Biomechanical stability of a supra-acetabular pedicle screw internal fixation device (INFIX) vs external fixation and plates for vertically unstable pelvic fractures.
We have recently developed a subcutaneous anterior pelvic fixation technique (INFIX). This internal fixator permits patients to sit, roll over in bed and lie on their sides without the cumbersome external appliances or their complications. The purpose of this study was to evaluate the biomechanical stability of this novel supraacetabular pedicle screw internal fixation construct (INFIX) and compare it to standard internal fixation and external fixation techniques in a single stance pelvic fracture model. Nine synthetic pelves with a simulated anterior posterior compression type III injury were placed into three groups (External Fixator, INFIX and Internal Fixation). Displacement, total axial stiffness, and the stiffness at the pubic symphysis and SI joint were calculated. Displacement and stiffness were compared by ANOVA with a Bonferroni adjustment for multiple comparisons The mean displacement at the pubic symphysis was 20, 9 and 0.8 mm for external fixation, INFIX and internal fixation, respectively. Plate fixation was significantly stiffer than the INFIX and external Fixator (P = 0.01) at the symphysis pubis. The INFIX device was significantly stiffer than external fixation (P = 0.017) at the symphysis pubis. There was no significant difference in SI joint displacement between any of the groups. Anterior plate fixation is stiffer than both the INFIX and external fixation in single stance pelvic fracture model. The INFIX was stiffer than external fixation for both overall axial stiffness, and stiffness at the pubic symphysis. Combined with the presumed benefit of minimizing the complications associated with external fixation, the INFIX may be a more preferable option for temporary anterior pelvic fixation in situations where external fixation may have otherwise been used.


Gradients in the number of species at reef-seagrass ecotones explained by gradients in abundance.
Gradients in the composition and diversity (e.g. number of species) of faunal assemblages are common at ecotones between juxtaposed habitats. Patterns in the number of species, however, can be confounded by patterns in abundance of individuals, because more species tend to be found wherever there are more individuals. We tested whether proximity to reefs influenced patterns in the composition and diversity ('species density' = number of species per area and 'species richness' = number of species per number of individuals) of prosobranch gastropods in meadows of two seagrasses with different physiognomy: Posidonia and Amphibolis. A change in the species composition was observed from reef-seagrass edges towards the interiors of Amphibolis, but not in Posidonia meadows. Similarly, the abundance of gastropods and species density was higher at edges relative to interiors of Amphibolis meadows, but not in Posidonia meadows. However, species richness was not affected by proximity to reefs in either type of seagrass meadow. The higher number of species at the reef-Amphibolis edge was therefore a consequence of higher abundance, rather than species richness per se. These results suggest that patterns in the composition and diversity of fauna with proximity to adjacent habitats, and the underlying processes that they reflect, likely depend on the physiognomy of the habitat.


Kinetics of NO3- Influx in Spruce.
Influxes of 13NO3- across the root plasmalemma were measured in intact seedlings of Picea glauca (Moench) Voss. Three kinetically distinct uptake systems for NO3- were identified. In seedlings not previously exposed to external NO3-, a single Michaelis-Menten-type constitutive high-affinity transport system (CHATS) operated in a 2.5 to 500 [mu]M range of external NO3- [NO3-]o. The Vmax of this system was 0.1 [mu]mol g-1 h-1, and the Km was approximately 15 [mu]M. Following exposure to NO3- for 3 d, this CHATS activity was increased approximately 3-fold, with no change of Km. In addition, a NO3--inducible high-affinity system became apparent with a Km of approximately 100[mu]M. The combined Vmax for these discrete saturable components was 0.7 [mu]mol g-1 h-1. In both uninduced and induced plants a linear low-affinity system, additive to CHATS and an NO3--inducible high-affinity system, operated at [NO3-]o [greater than or equal to] 1 mM. The time taken to achieve maximal rates of uptake (full induction) was 2 d from 1.5 mM [NO3-]o and 3 d from 200 [mu]M [NO3-]o.


[Prevalence and associated factors of alcohol abuse and alcohol addiction].
The scope of this study is to estimate the prevalence of alcohol abuse and alcohol addiction and the respective associated factors in the urban population of a city of northeastern Brazil. It is a population-based cross-sectional study that investigated the consumption of alcohol of 270 people living in the urban area of Jequié, State of Bahia. Alcohol abuse, defined by a score of > 8 from responses to the Alcohol Use Disorders Identification Test (AUDIT). Alcohol addiction was identified by a score > 2 obtained in the application of the Cut down, Annoyed by criticism, Guilty and Eye-opener (CAGE). The analysis used the logistic regression model. The prevalence of alcohol abuse was 18.5%. For alcohol addiction it was 10.4%. After adjusted analysis, the groups with greater alcohol abuse were men (OR = 5.56) and young (OR = 5.41). Professing evangelical belief was inversely associated with alcohol abuse (OR = 0.04). An association between alcohol abuse was established in the young, males, and smoking, whereas the inverse association was observed with those professing evangelical religious beliefs.


Burden of diarrhea among children in Honduras, 2000-2004: estimates of the role of rotavirus.
To estimate the annual burden of diarrhea and of diarrhea that is associated with rotavirus (RV) in children who are treated at public clinics and hospitals in Honduras. Data were collected from computerized records of all children < 5 years old treated for diarrhea at clinics and hospitals operated by the Secretary of Health for the period of 2000 through 2004. A review of studies of RV in Honduras and neighboring countries provided estimates of detection rates of RV among children treated for acute diarrhea as outpatients or as inpatients. From these data, we estimated the annual number of cases of diarrhea and of rotavirus-related diarrhea in Honduras, the cumulative incidence of diarrhea and of rotavirus-related diarrhea for a child from birth to age 5 years, and the number of fatalities due to RV among children hospitalized for diarrhea. From 2000 through 2004, a mean of 222,000 clinic visits, 4,390 hospitalizations, and 162 in-hospital deaths due to diarrhea were recorded annually among children < 5 years of age in the public health facilities in Honduras. From our review of scientific literature on Honduras and neighboring countries, an estimated 30% of outpatients and 43% of inpatients who were treated for diarrhea would be expected to have RV. Consequently, we estimated that 66,600 outpatient visits, 1,888 hospitalizations, and 70 in-hospital deaths among children < 5 years in Honduras could be attributed to RV each year. Therefore, a child in the first five years of life has a respective risk for consultation, hospitalization, and in-hospital death of 1:1, 1:46, and 1:1,235 for diarrhea. For an episode associated with RV, the respective risks are 1:3, 1:106, and 1:2,857. These values likely underestimate the true burden of diarrhea in Honduras, since some 51% of children with acute diarrhea do not receive formal care for the illness, 70% do not receive oral rehydration solution, and 80% of diarrheal deaths occur outside of hospitals. Diarrhea is a major cause of illness among children < 5 years old in Honduras, and RV is likely the most common cause. Our preliminary estimates need to be refined so that health planners in Honduras can make decisions on the future use of rotavirus vaccines. A program of hospital-based surveillance for rotavirus in Honduras has been established to address this need.


The Technology of Processed Electroencephalogram Monitoring Devices for Assessment of Depth of Anesthesia.
Commercial brain function monitors for depth of anesthesia have been available for more than 2 decades; there are currently more than 10 devices on the market. Advances in this field are evidenced by updated versions of existing monitors, development of new monitors, and increasing research unveiling the mechanisms of anesthesia on the brain. Electroencephalography signal processing forms an integral part of the technology supporting the brain function monitors for derivation of a depth-of-anesthesia index. This article aims to provide a better understanding of the technology and functionality behind these monitors. This review will highlight the general design principles of these devices and the crucial stages in electroencephalography signal processing and classification, with a focus on the key mathematical techniques used in algorithm development for final derivation of the index representing anesthetic state. We will briefly discuss the advantages and limitations of this technology in the clinical setting as a tool in our repertoire used for optimizing individualized patient care. Also included is a table describing 10 available commercial depth-of-anesthesia monitors.


Genetic control of malaria parasite transmission: threshold levels for infection in an avian model system.
Genetic strategies for controlling malaria transmission based on engineering pathogen resistance in Anopheles mosquitoes are being tested in a number of animal models. A key component is the effector molecule and the efficiency with which it reduces parasite transmission. Single-chain antibodies (scFvs) that bind the circumsporozoite protein of the avian parasite, Plasmodium gallinaceum, can reduce mean intensities of sporozoite infection of salivary glands by two to four orders of magnitude in transgenic Aedes aegypti. Significantly, mosquitoes with as few as 20 sporozoites in their salivary glands are infectious for a vertebrate host, Gallus gallus. Although scFvs hold promise as effector molecules, they will have to reduce mean intensities of infection to zero to prevent parasite transmission and disease. We conclude that similar endpoints must be reached with human pathogens if we are to expect an effect on disease transmission.


The impact of hypothermia on outcomes in massively transfused patients.
Hypothermia is associated with poor outcomes after injury. The relationship between hypothermia during contemporary large volume resuscitation and blood product consumption is unknown. We evaluated this association, and the predictive value of hypothermia on mortality. Patients predicted to receive massive transfusion at 12 level 1 trauma centers were randomized in the Pragmatic Randomized Optimal Platelet and Plasma Ratios (PROPPR) trial and were grouped into those who were hypothermic (<36°C) or normothermic (36-38.5°C) within the first 6 hours of emergency department arrival. The impact of hypothermia or normothermia on the volume of blood product required during the first 24 hours was determined via negative binomial regression, adjusting for treatment arm, injury severity score, mechanism, demographics, pre-emergency department fluid volume, blood administered before becoming hypothermic, pulse and systolic blood pressure on arrival, and the time exposed to hypothermic or normothermic temperatures. Of 680 patients, 590 had a temperature measured during the first 6 hours in hospital, and 399 experienced hypothermia. The mean number of red blood cell (RBC) units given to all patients in the first 24 hours of admission was 8.8 (95% confidence interval [CI], 7.9-9.6). In multivariable analysis, every 1°C decrease in temperature below 36.0°C was associated with a 10% increase (incidence rate ratio, 0.90; 95% CI, 0.89-0.92; p < 0.00) in consumption of RBCs during the first 24 hours of admission. There was no association between RBC administration and a temperature above 36°C. Hypothermia on arrival was an independent predictor of mortality, with an adjusted odds ratio of 2.7 (95% CI, 1.7-4.5; p < 0.00) for 24-hour mortality and 1.8 (95% CI, 1.3-2.4; p < 0.00) for 30-day mortality. Hypothermia is associated with increase in blood product consumption and mortality. These findings support the maintenance of normothermia in trauma patients and suggest that further investigation on the impact of cooling or rewarming during massive transfusion is warranted. Prognostic, level III.


