cancer tissues had a poor survival rate compared with those without Jagged1 expression. The Notch1 receptor intracellular domain (N1IC), the activated form of Notch1 receptor, promoted the colony-forming ability and xenografted tumor growth of human stomach adenocarcinoma SC-M1 cells. Migration and invasion abilities of SC-M1 cells were enhanced by N1IC. Furthermore, N1IC and C promoter-binding factor 1 (CBF1) bound to cyclooxygenase-2 (COX-2) promoter and elevated COX-2 expression in SC-M1 cells through a CBF1-dependent manner. The colony-forming, migration, and invasion abilities enhanced by N1IC were suppressed in SC-M1 cells after treatment with the COX-2 inhibitor NS-398 or knockdown of COX-2. These cellular processes inhibited by Notch1 knockdown were restored by prostaglandin E(2) or exogenous COX-2. Taken together, these results suggest that activation of Notch1 signal pathway promotes progression of gastric cancer, at least in part through COX-2.


Six new loci associated with body mass index highlight a neuronal influence on body weight regulation.
Common variants at only two loci, FTO and MC4R, have been reproducibly associated with body mass index (BMI) in humans. To identify additional loci, we conducted meta-analysis of 15 genome-wide association studies for BMI (n > 32,000) and followed up top signals in 14 additional cohorts (n > 59,000). We strongly confirm FTO and MC4R and identify six additional loci (P < 5 x 10(-8)): TMEM18, KCTD15, GNPDA2, SH2B1, MTCH2 and NEGR1 (where a 45-kb deletion polymorphism is a candidate causal variant). Several of the likely causal genes are highly expressed or known to act in the central nervous system (CNS), emphasizing, as in rare monogenic forms of obesity, the role of the CNS in predisposition to obesity.


Lessons learnt from early failure of a patient trial with a polymer-on-polymer resurfacing hip arthroplasty.
Background and purpose - Hip resurfacing (HR) is a treatment option promoted for hip arthritis in young and active patients. However, adverse reactions to metal are a concern and the search for non-metallic bearing options proceeds. We present the first clinical study performed in patients using a newly developed hydrophilic polymer-on-polymer hip resurfacing device. Patients and methods - After performing extensive hip simulator tests, biocompatibility testing and animal tests (ISO 14242-1,3; 10993-3,4,5,10,11), approval was obtained from the IRB committee to enroll 15 patients in the first clinical study in humans using this experimental polymer-on-polymer hip resurfacing device. All surgeries were done by 2 experienced hip resurfacing surgeons. Clinical scores and standard radiographs as well as routine MRIs were obtained at regular intervals. Results - The surgical technique proved feasible with successful implantation of the new device using PMMA cement fixation on both sides without complications. Postoperative imaging revealed a well-positioned and well-fixed polymer resurfacing hip arthroplasty in all 4 initial cases. All 4 patients were free of pain and had good function for the first 2 months. However, in all 4 cases early cup loosening occurred between 8 and 11 weeks after surgery, necessitating immediate closure of the study. All 4 patients had a reoperation and were revised to a conventional THA. Retrieval analyses confirmed early cup loosening at the implant-cement interface in all 4 cases. The femoral components remained well attached to the cement. The periprosthetic tissues showed only small amounts of polymeric wear debris and there was only a very mild inflammatory reaction to this. Interpretation - Early cup loosening mandated a premature arrest of this study. After additional laboratory testing this failure mode was found to be the result of a small, yet measurable contraction in the cup size after exposing these implants to biological fluid divalent ion fluctuations in vivo. Currently used preclinical tests had failed to detect this failure mechanism. Modification of the polymer is essential to overcome these problems and before the potential of a polymer-on-polymer resurfacing arthroplasty may be further evaluated in patients.


An Alpha Fetoprotein Producing Gastric Tumor with Yolk Sac, Hepatoid and Papillary Adenocarcinoma Components.
Alpha Fetoprotein (AFP) producing gastric carcinomas are very rare and have unique clinicopathological features and an extremely poor prognosis. Here, we report a case of AFP producing gastric carcinoma with three distinct histomorphologic patterns such as yolk sac like, hepatoid, tubular and papillary adenocarcinoma components. The uniqueness of this case is absence of metastases and associated findings such as fundic gland polyposis with varying degrees of dysplasia, gastric and duodenal well differentiated neuroendocrine tumour and rectal ganglioneuroma. The patient is symptom free as of two and half year's postoperative period. AFP producing gastric tumours although rare, need to be identified as it is known to carry poor prognosis.


Activation of the respiratory burst enzyme from human neutrophils in a cell-free system. Evidence for a soluble cofactor.
Activation of the respiratory burst in phagocytic cells, an important host defense process, is not yet well understood. We now report the development of a cell-free system for activation of NADPH oxidase, the respiratory burst enzyme, in human neutrophils. Activation was achieved by the addition of arachidonic acid to a postnuclear supernatant (500 g) from disrupted unstimulated cells (no arachidonate, 0.2; with arachidonate, 3.4 nmol superoxide anion/min per mg) and was dependent on both the concentration of arachidonate and on the amount of cellular material present. Activity stimulated by arachidonate appeared to be NADPH oxidase based on a Michaelis constant for NADPH of 32 microM and a pH optimum of 7.0-7.5. Separation of the 500-g supernatant by high speed centrifugation revealed a requirement for both soluble and particulate cofactors. Activation of NADPH oxidase by arachidonate did not occur in the high speed pellet fraction from unstimulated cells but could be restored by the addition of the high speed supernatant. In addition, priming of intact neutrophils with low concentrations of the chemoattractant N-formyl-methionyl-leucyl-phenylalanine or the tumor promoter phorbol myristate acetate replaced the soluble factor requirement for NADPH oxidase activation by arachidonate in the high speed pellet. This cell-free system can now be used to provide further insight into the biochemical basis of priming and the terminal mechanisms involved in the activation of NADPH oxidase.


The epidemiology of obesity: the size of the problem.
The epidemic of obesity took off from about 1980 and in almost all countries has been rising inexorably ever since. Only in 1997 did WHO accept that this was a major public health problem and, even then, there was no accepted method for monitoring the problem in children. It was soon evident, however, that the optimum population body mass index is about 21 and this is particularly true in Asia and Latin America where the populations are very prone to developing abdominal obesity, type 2 diabetes and hypertension. These features are now being increasingly linked to epigenetic programming of gene expression and body composition in utero and early childhood, both in terms of fat/lean tissue ratios and also in terms of organ size and metabolic pathway regulation. New Indian evidence suggests that insulin resistance at birth seems linked to low birth weight and a higher proportion of body fat with selective B12 deficiency and abnormalities of one carbon pool metabolism potentially responsible and affecting 75% of Indians and many populations in the developing world. Biologically there are also adaptive biological mechanisms which limit weight loss after weight gain and thereby in part account for the continuing epidemic despite the widespread desire to slim. Logically, the burden of disease induced by inappropriate diets and widespread physical inactivity can be addressed by increasing physical activity (PA), but simply advocating more leisure time activity is unrealistic. Substantial changes in urban planning and diet are needed to counter the removal of any every day need for PA and the decades of misdirected food policies which with free market forces have induced our current 'toxic environment'. Counteracting this requires unusual policy initiatives.