Mycobacteria release active membrane vesicles that modulate immune responses in a TLR2-dependent manner in mice.
Bacteria naturally release membrane vesicles (MVs) under a variety of growth environments. Their production is associated with virulence due to their capacity to concentrate toxins and immunomodulatory molecules. In this report, we show that the 2 medically important species of mycobacteria, Mycobacterium tuberculosis and Mycobacterium bovis bacille Calmette-Guérin, release MVs when growing in both liquid culture and within murine phagocytic cells in vitro and in vivo. We documented MV production in a variety of virulent and nonvirulent mycobacterial species, indicating that release of MVs is a property conserved among mycobacterial species. Extensive proteomic analysis revealed that only MVs from the virulent strains contained TLR2 lipoprotein agonists. The interaction of MVs with macrophages isolated from mice stimulated the release of cytokines and chemokines in a TLR2-dependent fashion, and infusion of MVs into mouse lungs elicited a florid inflammatory response in WT but not TLR2-deficient mice. When MVs were administered to mice before M. tuberculosis pulmonary infection, an accelerated local inflammatory response with increased bacterial replication was seen in the lungs and spleens. Our results provide strong evidence that actively released mycobacterial vesicles are a delivery mechanism for immunologically active molecules that contribute to mycobacterial virulence. These findings may open up new horizons for understanding the pathogenesis of tuberculosis and developing vaccines.


The impact of Nepal's national incentive programme to promote safe delivery in the district of Makwanpur.
Nepal's Safe Delivery Incentive Programme (SDIP) was introduced nationwide in 2005 with the aim of encouraging greater use of professional care at childbirth. It provided cash to women giving birth in a public health facility and an incentive to the health provider for each delivery attended, either at home or in the facility. We aimed to assess the impact of the programme on neonatal mortality and health care seeking behaviour at childbirth in one district of Nepal. Impacts were identified using an interrupted time series approach, applied to houSehold data. We estimated a model linking the level of each outcome at a point in time to the start of the programme, demographic controls, a vector of time variables and community-level fixed effects. The recipients of the cash transfer in the programme's first two years were disproportionately wealthier households, reflecting existing inequality in the use of government maternity services. In places with women's groups--where information about the policy was widely disseminated--the SDIP substantially increased skilled birth attendance, but failed to impact on either neonatal mortality or the caesarean section rate. In places with no women's groups, the SDIP had no impact on utilisation outcomes or neonatal mortality. The lack of any impact on neonatal mortality suggests that greater increases in utilisation or better quality of care are needed to improve health outcomes. The SDIP changed health care seeking behaviour only in those areas with women's groups highlighting the importance of effective communication of the policy to the wider public.


The potential of current- and wind-driven transport for environmental management of the Baltic Sea.
The ever increasing impact of the marine industry and transport on vulnerable sea areas puts the marine environment under exceptional pressure and calls for inspired methods for mitigating the impact of the related risks. We describe a method for preventive reduction of remote environmental risks caused by the shipping and maritime industry that are transported by surface currents and wind impact to the coasts. This method is based on characterizing systematically the damaging potential of the offshore areas in terms of potential transport to vulnerable regions of an oil spill or other pollution that has occurred in a particular area. The resulting maps of probabilities of pollution to be transported to the nearshore and the time it takes for the pollution to reach the nearshore are used to design environmentally optimized fairways for the Gulf of Finland, Baltic Proper, and south-western Baltic Sea.


fringe and Notch specify polar cell fate during Drosophila oogenesis.
fringe encodes a glycosyltransferase that modulates the ability of the Notch receptor to be activated by its ligands. We describe studies of fringe function during early stages of Drosophila oogenesis. Animals mutant for hypomorphic alleles of fringe contain follicles with an incorrect number of germline cells, which are separated by abnormally long and disorganized stalks. Analysis of clones of somatic cells mutant for a null allele of fringe localizes the requirement for fringe in follicle formation to the polar cells, and demonstrates that fringe is required for polar cell fate. Clones of cells mutant for Notch also lack polar cells and the requirement for Notch in follicle formation appears to map to the polar cells. Ectopic expression of fringe or of an activated form of Notch can generate an extra polar cell. Our results indicate that fringe plays a key role in positioning Notch activation during early oogenesis, and establish a function for the polar cells in separating germline cysts into individual follicles.