Does Physical Activity Mediate the Associations Between Local-Area Descriptive Norms, Built Environment Walkability, and Glycosylated Hemoglobin?
Associations between local-area residential features and glycosylated hemoglobin (HbA1c) may be mediated by individual-level health behaviors. Such indirect effects have rarely been tested. This study assessed whether individual-level self-reported physical activity mediated the influence of local-area descriptive norms and objectively expressed walkability on 10-year change in HbA1c. HbA1c was assessed three times for adults in a 10-year population-based biomedical cohort (n = 4056). Local-area norms specific to each participant were calculated, aggregating responses from a separate statewide surveillance survey for 1600 m road-network buffers centered on participant addresses (local prevalence of overweight/obesity (body mass index ≥25 kg/m²) and physical inactivity (<150 min/week)). Separate latent growth models estimated direct and indirect (through physical activity) effects of local-area exposures on change in HbA1c, accounting for spatial clustering and covariates (individual-level age, sex, smoking status, marital status, employment and education, and area-level median household income). HbA1c worsened over time. Local-area norms directly and indirectly predicted worsening HbA1c trajectories. Walkability was directly and indirectly protective of worsening HbA1c. Local-area descriptive norms and walkability influence cardiometabolic risk trajectory through individual-level physical activity. Efforts to reduce population cardiometabolic risk should consider the extent of local-area unhealthful behavioral norms and walkability in tailoring strategies to improve physical activity.


Internalization of Bordetella pertussis adenylate cyclase-haemolysin into endocytic vesicles contributes to macrophage cytotoxicity.
Bordetella pertussis adenylate cyclase-haemolysin is a critical virulence factor in the murine model of intranasal infection, where it is required for several pathological effects, including macrophage apoptosis. Based on biochemical and immunological properties, it was proposed that the toxin was delivered directly to the cytoplasm of eukaryotic cells without trafficking through the endocytic pathway. In the present study, we analysed the cellular distribution of the adenylate cyclase-haemolysin during intoxication of macrophages. We showed that, shortly after its initial binding to the plasma membrane of macrophages, the toxin gains access to intracellular compartments that become progressively positive for the endosomal marker transferrin, but not for the lysosomal membrane protein CD107a/Lamp1. Importantly, the vesicular trafficking of the adenylate cyclase-haemolysin appears to be required for its ability to induce macrophage death. Inhibitors of actin polymerization and of macropinocytosis, as well as depletion of plasma membrane cholesterol and disruption of the Golgi network, reduce the toxin's ability to kill macrophages. Altogether, these results suggest that internalization of the adenylate cyclase-haemolysin into endocytic vesicles, at least partly through macropinocytosis, contributes to cytotoxicity.


The Arabidopsis trithorax-like factor ATX1 functions in dehydration stress responses via ABA-dependent and ABA-independent pathways.
Emerging evidence suggests that the molecular mechanisms driving the responses of plants to environmental stresses are associated with specific chromatin modifications. Here, we demonstrate that the Arabidopsis trithorax-like factor ATX1, which trimethylates histone H3 at lysine 4 (H3K4me3), is involved in dehydration stress signaling in both abscisic acid (ABA)-dependent and ABA-independent pathways. The loss of function of ATX1 results in decreased germination rates, larger stomatal apertures, more rapid transpiration and decreased tolerance to dehydration stress in atx1 plants. This deficiency is caused in part by reduced ABA biosynthesis in atx1 plants resulting from decreased transcript levels from NCED3, which encodes a key enzyme controlling ABA production. Dehydration stress increased ATX1 binding to NCED3, and ATX1 was required for the increased levels of NCED3 transcripts and nucleosomal H3K4me3 that occurred during dehydration stress. Mechanistically, ATX1 affected the quantity of RNA polymerase II bound to NCED3. By upregulating NCED3 transcription and ABA production, ATX1 influenced ABA-regulated pathways and genes. ATX1 also affected the expression of ABA-independent genes, implicating ATX1 in diverse dehydration stress-response mechanisms in Arabidopsis.


Migrating curlews on schedule: departure and arrival patterns of a long-distance migrant depend on time and breeding location rather than on wind conditions.
Departure decisions in long-distance migratory bird species may depend on favourable weather conditions and beneficial resources at the destination location, overarched by genetic triggers. However, few studies have tried to validate the significance of these three concepts simultaneously, and long-term, high-resolution tagging datasets recording individual movements across consecutive years are scarce. We used such a dataset to explore intraspecific and intra-individual variabilities in departure and arrival decisions from/to wintering grounds in relation to these three different concepts in bird migration. We equipped 23 curlews (Numenius arquata) wintering in the Wadden Sea with Global Positioning System data loggers to record their spatio-temporal patterns of departure from and arrival at their wintering site, and the first part of their spring migration. We obtained data for 42 migrations over 6 years, with 12 individuals performing repeat migrations in consecutive years. Day of year of departure and arrival was related to 38 meteorological and bird-related predictors using the least absolute shrinkage and selection operator (LASSO) to identify drivers of departure and arrival decisions. Curlews migrated almost exclusively to Arctic and sub-Arctic Russia for breeding. They left their wintering site mainly during the evening from mid- to late April and returned between the end of June and mid-July. There was no difference in departure times between the sexes. Weather parameters did not impact departure decisions; if departure days coincided with headwind conditions, the birds accounted for this by flying at higher altitudes of up to several kilometres. Curlews breeding further away in areas with late snowmelt departed later. Departures dates varied by only < 4 days in individual curlews tagged over consecutive years. These results suggest that the trigger for migration in this long-distance migrant is largely independent of weather conditions but is subject to resource availability in breeding areas. The high intra-individual repeatability of departure days among subsequent years and the lack of relationship to weather parameters suggest the importance of genetic triggers in prompting the start of migration. Further insights into the timing of migration in immatures and closely related birds might help to further unravel the genetic mechanisms triggering migration patterns.


The effect of patient, fracture and surgery on outcomes of high energy neck of femur fractures in patients aged 15-50.
High-energy femoral neck fractures in young patients can be devastating, with the risk of osteonecrosis, nonunion, malunion and lifelong morbidity. The aim of this study is to define the effects of patient, fracture and surgical factors on the outcome of high-energy femoral neck fractures in patients aged from 15 to 50 years. A retrospective review was conducted of high-energy femoral neck fractures in patients aged 15-50 managed surgically at a Level 1 Trauma Centre, using a prospectively recorded trauma database. Low energy trauma (including falls from <1 m), medical conditions adversely affecting bone density, and pathological fractures were excluded. A clinical and radiological review was performed. The primary outcome measures were the development of osteonecrosis or nonunion leading to total hip arthroplasty (THA). Secondary outcome measures included osteotomy or other surgical procedures, quality of reduction and malunion. 32 patients meeting the inclusion criteria were identified between January 2008 and July 2015. The mean follow-up was 58.5 months (range 980-3,048 days). 3 patients (9.4%) required THA. No other surgical procedures were performed. None of the 29 other patients developed radiologically apparent osteonecrosis. Fracture type, displacement, anatomical reduction and fixation type were not statistically significant risk factors affecting these outcomes. For all patients, an average of 8% loss of femoral neck height and 10% femoral neck offset were seen. At a mean 4.9-year follow-up, the incidence of high-energy femoral neck fractures leading to THA was 9.4%, as a consequence of osteonecrosis or nonunion. Malunion was common.


[Evaluating tobacco control policy in Latin American countries during the era of the Framework Convention on Tobacco Control].
The Framework Convention on Tobacco Control (FCTC) aims to coordinate tobacco control policies around the world that reduce tobacco consumption. The FCTC's recommended policies are likely to be effective in low- and middle-income countries. Nevertheless, policy evaluation studies are needed to determine policy impact and potential synergies across policies. The International Tobacco Control Policy Evaluation Project (ITC) is an international collaboration to assess the psychosocial and behavioral impact of the FCTC's policies among adult smokers in nine countries. The ITC evaluation framework utilizes multiple country controls, a longitudinal design, and a theory-driven conceptual model to test hypotheses about the anticipated effects of given policies. ITC Project results generally confirm previous studies that form the evidence base for FCTC policy recommendations, in particular: the use of graphic warning labels; banning of "light" and "mild" descriptors; smoking bans; increasing tax and price; banning advertising; and using new cigarette product testing methods. Initial findings from the ITC Project suggest that Latin American countries could use similar methods to monitor and evaluate their own tobacco control policies while contributing to the evidence base for policy interventions in other countries.


De novo analysis of the transcriptome of Pratylenchus zeae to identify transcripts for proteins required for structural integrity, sensation, locomotion and parasitism.
The root lesion nematode Pratylenchus zeae, a migratory endoparasite, is an economically important pest of major crop plants (e.g. cereals, sugarcane). It enters host roots, migrates through root tissues and feeds from cortical cells, and defends itself against biotic and abiotic stresses in the soil and in host tissues. We report de novo sequencing of the P. zeae transcriptome using 454 FLX, and the identification of putative transcripts encoding proteins required for movement, response to stimuli, feeding and parasitism. Sequencing generated 347,443 good quality reads which were assembled into 10,163 contigs and 139,104 singletons: 65% of contigs and 28% of singletons matched sequences of free-living and parasitic nematodes. Three-quarters of the annotated transcripts were common to reference nematodes, mainly representing genes encoding proteins for structural integrity and fundamental biochemical processes. Over 15,000 transcripts were similar to Caenorhabditis elegans genes encoding proteins with roles in mechanical and neural control of movement, responses to chemicals, mechanical and thermal stresses. Notably, 766 transcripts matched parasitism genes employed by both migratory and sedentary endoparasites in host interactions, three of which hybridized to the gland cell region, suggesting that they might be secreted. Conversely, transcripts for effectors reported to be involved in feeding site formation by sedentary endoparasites were conspicuously absent. Transcripts similar to those encoding some secretory-excretory products at the host interface of Brugia malayi, the secretome of Meloidogyne incognita and products of gland cells of Heterodera glycines were also identified. This P. zeae transcriptome provides new information for genome annotation and functional analysis of possible targets for control of pratylenchid nematodes.


Regulation of brain temperature in pigeons: effects of corneal convection.
The effect of direct ventilation of the eyes on cooling in the brain was investigated in domestic pigeons (Columba livia, mean mass 0.27 kg) with thermocouples chronically implanted in the hypothalamus and anterior eye chamber. During conductive heating in still air body-brain temperature difference (delta T) was 2.6 degrees C. During exclusive ventilation of ocular surfaces, with air flowing at about flight speed, delta T increased to 3.5 degrees C and returned to preventilation values on cessation of ventilation. When the eyes were sealed then ventilated, delta T was not different from that in still air. Administration of phenylephrine caused iridial vasoconstriction and a significant decrease in intraocular temperature, but no changes in brain temperature. This suggests that compensation may occur via other evaporating cranial surfaces. Our findings suggest that the eyes contribute to the control of brain temperature by dissipating heat. Blood cooled while flowing through the ocular vasculature apparently contributes to the venous flow through the ophthalmic rete, serving as a heat sink for arterial blood flowing to the brain.


Sensitivity of the rat liver microsomal oxidation of pyrazole to antibody raised against the ethanol-inducible rabbit liver cytochrome P-450 isozyme.
Pyrazole is oxidized to 4-hydroxypyrazole by rat liver microsomes in a cytochrome P-450-dependent reaction and this oxidation can be increased by prior treatment of rats with pyrazole, 4-methylpyrazole, or chronic ethanol feeding. The induction pattern suggests that pyrazole may be an effective substrate for oxidation by P-450 IIE.1. This P-450 isozyme is recognized by antibody (anti-3a IgG) raised against the ethanol-inducible P-450 in rabbits. Experiments were carried out to evaluate the ability of anti-3a IgG to inhibit pyrazole oxidation by microsomes from controls and from rats treated with inducers of P-450 IIE.1. Immunoblots with anti-3a IgG or with the anti-pyrazole P-450 IgG were identical and indicated increased staining of the pyrazole P-450 with microsomes from rats treated with pyrazole, 4-methylpyrazole, or ethanol, relative to saline controls; very little staining occurred with microsomes from pair-fed controls or phenobarbital-treated rats. Rates of pyrazole oxidation were highest with microsomes from rats treated with the inducers of P-450 IIE.1 and lowest with pair-fed controls or rats treated with phenobarbital. Anti-3a IgG produced about a 60% decrease of pyrazole oxidation in microsomes from rats treated with inducers of P-450 IIE.1 and about a 25% decrease with the saline controls; no inhibition was found with microsomes from the phenobarbital-treated rats. The anti-3a IgG-resistant rate of pyrazole oxidation was similar with all the microsomal preparations, and was not due to interaction of pyrazole with hydroxyl radicals.(ABSTRACT TRUNCATED AT 250 WORDS)


Enhancement of Rhodamine B Degradation by Ag Nanoclusters-Loaded g-C₃N₄ Nanosheets.
In this paper, silver (Ag) nanoclusters-loaded graphitic carbon nitride (g-C₃N₄) nanosheets are synthesized and their physical properties as well as photocatalytic activities are systematically investigated by different techniques. The existence of Ag atoms in the form of nanoclusters (NCs) rather than well-crystallized nanoparticles are evidenced by X-ray diffraction patterns, SEM images, and XPS spectra. The deposition of Ag nanoclusters on the surface of g-C₃N₄ nanosheets affect the crystal structure and slightly reduce the band gap energy of g-C₃N₄. The sharp decrease of photoluminescence intensity indicates that g-C₃N₄/Ag heterojunctions successfully prevent the recombination of photo-generated electrons and holes. The photocatalytic activities of as-synthesized photocatalysts are demonstrated through the degradation of rhodamine B (RhB) solutions under Xenon lamp irradiation. It is demonstrated that the photocatalytic activity depends strongly on the molar concentration of Ag⁺ in the starting solution. The g-C₃N₄/Ag heterojunctions prepared from 0.01 M of Ag⁺ starting solution exhibit the highest photocatalytic efficiency and allow 100% degradation of RhB after being exposed for 60 min under a Xenon lamp irradiation, which is four times faster than that of pure g-C₃N₄ nanosheets.


Trajectories of autism severity in children using standardized ADOS scores.
To plot longitudinal trajectories of autism spectrum disorder (ASD) severity from early childhood to early adolescence. In line with reported trajectories in toddlers, we hypothesize that a substantial minority of children will show marked changes in ASD severity over time, with "Improvers" demonstrating the highest mean baseline and rate of growth in verbal IQ (VIQ). Patients included 345 clinic referrals and research participants with best-estimate clinical diagnoses of ASD at 1 or more time points, and repeated Autism Diagnostic Observation Schedule (ADOS), VIQ, and nonverbal IQ scores. Standardized ADOS severity scores were applied to 1026 assessments collected longitudinally between the ages of 2 and 15 (VIQ at most recent assessment: mean = 58, SD = 35). Scores were fitted for latent severity trajectory classes with and without covariates. Adaptive behavior and VIQ trajectories over time were modeled within each of the best-fit latent classes. A 4-class model best represented the observed data. Over 80% of participants were assigned to persistent (stable) high or moderately severe classes; 2 small classes respectively increased or decreased in severity over time. Age, gender, race, and nonverbal IQ did not predict class membership; VIQ was a significant predictor. Baseline VIQ was highest in the improving and worsening classes; it increased at the greatest rate in the improving class. Adaptive behavior declined in all but the improving class, with consistent impairment in all classes. If replicated, identified trajectory classes of ADOS severity may contribute to clinical prognosis and to subtyping samples for neurobiological and genetic research.


Therapeutic effects of chrysin in a rat model of traumatic brain injury: A behavioral, biochemical, and histological study.
Oxidative stress and apoptosis have major roles in the progression of traumatic brain injury (TBI)-associated motor and cognitive deficits. The present study was aimed to elucidate the putative effects of chrysin, a natural flavonoid compound, against TBI-induced motor and cognitive dysfunctions and possible involved mechanisms. Chrysin (25, 50 or 100 mg/kg) was orally administered to rats starting immediately following TBI induction by Marmarou's weight-drop technique and continuously for 3 or 14 days. Neurological functions, motor coordination, learning and memory performances, histological changes, cell apoptosis, expression of pro- and anti-apoptotic proteins, and oxidative status were assayed at scheduled time points after experimental TBI. The results indicated that treatment with chrysin improved learning and memory disabilities in passive avoidance task, and ameliorated motor coordination impairment in rotarod test after TBI. These beneficial effects were accompanied by increased the concentrations of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione (GSH), decreased malondialdehyde (MDA) content, prevented neuronal loss, diminished apoptotic index, elevated the expression of anti-apoptotic Bcl-2 protein, and reduced the expression of pro-apoptotic Bax protein in the cerebral cortex and hippocampus tissues. Our findings suggest that both anti-oxidative and anti-apoptotic properties of chrysin (especially in the dose of 100 mg/kg) are possible mechanisms that improve cognitive/motor deficits and prevent neuronal cell death after TBI.


Microfluidics for 3D Cell and Tissue Cultures: Microfabricative and Ethical Aspects Updates.
The necessity to improve in vitro cell screening assays is becoming ever more important. Pharmaceutical companies, research laboratories and hospitals require technologies that help to speed up conventional screening and therapeutic procedures to produce more data in a short time in a realistic and reliable manner. The design of new solutions for test biomaterials and active molecules is one of the urgent problems of preclinical screening and the limited correlation between in vitro and in vivo data remains one of the major issues. The establishment of the most suitable in vitro model provides reduction in times, costs and, last but not least, in the number of animal experiments as recommended by the 3Rs (replace, reduce, refine) ethical guiding principles for testing involving animals. Although two-dimensional (2D) traditional cell screening assays are generally cheap and practical to manage, they have strong limitations, as cells, within the transition from the three-dimensional (3D) in vivo to the 2D in vitro growth conditions, do not properly mimic the real morphologies and physiology of their native tissues. In the study of human pathologies, especially, animal experiments provide data closer to what happens in the target organ or apparatus, but they imply slow and costly procedures and they generally do not fully accomplish the 3Rs recommendations, i.e., the amount of laboratory animals and the stress that they undergo must be minimized. Microfluidic devices seem to offer different advantages in relation to the mentioned issues. This review aims to describe the critical issues connected with the conventional cells culture and screening procedures, showing what happens in the in vivo physiological micro and nano environment also from a physical point of view. During the discussion, some microfluidic tools and their components are described to explain how these devices can circumvent the actual limitations described in the introduction.


Anisotropic forces from spatially constrained focal adhesions mediate contact guidance directed cell migration.
Directed migration by contact guidance is a poorly understood yet vital phenomenon, particularly for carcinoma cell invasion on aligned collagen fibres. We demonstrate that for single cells, aligned architectures providing contact guidance cues induce constrained focal adhesion maturation and associated F-actin alignment, consequently orchestrating anisotropic traction stresses that drive cell orientation and directional migration. Consistent with this understanding, relaxing spatial constraints to adhesion maturation either through reduction in substrate alignment density or reduction in adhesion size diminishes the contact guidance response. While such interactions allow single mesenchymal-like cells to spontaneously 'sense' and follow topographic alignment, intercellular interactions within epithelial clusters temper anisotropic cell-substratum forces, resulting in substantially lower directional response. Overall, these results point to the control of contact guidance by a balance of cell-substratum and cell-cell interactions, modulated by cell phenotype-specific cytoskeletal arrangements. Thus, our findings elucidate how phenotypically diverse cells perceive ECM alignment at the molecular level.


Efficacy of hypobromous acid as a hide-on carcass antimicrobial intervention.
Escherichia coli O157:H7 and Salmonella on cattle hides at slaughter are the main source of beef carcass contamination by these foodborne pathogens during processing. Hypobromous acid (HOBr) has been approved for various applications in meat processing, but the efficacy of HOBr as a hide antimicrobial has not been determined. In this study, the antimicrobial properties of HOBr were determined by spraying cattle hides at either of two concentrations, 220 or 500 ppm. Treatment of hides with 220 ppm of HOBr reduced the prevalence of E. coli O157:H7 on hides from 25.3 to 10.1% (P < 0.05) and reduced the prevalence of Salmonella from 28.3 to 7.1% (P < 0.05). Treatment of hides with 500 ppm of HOBr reduced (P < 0.05) the prevalence of E. coli O157:H7 on hides from 21.2 to 10.1% and the prevalence of Salmonella from 33.3 to 8.1%. The application of 220 ppm of HOBr reduced (P < 0.05) aerobic plate counts, total coliform counts, and E. coli counts on hides by 2.2 log CFU/ 100 cm(2). The use of 500 ppm of HOBr resulted in reductions (P < 0.05) of aerobic plate counts, total coliform counts, and E. coli counts by 3.3, 3.7, and 3.8 log CFU/100 cm(2), respectively, demonstrating that the use of higher concentrations of HOBr on hides resulted in additional antimicrobial activity. These results indicate that the adoption of a HOBr hide wash will reduce hide concentrations of spoilage bacteria and pathogen prevalence, resulting in a lower risk of carcass contamination.


A Developmental Learning Approach of Mobile Manipulator via Playing.
Inspired by infant development theories, a robotic developmental model combined with game elements is proposed in this paper. This model does not require the definition of specific developmental goals for the robot, but the developmental goals are implied in the goals of a series of game tasks. The games are characterized into a sequence of game modes based on the complexity of the game tasks from simple to complex, and the task complexity is determined by the applications of developmental constraints. Given a current mode, the robot switches to play in a more complicated game mode when it cannot find any new salient stimuli in the current mode. By doing so, the robot gradually achieves it developmental goals by playing different modes of games. In the experiment, the game was instantiated into a mobile robot with the playing task of picking up toys, and the game is designed with a simple game mode and a complex game mode. A developmental algorithm, "Lift-Constraint, Act and Saturate," is employed to drive the mobile robot move from the simple mode to the complex one. The experimental results show that the mobile manipulator is able to successfully learn the mobile grasping ability after playing simple and complex games, which is promising in developing robotic abilities to solve complex tasks using games.


Corroboration of drug abusers' self-reports through the use of multiple data sources.
Multiple data sources were used to evaluate the validity of 31 drug abusers' self-reports of recent drug use as well as related behaviors occurring 1 year before and 1 year immediately after admission to a drug-free therapeutic community. Interviews were conducted by individuals not associated with the program. At the initial interview subjects provided a breath test for alcohol and a urine sample to test for other recent drug use. Subjects also signed releases of information so that official records could be checked both before and after admission to treatment to document any hospitalizations, arrests, drug treatment facility stays, and driver's license suspensions or revocations. These multiple data checks showed that drug abusers' self-reports of recent alcohol and drug use and of arrests and hospitalizations over a 2-year period are sufficiently valid for use in treatment outcome research. When differences occurred, it was usually due to subjects reporting more events than appeared on the records.


Norepinephrine removal and release in the forearm of healthy subjects.
The relevance of local removal and release of norepinephrine (NE) for antecubital venous plasma NE concentration was studied in 22 healthy subjects. Arterial and venous plasma NE and forearm blood flow were measured during intra-arterial infusion of two doses of NE, intra-arterial NE infusion with two doses of sodium nitroprusside, intravenous infusion of NE with intra-arterial infusion of four doses of sodium nitroprusside, and lower body negative pressure of -20 mm Hg for 15 minutes. The venous plasma NE concentration-time curves during the infusions of the two doses of NE indicated first-order kinetics for forearm extraction: forearm NE extraction rate during the low dose infusion was 67 +/- 4.1% (SEM) and correlated with basal forearm blood flow (r = -0.64, p less than 0.03, n = 12). Local sodium nitroprusside-induced vasodilatation during the intra-arterial and intravenous NE infusions was accompanied by dose-dependent decreases in forearm extraction rates for NE and epinephrine. During lower body negative pressure, taking into account the high basal forearm extraction rate for NE, local and systemic release of NE was indicated by increases in arterial and venous plasma and the venous-arterial plasma NE concentration difference (p less than 0.05 for all). These data show that removal of NE from forearm circulation is a process with a high extraction ratio obeying first-order kinetics and that this extraction process inversely relates to forearm blood flow. Thus, antecubital venous plasma NE is likely to be derived mainly from local release and not from the arterial plasma NE input.


Dynamic expression of pemphigus and desmosomal antigens by cultured keratinocytes.
Dynamic expression of pemphigus antigens by cultured human and mouse keratinocytes was compared with that of desmosome-associated molecules and cellular markers relating to epidermal differentiation. Plakoglobin was detected in localized areas of keratinocyte sheets in low Ca2+ (0.15 mM) KGM medium. In minimum essential medium (MEM) containing 1.8 mM Ca2+, plakoglobin was expressed in the intercellular spaces (ICS) throughout the keratinocyte sheet. Desmoplakin I and II, which were present in the cytoplasm of keratinocytes in the low Ca2+ medium, moved to the cell surface after the medium was changed to MEM. Desmoglein 1 and pemphigus vulgaris (PV) antigens were observed in the ICS of both the monolayers and stratified areas in the MEM. Pemphigus foliaceus (PF) antigens, frequently together with desmoglein 1, involucrin and keratins specific for the upper layer of the epidermis, were expressed by stratified keratinocytes but not the cells in the monolayers. The Western blotting study of the cultured keratinocyte extract showed 160- and 130-kDa bands positive for desmoglein 1 antigens and a 130-kDa band stained with PV sera. These findings suggest that although desmoglein 1 molecules bear PF antigenic sites, their expression pattern by cultured keratinocytes is closely related to that of PV rather than PF antigens. The PF antigenic sites may be formed on desmoglein 1 during epidermal differentiation.


Changing EDSS Progression in Placebo Cohorts in Relapsing MS: A Systematic Review and Meta-Regression.
Recent systematic reviews of randomised controlled trials (RCTs) in relapsing multiple sclerosis (RMS) revealed a decrease in placebo annualized relapse rates (ARR) over the past two decades. Furthermore, regression to the mean effects were observed in ARR and MRI lesion counts. It is unclear whether disease progression measured by the expanded disability status scale (EDSS) exhibits similar features. A systematic review of RCTs in RMS was conducted extracting data on EDSS and baseline characteristics. The logarithmic odds of disease progression were modelled to investigate time trends. Random-effects models were used to account for between-study variability; all investigated models included trial duration as a predictor to correct for unequal study durations. Meta-regressions were conducted to assess the prognostic value of a number of study-level baseline variables. The systematic literature search identified 39 studies, including a total of 19,714 patients. The proportion of patients in placebo controls experiencing a disease progression decreased over the years (p<0.001). Meta-regression identified associated covariates including the size of the study and its duration that in part explained the time trend. Progression probabilities tended to be lower in the second year of a study compared to the first year with a reduction of 28% in progression odds from year 1 to year 2 (p = 0.017). EDSS disease progression exhibits similar behaviour over time as the ARR and point to changes in trial characteristics over the years. This needs to be considered in comparisons between historical and recent trials.


Infants Attribute Value± to the Goal-Directed Actions of Self-propelled Objects.
Motion is a fundamental source of information for basic human interpretations; it is basic to the fundamental concept of causality and, the present model argues, equally basic to the fundamental concept of intentionality. The model is based on two main assumptions: When an infant perceives an object (1) moving spontaneously and (2) displaying goaldirected action, it will interpret the object as intentional and assign to it the unique properties of the psychological domain. The key property tested was: Do infants attribute value to interactions between intentional objects using criteria specified by the model? We showed infants (average age 52 weeks) computer-generated animations of spontaneously moving "balls," using looking time in a standard habituation/dishabituation paradigm. In two positive interactions, one ball either "caressed" another, or "helped" it achieve its goal; whereas in two negative interactions, one ball either "hit" another, or "prevented" it from achieving its goal. In keeping with predictions of the model, when transferred to a negative condition, infants who had been habituated on a positive condition showed greater dishabituation than those habituated on a negative condition. The results could not be easily explained by the similarity relations among the animations depicting the interactions. The results suggest that well before the age when the child can ascribe mental states or has a "theory of mind," it recognizes the goals of self-propelled objects and attributes value to the interactions between them.


Effects of ergonomic intervention in work with video display units.
This study evaluated the effect of an intensive ergonomic approach and education on workstation changes and musculoskeletal disorders among workers who used a video display unit (VDU). A randomized controlled design was used. The subjects (N=124) were allocated into three groups (intensive ergonomics, ergonomic education, reference) using stratified random sampling. The evaluation involved questionnaires, a diary of discomfort, measurements of workload, and an ergonomic rating of the workstations. The assessments were made 2 weeks before the intervention and after 2 and 10 months of follow-up. The intensive and training groups showed less musculoskeletal discomfort than the reference group after 2 months of follow-up. Positive effects on discomfort were seen primarily for the shoulder, neck, and upper back areas. No significant differences were found for the strain levels or prevalence of pain. After the intervention the ergonomic level was distinctly higher in the intensive ergonomic group than in the education or reference group. Both the intensive ergonomics approach and education in ergonomics help reduce discomfort in VDU work. In attempts to improve the physical ergonomics of VDU workstations, the best result will be achieved with cooperative planning in which both workers and practitioners are actively involved.


Ellagic acid inhibits proliferation and induces apoptosis in human glioblastoma cells.
To investigate the anticancer activity of ellagic acid (EA) in U251 human glioblastoma cells and its possible molecular mechanism. The cells were treated with EA at various concentrations for different time periods. Cell viability and cell proliferation were detected by cell counting kit-8(CCK-8) assay and live/dead assay respectively. Cell apoptosis were measured with Annexin V-FITC/PI double staining method by flow cytometry and Mitochondrial membrane potential assay separately. Cell cycle was measured with PI staining method by flow cytometry. The expressions of Bcl-2, Survivin, XIAP, Caspase-3, Bax, JNK, p-JNK, ERK1/2, p-ERK1/2, p38, p-p38, DR4, DR5, CHOP and GRP78-related proteins were detected by western blot after EA treatment. Cell viability and proliferation of glioblastoma cells treated with EA were significantly lower than the control group. EA caused robust apoptosis of the glioblastoma cells compared to the control group. EA significantly decreased the proportion at G0/G1 phases of cell cycling accompanied by increased populations at S phase in U251 cell lines. And the expressions of anti-apoptotic proteins were dramatically down-regulated. Ellagic acid potentially up-regulated DR4, DR5 and MAP kinases (JNK, ERK1/2 and p38). EA also caused significant increase in the expressions of CHOP and GRP78. Our findings suggest that EA would be beneficial for the treatment of glioblastoma.


Mycotic aneurysm secondary to melioidosis in China: A series of eight cases and a review of literature.
Burkholderia pseudomallei is the causative agent of melioidosis, endemic in Southeast Asia and Northern Australia, and increasingly recognized in southern China, especially in Hainan Province. Mycotic aneurysm caused by B. pseudomallei is a rare but potentially severe illness with a high mortality rate. The clinical features of the mycotic aneurysm secondary to melioidosis have not been illustrated in China. Over a seven-year period (2013 to 2019), 159 patients with bacteremic melioidosis were retrospectively analyzed in Hainan province, China, of whom eight patients were confirmed to have mycotic aneurysm through the combination of imaging examination, pathologic examination and aneurysm tissue culture. We summarized these eight patients' clinical characteristics, demographical features, treatments and outcomes. The susceptibilities to five commonly-used antibiotics for these eight B. pseudomallei isolates were also determined by E-test strips. Furthermore, the mycotic aneurysm cases secondary to melioidosis retrieved from the literature were also reviewed. Of the eight cases, six had abdominal mycotic aneurysms, one had a left iliac aneurysm, and the other one had an infectious mesenteric aneurysm. They were aged from 48 to 69 years old, and had the underlying risk factors of diabetes mellitus (2 patients), long-term smoking (4 patients), hypertension (6 patients), and soil and water contact history (6 patients), respectively. The positive arterial aneurysm imaging was observed in all patients via computed tomography (CT) or angiography. Eight B. pseudomallei isolates collected from both blood and mycotic aneurysm tissues remained 100% susceptible to imipenem and ceftazidime. After surgery combined with antibiotic administration, six patients survived, with a mortality rate of 25%. In melioidosis endemic areas, the mycotic aneurysm secondary to melioidosis might be underdiagnosed, and increased awareness of predisposing risk factors and clinical features of the mycotic aneurysm is required. Following a positive B. pseudomallei blood culture, the diagnosis of mycotic aneurysm should be under consideration in those with abdominal pain and/or hypertension. Imaging by CT or angiography is indispensable for its timely diagnosis and management.


[Opinions of primary care managers on sources of influence on medical practice. Differences with physicians' opinions].
To determine the opinion of primary healthcare managers on the importance and legitimacy of different sources of influence in medical practice, and to compare the results with the opinions of physicians in healthcare teams. cross-sectional study. primary healthcare managers in the Spanish public health system (area managers, medical and nursing directors) and in the Andalusian health service (district director, nursing coordinators and epidemiology and program coordinators). The sample comprised the total population of 302. As dependent variables, a series of questions was designed to gather the interviewees' opinions on different strategies, institutions and/or collectives that exert some kind of influence on medical practice. The degree of importance of each factor was summarized into a set of 9 items. The subjects were asked to score each item from 1 (most important) to 9 (least important). To assess the legitimacy of these scores, 16 items were presented measured using a Likert-type 7-point scale (1: not at all legitimate; 7: very legitimate). A self-administered questionnaire was used, sent by mail. Non-parametric tests (Friedman and Kruskall-Wallis) were used for statistical analysis of the data. The response rate was 79.8%. Using the Friedman test for an ordinal 9-point scale, analysis of the mean ranges for each item revealed that the most important sources of influence for the primary healthcare managers interviewed were: the devising of management protocols by the doctors themselves; discussion with colleagues; feedback from patients, and attending training courses, and reading articles and scientific journals. The institutions or groups with the greatest legitimacy to influence medical practice were: users or citizens; internal audits; peers; scientific associations, and the managers themselves. The sources of influence considered to have the greatest importance and legitimacy in influencing medical practice concern the professional medical system (self-defined protocols, discussion with colleagues, etc.). Managers accept the use of business managerial tools as well as the influence of social actors to a greater extent than do physicians. This finding could indicate differences in the value systems between primary healthcare physicians and managers.


Antiviral regimen complexity index as an independent predictor of sustained virologic response in patients with chronic hepatitis C.
Hepatitis C virus (HCV) infection affects more than 170 million people worldwide, and one-third of them have human immunodeficiency virus (HIV) coinfection. Multiple studies have been conducted in order to identify the factors that may explain different responses to treatment among patients. However, the reasons why HIV-HCV coinfected patients have lower responses to treatment are not clear. In addition, no studies have evaluated the influence of the complexity of the therapeutic regimen for hepatitis C infection on clinical outcomes. To (a) investigate the influence of the antiviral regimen complexity in the sustained viral response (SVR) in patients with chronic hepatitis and (b) adapt a method of quantifying complexity of an antiretroviral regimen for patients infected with HCV. A single center, retrospective study was conducted in HCV and HIV-HCV coinfected patients. We selected patients treated with interferon alfa-2a plus ribavirin between January 2005 and December 2010. Patients with severe psychiatric disorders, those included in a clinical trial, and those known to be nonadherent to treatment were excluded. The dependent variable was the sustained virologic response and the independent variables were sex, age, race, stage fibrosis (F) ≥2, presence or absence of cirrhosis, low hepatitis C baseline viral load (defined as ≤800,000 IU), viral genotype, rapid virological response (RVR), serum gamma-glutamyltransferase (GGT) levels, ratio of alanine aminotransferase to aspartate aminotransferase (ALT/AST), serum cholesterol level, presence or absence of diabetes mellitus, and antiviral regimen complexity index. The latter variable included drugs for HCV and HIV infection, but no medication for other comorbidities. To evaluate the complexity of antiviral treatment we performed an adaptation of the system developed by Martin et al. (2007) in HIV patients. The factors determining the complexity of treatment were the number of medications, dosing schedules, administration methods, special instructions, and required preparations associated with antiviral regimens. Sample size was estimated by the Freeman equation. To determine the independent variables associated with SVR, we performed an univariate logistic regression and subsequently a multivariate analysis with those variables that demonstrated a statistically significant difference in the univariate analysis. A total of 156 patients was included (76% men, mean age 44 years) of whom 45% were HIV-HCV coinfected. 75% were genotypes 1 or 4. The univariate analysis variables-genotypes 2 and 3 (OR=3.10; CI [1.38-6.95]; P=0.006); HIV-HCV coinfection (OR=0.36; CI [0.19-0.69]); presence of cirrhosis (OR=0.27; CI [0.10-0.73]; P=0,01); F≥2 (OR=0.44; CI [0.23-0.84]; P=0.01); low baseline viral load (OR= 2.05; CI [1.01-4.17]; P=0.048); RVR (OR=17,60; CI [6.84-45.30]; P less than 0.001); complexity index (OR=0.71; CI [0.58-0.87]; P=0.001), showed statistically significant relationships with SVR. Complexity index (OR=0.67; CI [0.52-0.87]; P=0.002) and RVR (OR=20.04; CI [7.33-54.85]; P less than 0.001) were independent predictors of SVR in multivariate analysis. The reliability of the multivariate analysis was checked with the Hosmer and Lemeshow test (P=0.079). The medication regimen complexity may be a crucial factor to achieve therapeutic success when treating patients for hepatitis C. The adaptation of this index in patients with HCV provides an objective value of the antiviral regimen complexity and could help us to identify patients in clinical practice who require multidisciplinary attention. Simplification of the antiretroviral regimen might result in a greater response to treatment for hepatitis C.


Genetic predisposition to diabetic nephropathy. Evidence for a role of the angiotensin I--converting enzyme gene.
In search of genetic determinants of susceptibility to diabetic nephropathy, we examined the association between DNA sequence differences at the locus of angiotensin I-converting enzyme (ACE) and renal complications in 151 insulin-dependent diabetes mellitus (IDDM) patients with a diabetes duration of 16-21 years. This nested case-control study included 77 normoalbuminuric control subjects (albumin excretion rate < 30 micrograms/min) and 74 cases with evidence of nephropathy ranging from microalbuminuria to overt proteinuria. DNA from each of these patients was genotyped at the ACE locus by a three-allele restriction fragment-melting polymorphism (RFMP) (Dde I), which we described recently, and a two-allele insertion/deletion recognized as an Xba I restriction fragment-length polymorphism, which has been shown by other investigators to be associated with serum levels of ACE and with risk of myocardial infarction. The least common allele of the Dde I RFMP was significantly more frequent among cases with nephropathy than among normoalbuminuric control subjects (12.8 vs. 4.5%, P < 0.05). The deletion in the ACE gene was also more frequent in case than in control subjects (56.1 vs. 47.4%), but the difference was not statistically significant (P < 0.25) with this sample size. To determine the independence of these associations, the two polymorphisms were analyzed jointly to identify Xba I/Dde I haplotypes. As might be expected, carriers of the Xba I/Dde I '+ =' haplotype had a fourfold risk of developing diabetic nephropathy (odds ratio [OR] 4.0, 95% confidence interval [CI] 1.5-11.0). However, this did not explain all of the excess Xba I '+' allele among cases.(ABSTRACT TRUNCATED AT 250 WORDS)


A heat shock protein 70-based vaccine with enhanced immunogenicity for clinical use.
In previous studies, we have shown that heat shock protein 70-peptide complexes (HSP70.PCs) derived from the fusion of dendritic cells (DCs) to tumor cells (HSP70.PC-F) possess superior properties compared with HSP70.PCs from tumor cells. HSP70.PC-F are more effective in stimulation of DC maturation and induction of CTL that are able to provide protection of mice against challenge with tumor cells. To develop an improved formulation of HSP70.PC-based tumor vaccine for patient use, we extracted HSP70.PC-F from DCs fused to patient-derived ovarian cancer cells or established human breast cancer cells and examined their properties as tumor vaccines. HSP70.PC-F induced T cells that expressed higher levels of IFN-gamma and exhibited increased levels of killing of tumor cells, compared with those induced by HSP70.PC derived from tumor cells. Enhanced immunogenicity of HSP70.PC-F was associated with improved composition of the vaccine, including increased content of tumor Ags and their processed intermediates, and the detection of other heat shock proteins (HSPs) such as HSP90 and HSP110. The present study has therefore provided an alternative approach to preparation of HSP-based vaccines using DC/tumor fusion technology and gentle and rapid isolation of HSP peptide complexes.


Why medical students do not like to join rural health service? An exploratory study in India.
Inadequate, inequitable distribution of the medical workforce remains a challenge across the globe, and India is no exception. Odisha, a state in India faces a major shortage of doctors particularly in rural and remote areas. In order to address this challenge, it is essential to understand medical students' career plans, specialization preferences, choices of job location and sector, and views on working in rural and remote areas. This study explored the immediate and long-term career plans of final year medical students, their intended practice locations and underlying reasons for the choices. A cross-sectional survey was conducted in all the medical colleges (three government and three private) in the state of Odisha. Through the systematic sampling method, data were gathered from 390 final year students. A semi-structured questionnaire was administered to the students and data were analyzed using SPSS version 20. Of the 390 students, 290 (74.35%) were from a government college. The most preferred immediate career goal was postgraduation studies (45.9% of students in government medical schools and 54% in private). About 17% of government students and 9% of private students showed willingness to work in rural areas, in the long run. Nearly 44.5% mentioned opportunities for career growth, followed by the possibilities for higher education (26.8%) as major the factors for preferring an urban posting. Similarly, higher pay scales, better working conditions were major factors for preferring the private sector. Most of the students maintained that good housing, better salaries, and adequate facilities at the workplace would attract more students toward rural service. Since public funded medical students are not motivated to serve in rural settings, increasing the number of places or establishing new medical institutions may not be an effective solution to the issue. Approaches such as extended clinical apprenticeship in rural health facilities, long-term community engagement during medical studentship could be considered.


Transforming the learning outcomes of anaesthesiology training into entrustable professional activities: A Delphi study.
True competency-based medical education should produce graduates meeting fixed standards of competence. Current postgraduate programmes are usually based on a pre-determined length of time in training making them poorly suited for an individual approach. The concept of entrustable professional activities (EPAs) enables a more flexible, personalised and less time-dependent approach to training programmes. An EPA is a unit of professional practice, to be entrusted to a trainee to execute without supervision once they demonstrate sufficient competence. As EPAs relate competencies to clinical practice, they structure training and assessment more logically according to the way clinicians actually work. A first step in building an EPA-based curriculum is to identify the core EPAs of the profession. The aim of this study was to identify EPAs for postgraduate training in anaesthesiology and to provide an example of how an existing curriculum can be transformed into an EPA-based curriculum. A modified Delphi method was used as a consensus approach applying three Delphi rounds. Postgraduate specialty training in anaesthesiology in the Netherlands. All programme directors in anaesthesiology in the Netherlands except for a single programme director who was involved as a researcher in this study and could not participate. Agreement among participants on a list of EPAs. Agreement was specified as a consensus rate of more than 80%. In this study, 27 programme directors (69% overall response rate) reached consensus on a set of 45 EPAs that describe a curriculum in anaesthesiology for the Netherlands. This study is a first step toward a more contemporary curriculum in competency-based postgraduate anaesthesiology training.


Temporal dynamics in animal community assembly during post-logging succession in boreal forest.
Species assemblages can result from deterministic processes, such as niche differentiation and interspecific interactions, and from stochastic processes, such as random colonisation and extinction events. Although changes in animal communities following disturbances have been widely examined, few studies have investigated the mechanisms structuring communities during ecological succession. We assessed the impact of logging on small mammal and beetle assemblages in landscapes dominated by old-growth boreal forests. Our objectives were to 1) characterize variations in communities during the first 66 years of post-harvest forest succession, 2) determine if there are non-random patterns of species co-occurrence (i.e., deterministic processes), and if there are, 3) establish whether non-random co-occurrences are best explained by habitat attributes or by interspecific interactions. We captured small mammals and beetles along a gradient of forest succession (5-66 years) and in old-growth forest, and characterized key vegetation attributes. First, we tested whether community compositions in clear-cut stands became similar to those in natural stands after 66 years. We then used null models, which were either unconstrained or constrained by habitat attributes, to address the last two objectives and distinguish effects of vegetation attributes from interspecific interactions on community assembly. We showed that beetle assemblages differed in stands 21-30 years post-harvest compared to old-growth forests. In contrast, harvesting did not influence the composition of small mammal communities. Overall, our results suggest that community assembly during forest succession is driven by both stochastic and deterministic processes, the latter being linked to interspecific interactions more strongly than to vegetation attributes.


Group B streptococcus neonatal invasive infections, France 2007-2012.
Streptococcus agalactiae (group B streptococcus (GBS)) is the leading cause of invasive infections among newborns in industrialized countries, with two described syndromes: early-onset disease (EOD) and late-onset disease (LOD). Since the introduction in many countries of intrapartum antibioprophylaxis (IAP), the incidence of EOD has dramatically decreased, whereas that of LOD remains unchanged. We describe the clinical and bacteriological characteristics of 438 GBS neonatal invasive infections notified to the French National Reference Centre for Streptococci in France from 2007 to 2012. Clinical data were retrieved from hospitalization reports or questionnaires. Capsular type, assignment to the hypervirulent clonal complex (CC)17 and antibiotic susceptibility profiles were determined. One hundred and seventy-four (39.7%) and 264 (60.3%) isolates were responsible for EOD, including death in utero, and LOD, respectively. EOD was associated with bacteraemia (n = 103, 61%) and LOD with meningitis (n = 145, 55%). EOD was mainly due to capsular polysaccharide (CPS) III isolates (n = 99, 57%) and CPS Ia isolates (n = 40, 23%), and CPS III isolates were responsible for 80% (n = 211) of LOD cases. CC17 accounted for 80% (n = 121) of CPS III isolates responsible for meningitis (n = 151; total cases of meningitis, 188). Bad outcome risk factors were low gestational age and low birthweight. LOD represents almost 60% of cases of neonatal GBS disease in France and other countries in which IAP has been implemented. This observation reinforces the need to develop new prevention strategies targeting CC17, which is predominant in GBS neonatal infections.


Physician-patient communication about prescription medication nonadherence: a 50-state study of America's seniors.
Understanding and improving the quality of medication management is particularly important in the context of the Medicare prescription drug benefit that took effect last January 2006. To determine the prevalence of physician-patient dialogue about medication cost and medication adherence among elderly adults nationwide. Cross-sectional survey. National stratified random sample of community-dwelling Medicare beneficiaries aged 65 and older. Rates of physician-patient dialogue about nonadherence and cost-related medication switching. Forty-one percent of seniors reported taking five or more prescription medications, and more than half has 2 or more prescribing physicians. Thirty-two percent overall and 24% of those with 3 or more chronic conditions reported not having talked with their doctor about all their different medicines in the last 12 months. Of seniors reporting skipping doses or stopping a medication because of side effects or perceived nonefficacy, 27% had not talked with a physician about it. Of those reporting cost-related nonadherence, 39% had not talked with a physician about it. Thirty-eight percent of those with cost-related nonadherence reported switching to a lower priced drug, and in a multivariable model, having had a discussion about drug cost was significantly associated with this switch (odds ratio [OR] 5.04, 95% confidence interval [CI] 4.28-5.93, P < .001). We show that there is a communication gap between seniors and their physicians around prescription medications. This communication problem is an important quality and safety issue, and takes on added salience as physicians and patients confront new challenges associated with coverage under new Medicare prescription drug plans. Meeting these challenges will require that more attention be devoted to medication management during all clinical encounters.


Growth hormone use in pediatric growth hormone deficiency and other pediatric growth disorders.
The diagnosis and management of growth disorders in children, particularly disorders that respond to therapy with growth hormone (GH), raise challenging clinical and economic issues. Several such issues are presented in the following article in which Dr. Ron Rosenfeld examines the evaluation and diagnosis of the child with short stature; Dr. David B. Allen discusses the anabolic and metabolic indications for GH treatment in children; Dr. Margaret H. MacGillivray reviews GH dosing, height outcomes, and follow up; and Dr. Craig Alter presents the payer's perspective on the diagnosis and treatment of pediatric GH deficiency. In addressing the use of GH in other pediatric populations, Dr. Paul Saenger focuses on Turner syndrome, Dr. Henry Anhalt on chronic renal insufficiency of childhood, and Dr. Ray Hintz on idiopathic short stature. Dr. Harvey P. Katz presents one managed care organization's policy and implementation plan that is used to guide decisions regarding coverage for GH treatment.


Iron Ladies - How Desiccated Asexual Rotifer Adineta vaga Deal With X-Rays and Heavy Ions?
Space exposure experiments from the last 15 years have unexpectedly shown that several terrestrial organisms, including some multi-cellular species, are able to survive in open space without protection. The robustness of bdelloid rotifers suggests that these tiny creatures can possibly be added to the still restricted list of animals that can deal with the exposure to harsh condition of space. Bdelloids are one of the smallest animals on Earth. Living all over the world, mostly in semi-terrestrial environments, they appear to be extremely stress tolerant. Their desiccation tolerance at any stage of their life cycle is known to confer tolerance to a variety of stresses including high doses of radiation and freezing. In addition, they constitute a major scandal in evolutionary biology due to the putative absence of sexual reproduction for at least 60 million years. Adineta vaga, with its unique characteristics and a draft genome available, was selected by ESA (European Space Agency) as a model system to study extreme resistance of organisms exposed to space environment. In this manuscript, we documented the resistance of desiccated A. vaga individuals exposed to increasing doses of X-ray, protons and Fe ions. Consequences of exposure to different sources of radiation were investigated in regard to the cellular type including somatic (survival assay) and germinal cells (fertility assay). Then, the capacity of A. vaga individuals to repair DNA DSB induced by different source of radiation was investigated. Bdelloid rotifers represent a promising model in order to investigate damage induced by high or low LET radiation. The possibility of exposure both on hydrated or desiccated specimens may help to decipher contribution of direct and indirect radiation damage on biological processes. Results achieved through this study consolidate our knowledge about the radioresistance of A. vaga and improve our capacity to compare extreme resistance against radiation among living organisms including metazoan.


Association between Diet-Quality Scores, Adiposity, Total Cholesterol and Markers of Nutritional Status in European Adults: Findings from the Food4Me Study.
Diet-quality scores (DQS), which are developed across the globe, are used to define adherence to specific eating patterns and have been associated with risk of coronary heart disease and type-II diabetes. We explored the association between five diet-quality scores (Healthy Eating Index, HEI; Alternate Healthy Eating Index, AHEI; MedDietScore, MDS; PREDIMED Mediterranean Diet Score, P-MDS; Dutch Healthy Diet-Index, DHDI) and markers of metabolic health (anthropometry, objective physical activity levels (PAL), and dried blood spot total cholesterol (TC), total carotenoids, and omega-3 index) in the Food4Me cohort, using regression analysis. Dietary intake was assessed using a validated Food Frequency Questionnaire. Participants (n = 1480) were adults recruited from seven European Union (EU) countries. Overall, women had higher HEI and AHEI than men (p < 0.05), and scores varied significantly between countries. For all DQS, higher scores were associated with lower body mass index, lower waist-to-height ratio and waist circumference, and higher total carotenoids and omega-3-index (p trends < 0.05). Higher HEI, AHEI, DHDI, and P-MDS scores were associated with increased daily PAL, moderate and vigorous activity, and reduced sedentary behaviour (p trend < 0.05). We observed no association between DQS and TC. To conclude, higher DQS, which reflect better dietary patterns, were associated with markers of better nutritional status and metabolic health.


A comprehensive scoping review of ability and disability in ADHD using the International Classification of Functioning, Disability and Health-Children and Youth Version (ICF-CY).
This is the first in a series of four empirical investigations to develop International Classification of Functioning, Disability and Health (ICF) Core Sets for Attention Deficit Hyperactivity Disorder (ADHD). The objective here was to use a comprehensive scoping review approach to identify the concepts of functional ability and disability used in the scientific ADHD literature and link these to the nomenclature of the ICF-CY. Systematic searches were conducted using Medline/PubMed, PsycINFO, ERIC and Cinahl, to extract the relevant concepts of functional ability and disability from the identified outcome studies of ADHD. These concepts were then linked to ICF-CY by two independent researchers using a standardized linking procedure. Data from identified studies were analysed until saturation of ICF-CY categories was reached. Eighty studies were included in the final analysis. Concepts contained in these studies were linked to 128 ICF-CY categories. Of these categories, 68 were considered to be particularly relevant to ADHD (i.e., identified in at least 5 % of the studies). Of these, 32 were related to Activities and participation, 31 were related to Body functions, and five were related to environmental factors. The five most frequently identified categories were school education (53 %), energy and drive functions (50 %), psychomotor functions (50 %), attention functions (49 %), and emotional functions (45 %). The broad variety of ICF-CY categories identified in this study underlines the necessity to consider ability and disability in ADHD across all dimensions of life, for which the ICF-CY provides a valuable and universally applicable framework. These results, in combination with three additional preparatory studies (expert survey, focus groups, clinical study), will provide a scientific basis to define the ICF Core Sets for ADHD for multi-purpose use in basic and applied research, and every day clinical practice.


The potential of fatty acid isotopes to trace trophic transfer in aquatic food-webs.
Compound-specific isotope analyses (CSIA) of fatty acids (FA) constitute a promising tool for tracing energy flows in food-webs. However, past applications of FA-specific carbon isotope analyses have been restricted to a relatively coarse food-source separation and mainly quantified dietary contributions from different habitats. Our aim was to evaluate the potential of FA-CSIA to provide high-resolution data on within-system energy flows using algae and zooplankton as model organisms. First, we investigated the power of FA-CSIA to distinguish among four different algae groups, namely cyanobacteria, chlorophytes, haptophytes and diatoms. We found substantial within-group variation but also demonstrated that δ13C of several FA (e.g. 18:3ω3 or 18:4ω3) differed among taxa, resulting in group-specific isotopic fingerprints. Second, we assessed changes in FA isotope ratios with trophic transfer. Isotope fractionation was highly variable in daphnids and rotifers exposed to different food sources. Only δ13C of nutritionally valuable poly-unsaturated FA remained relatively constant, highlighting their potential as dietary tracers. The variability in fractionation was partly driven by the identity of food sources. Such systematic effects likely reflect the impact of dietary quality on consumers' metabolism and suggest that FA isotopes could be useful nutritional indicators in the field. Overall, our results reveal that the variability of FA isotope ratios provides a substantial challenge, but that FA-CSIA nevertheless have several promising applications in food-web ecology. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.


Imaging the Injured Lung: Mechanisms of Action and Clinical Use.
Acute respiratory distress syndrome (ARDS) consists of acute hypoxemic respiratory failure characterized by massive and heterogeneously distributed loss of lung aeration caused by diffuse inflammation and edema present in interstitial and alveolar spaces. It is defined by consensus criteria, which include diffuse infiltrates on chest imaging-either plain radiography or computed tomography. This review will summarize how imaging sciences can inform modern respiratory management of ARDS and continue to increase the understanding of the acutely injured lung. This review also describes newer imaging methodologies that are likely to inform future clinical decision-making and potentially improve outcome. For each imaging modality, this review systematically describes the underlying principles, technology involved, measurements obtained, insights gained by the technique, emerging approaches, limitations, and future developments. Finally, integrated approaches are considered whereby multimodal imaging may impact management of ARDS.


Influence of saccadic adaptation on spatial localization: comparison of verbal and pointing reports.
Under conditions of short-term saccadic adaptation, stimuli presented long before saccadic onset are perceptually mislocalized in space. Here we study whether saccadic adaptation can also affect localization of objects by pointing. We measured localization performance during fixation and after normal saccades and adapted saccades, for a bar presented well before a saccadic eye movement, for both pointing and verbal localization, under open-loop conditions generated by a transient dark period about 300 ms after the presentation of the bar. During fixation and normal saccade, localization performance for verbal report was veridical, while for pointing there was an overestimation of the target eccentricity respect to gaze, in agreement with the idea of separate representations of space for action and perception. During saccadic adaptation, there was a significant shift of both pointing and verbal report localization in the direction of adaptation with similar spatial selectivity for both tasks. These results indicate that saccadic adaptation induces a similar re-calibration of the action map as well as of the perceptual map, suggesting a common site of operation in the transformation from eye-centered to gaze-centered coordinates.


Occupational risks for cutaneous melanoma among men in Sweden.
A population-based linked-registry was used to evaluate incidence of malignant melanoma of the skin among Swedish men by industry and occupation. There were 3850 cutaneous melanoma cases identified in the 19-year follow-up of men employed in 1960. New associations were observed for men employed in the breweries and malt-processing industry and in shoe fabrication from leather and skins. Several findings supported associations previously reported in other countries, including an excess risk among workers in basic chemical production and the printing industry and among professional, technical, and white-collar workers. Risk overall was not increased among farmers, despite a significant excess of melanoma of the face, neck, and scalp. Although this linked registry analysis lacked information about specific agents, duration of employment, and occupational and recreational sun exposures, it did provide leads for new associations and confirmed previous ones. Nevertheless, because of these limitations, etiologic clues must be interpreted cautiously.


Patient-driven loop control for ambulation function restoration in a non-invasive functional electrical stimulation system.
In this study, a patient-driven loop control in a non-invasive functional electrical stimulation (FES) system was designed to restore ambulation function of patients with stroke with their residual capabilities. With this patient-driven loop control, patients use the electromyographic (EMG) signals from their voluntary controlled muscles in affected extremity to adjust stimulus parameters of the system. A special designed FES system generated electrical stimuli to excite the paralysed muscles through surface electrodes on the basis of the control command from the residual myoelectric signals. The EMG signals were also served as the trigger and the adjustment of stimulus parameters and thereby adding versatility of the FES system. One patient with hemiplegia was recruited to conduct clinical evaluation and treated by using the new closed-loop FES system. The experimental results showed that hemiplegic could successfully control the system to restore their lost ambulation functions with the strategy of patient-driven loop control. It is revealed that the mean velocity, cadence, stride length, active ankle motion range and functional ambulation category have improved significantly from 0.22 + or - 0.17 m/s, 37.3 + or - 15.5 steps/min, 0.32 + or - 0.11 m, 10 degrees , level 2 to 0.46 + or - 0.23 m/s, 58.2 + or - 19.1 steps/min, 0.73 + or - 0.22 m, 35 degrees , level 4 respectively for the patient. A paired t-test indicated that differences in the EMG of the tibialis anterior and the gastrocnemius muscles between patient's disabled (affected-side) foot and normal (unaffected-side) foot are not significant (p > 0.05) after 12 weeks of training. According to the experiment results, this patient-driven loop control can be beneficial for patient with hemiplegia to restore their ambulation functions such as dorsi-flexion and plantar-flexion. The control strategy of this study has the potential to be employed not only in the FES system but also in other assistive devices.


SNAP, Young Children's Health, and Family Food Security and Healthcare Access.
The Supplemental Nutrition Assistance Program (SNAP) is the largest nutrition assistance program in the U.S. This study's objective was to examine the associations between SNAP participation and young children's health and development, caregiver health, and family economic hardships. Cross-sectional data from 2006 to 2016 were analyzed in 2017 for families with children aged <3 years in 5 cities. Generalized estimating equations and logistic regression were used to evaluate the associations of SNAP participation with child and caregiver health outcomes and food insecurity, forgone health care, and health cost sacrifices. Nonparticipants that were likely to be eligible for SNAP were compared with SNAP participants and analyses adjusted for covariates including Consumer Price Index for food to control for site-specific food prices. The adjusted odds of fair or poor child health status (AOR=0.92, 95% CI=0.86, 0.98), developmental risk (AOR=0.82, 95% CI=0.69, 0.96), underweight, and obesity in children were lower among SNAP participants than among nonparticipants. In addition, food insecurity in households and among children, and health cost sacrifices were lower among SNAP participants than among nonparticipants. Participation in SNAP is associated with reduced household and child food insecurity, lower odds of poor health and growth and developmental risk among infants and toddlers, and reduced hardships because of healthcare costs for their families. Improved SNAP participation and increased SNAP benefits that match the regional cost of food may be effective preventive health strategies for promoting the well-being of families with young children.


Mars: occurrence of liquid water.
In the absence of juvenile liquid water, condensation of water vapor to ice and subsequent melting of ice are the only means of producing liquid water on the martian surface. However, the evaporation rate is so high that the available heat sources cannot melt pure ice. Liquid water is therefore limited to concentrated solutions of strongly deliquescent salts.


Predictors of functional decline in elderly patients undergoing transcatheter aortic valve implantation (TAVI).
Aims This study aimed to assess functional course in elderly patients undergoing transcatheter aortic valve implantation (TAVI) and to find predictors of functional decline. Methods and results In this prospective cohort, functional course was assessed in patients ≥70 years using basic activities of daily living (BADL) before and 6 months after TAVI. Baseline EuroSCORE, STS score, and a frailty index (based on assessment of cognition, mobility, nutrition, instrumental and basic activities of daily living) were evaluated to predict functional decline (deterioration in BADL) using logistic regression models. Functional decline was observed in 22 (20.8%) of 106 surviving patients. EuroSCORE (OR per 10% increase 1.18, 95% CI: 0.83-1.68, P = 0.35) and STS score (OR per 5% increase 1.64, 95% CI: 0.87-3.09, P = 0.13) weakly predicted functional decline. In contrast, the frailty index strongly predicted functional decline in univariable (OR per 1 point increase 1.57, 95% CI: 1.20-2.05, P = 0.001) and bivariable analyses (OR: 1.56, 95% CI: 1.20-2.04, P = 0.001 controlled for EuroSCORE; OR: 1.53, 95% CI: 1.17-2.02, P = 0.002 controlled for STS score). Overall predictive performance was best for the frailty index [Nagelkerke's R(2) (NR(2)) 0.135] and low for the EuroSCORE (NR(2) 0.015) and STS score (NR(2) 0.034). In univariable analyses, all components of the frailty index contributed to the prediction of functional decline. Conclusion Over a 6-month period, functional status worsened only in a minority of patients surviving TAVI. The frailty index, but not established risk scores, was predictive of functional decline. Refinement of this index might help to identify patients who potentially benefit from additional geriatric interventions after TAVI.


A computational model for the analysis of lipoprotein distributions in the mouse: translating FPLC profiles to lipoprotein metabolism.
Disturbances of lipoprotein metabolism are recognized as indicators of cardiometabolic disease risk. Lipoprotein size and composition, measured in a lipoprotein profile, are considered to be disease risk markers. However, the measured profile is a collective result of complex metabolic interactions, which complicates the identification of changes in metabolism. In this study we aim to develop a method which quantitatively relates murine lipoprotein size, composition and concentration to the molecular mechanisms underlying lipoprotein metabolism. We introduce a computational framework which incorporates a novel kinetic model of murine lipoprotein metabolism. The model is applied to compute a distribution of plasma lipoproteins, which is then related to experimental lipoprotein profiles through the generation of an in silico lipoprotein profile. The model was first applied to profiles obtained from wild-type C57Bl/6J mice. The results provided insight into the interplay of lipoprotein production, remodelling and catabolism. Moreover, the concentration and metabolism of unmeasured lipoprotein components could be determined. The model was validated through the prediction of lipoprotein profiles of several transgenic mouse models commonly used in cardiovascular research. Finally, the framework was employed for longitudinal analysis of the profiles of C57Bl/6J mice following a pharmaceutical intervention with a liver X receptor (LXR) agonist. The multifaceted regulatory response to the administration of the compound is incompletely understood. The results explain the characteristic changes of the observed lipoprotein profile in terms of the underlying metabolic perturbation and resultant modifications of lipid fluxes in the body. The Murine Lipoprotein Profiler (MuLiP) presented here is thus a valuable tool to assess the metabolic origin of altered murine lipoprotein profiles and can be applied in preclinical research performed in mice for analysis of lipid fluxes and lipoprotein composition.


NspS, a predicted polyamine sensor, mediates activation of Vibrio cholerae biofilm formation by norspermidine.
Vibrio cholerae is both an environmental bacterium and a human intestinal pathogen. The attachment of bacteria to surfaces in biofilms is thought to be an important feature of the survival of this bacterium both in the environment and within the human host. Biofilm formation occurs when cell-surface and cell-cell contacts are formed to make a three-dimensional structure characterized by pillars of bacteria interspersed with water channels. In monosaccharide-rich conditions, the formation of the V. cholerae biofilm requires synthesis of the VPS exopolysaccharide. MbaA (locus VC0703), an integral membrane protein containing a periplasmic domain as well as cytoplasmic GGDEF and EAL domains, has been previously identified as a repressor of V. cholerae biofilm formation. In this work, we have studied the role of the protein NspS (locus VC0704) in V. cholerae biofilm development. This protein is homologous to PotD, a periplasmic spermidine-binding protein of Escherichia coli. We show that the deletion of nspS decreases biofilm development and transcription of exopolysaccharide synthesis genes. Furthermore, we demonstrate that the polyamine norspermidine activates V. cholerae biofilm formation in an MbaA- and NspS-dependent manner. Based on these results, we propose that the interaction of the norspermidine-NspS complex with the periplasmic portion of MbaA diminishes the ability of MbaA to inhibit V. cholerae biofilm formation. Norspermidine has been detected in bacteria, archaea, plants, and bivalves. We suggest that norspermidine serves as an intercellular signaling molecule that mediates the attachment of V. cholerae to the biotic surfaces presented by one or more of these organisms.


Heart transplantation is associated with an increased risk for pancreaticobiliary disease.
To determine the risk factors for and the incidence, morbidity, mortality, and outcome of pancreaticobiliary disease in patients who have had orthotopic heart transplantation. Retrospective case-control analysis. University hospital-based heart transplantation center. 20 case-patients with pancreaticobiliary disease and 40 controls; all patients received heart transplants between 1985 and 1994. Controls were matched to case-patients by time of transplantation and length of survival. Charts were reviewed and data were extracted using a structured data abstraction protocol. Risk factors assessed before transplantation were cause of heart disease, history of diabetes, reproductive history, and sex. Risk factors assessed at presentation of pancreaticobiliary disease were weight change after transplantation, alcohol use, use of medications, recent total parenteral nutrition, age at symptom onset, recent rejection episode, cyclosporine level, complete blood count, time between transplantation and onset of symptoms, body mass index, calcium level, liver function test results before and at symptom onset, and lipid profile. Pancreaticobiliary disease occurred in 20 of 255 transplant recipients (7.8%). The incidence of disease in these patients within 1 year after transplantation was 3.9% compared with an expected rate of 0.2% per year (P < 0.01). A decreased serum calcium level was the only risk factor found to differ significantly between the two groups (mean +/- SD, 2.19 +/- 0.17 mmol/L in case-patients and 2.31 +/- 0.14 mmol/L in controls; P = 0.005). The duration of hospitalization for the treatment of pancreaticobiliary disease was longer for patients who received transplants than for patients who did not receive transplants and were treated at Temple University Hospital during a similar period (17.1 days compared with 7.2 days; P < 0.001). However, the outcome was excellent in most patients. Pancreaticobiliary disease occurs 17.4 times (95% CI, 9.2 to 32.7 times) more frequently in patients receiving transplants than in the general population. It is a seldom recognized but apparently common complication of orthotopic heart transplantation that results in substantial morbidity and health care resource use. Further study is needed to ascertain why this condition occurs and how the risk for developing it can be reduced.


Establishment of subcellular fractionation techniques to monitor the intracellular fate of polymer therapeutics I. Differential centrifugation fractionation B16F10 cells and use to study the intracellular fate of HPMA copolymer - doxorubicin.
Polymer therapeutics are being designed for lysosomotropic, endosomotropic and transcellular drug delivery. Their appropriate intracellular routing is thus crucial for successful use. For example, polymer-anticancer drug conjugates susceptible to lysosomal enzyme degradation will never deliver their drug payload unless they encounter the appropriate activating enzymes. Many studies use confocal microscopy to monitor intracellular fate, but there is a pressing need for more quantitative methods able to define intracellular compartmentation over time. Only then will it be possible to optimise the next generation of polymer therapeutics for specific applications. The aim of this study was to establish a subcellular fractionation method for B16F10 murine melanoma cells and subsequently to use it to define the intracellular trafficking of N-(2-hydroxyproplylmethacrylamide) (HPMA) copolymer-bound doxorubicin (PK1). Free doxorubicin was used as a reference. The cell cracker method was used to achieve cell breakage and optimised to reproducibly achieve approximately 90% breakage efficiency. This ensured that subsequent subcellular fractionation experiments were representative for the whole cell population. To characterise the subcellular fractions obtained by differential centrifugation, DNA (nuclei), succinate dehydrogenase (mitochondria), N-acetyl-beta-glucosaminidase (lysosomes), alkaline phosphatase (plasma membrane) and lactate dehydrogenase (cytosol) were selected as markers and their assay was carefully validated. The relative specific activity (RSA) of the fractions obtained from B16F10 cells were: nuclei (2.2), mitochondria (4.1), lysosomes (3.7) and cytosol (2.5). When used to study the intracellular distribution at non-toxic concentrations of PK1 and doxorubicin, time-dependent accumulation of PK1 in lysosomes was evident and the expected nuclear localisation of free doxorubicin was seen. Live cell fluorescence microscopy and confocal co-localisation studies gave qualitative corroboration of these results, but by using this method, we were unable to accurately define organelle localisation. In conclusion, the B16F10 subcellular fractionation method developed here provides a useful tool to allow comparison of the intracellular trafficking of other polymer conjugates.