Cholesterol metabolism and therapeutic targets: rationale for targeting multiple metabolic pathways.
The liver is the major regulator of the plasma low density lipoprotein cholesterol (LDL-C) concentration because it is not only the site of formation of very low density lipoproteins (VLDL), the precursors of most LDL in the circulation, but it is also the organ where the bulk of receptor-mediated clearance of LDL takes place. The liver also initially clears all the cholesterol that is absorbed from the small intestine. The absorption of excess cholesterol can potentially increase the amount of cholesterol stored in the liver. This, in turn, can result in increased VLDL secretion, and hence LDL formation, and also downregulation of hepatic LDL receptor activity. Such events will potentially increase plasma LDL-C levels. The converse situation occurs when cholesterol absorption is inhibited. Cholesterol enters the lumen of the small intestine principally from bile and diet. The major steps involved in the absorption process have been characterized. On average, about half of all cholesterol entering the intestine is absorbed, but the fractional absorption rate varies greatly among individuals. While the basis for this variability is not understood, it may partly explain why some patients respond poorly or not at all to statins and other classes of lipid-lowering drugs. There are few data relating to racial differences in cholesterol absorption. One study reported a significantly higher rate in African Americans compared with non-African Americans. Multiple lipid-lowering drugs that target pathways involving the absorption, synthesis, transport, storage, catabolism, and excretion of cholesterol are available. Ezetimibe selectively blocks cholesterol absorption and lowers plasma LDL-C levels by an average of 18%. When ezetimibe is coadministered with lower doses of statins, there is an additive reduction in LDL-C level, which equals the reduction achieved with maximal doses of statins alone. Dual inhibition of cholesterol synthesis and absorption is an effective new strategy for treating hypercholesterolemia.


Post-exercise cutaneous hyperaemia resulting from local exercise of an extremity.
Large changes in skin blood flow occur after exercise. Most studies have concentrated on the systemic effects of vigorous exercise on skin blood flow. We were interested in the post-exercise response in the neighbourhood of focal exercise. We used a painless neuromuscular electronic stimulator to exercise the muscles of the forearm, producing flexion of the fingers. There was no change in blood pressure and only a small increase in heart rate during this exercise. We measured blood flow during a 5-min pre-exercise period and a 5-min post-exercise period at the forearm, at the dorsum of the index finger and on the pad of the index finger. We also measured values on the contralateral non-exercised extremity during exercise as well as during matched time periods in control experiments with no exercise. Exercise did elicit an increased blood flow in the post-exercise period at all three sites compared with the control experiments with no exercise and on the contralateral extremity. For example, the increase in blood flow at the finger dorsum was 2.1 +/- 0.1 ml (min 100 g)-1 after exercise compared with -0.08 +/- 0.09 ml min-1 100 g-1 during the control experiment and 0.1 +/- 0.1 ml (min 100 g)-1 on the contralateral arm (all P < 0.01). The local application of heat at the site of blood flow monitoring produced a substantial increase in the post-exercise response at the two finger locations [27.4 +/- 0.4 ml (min 100 g)-1 at the finger dorsum], but not at the arm. This is the first demonstration that highly focal exercise, unaccompanied by a systemic haemodynamic response, can elicit a post-exercise cutaneous hyperaemia. Local heating produced a large synergistic increase in the post-exercise hyperaemia at sites with arteriovenous microvascular perfusion but not at sites with primarily nutritive perfusion. These findings show that local vasoregulatory changes occur in response to exercise, even in the absence of whole-body haemodynamic and thermal change.


Pan-atlantic analysis of the overlap of a highly migratory species, the leatherback turtle, with pelagic longline fisheries.
Large oceanic migrants play important roles in ecosystems, yet many species are of conservation concern as a result of anthropogenic threats, of which incidental capture by fisheries is frequently identified. The last large populations of the leatherback turtle, Dermochelys coriacea, occur in the Atlantic Ocean, but interactions with industrial fisheries could jeopardize recent positive population trends, making bycatch mitigation a priority. Here, we perform the first pan-Atlantic analysis of spatio-temporal distribution of the leatherback turtle and ascertain overlap with longline fishing effort. Data suggest that the Atlantic probably consists of two regional management units: northern and southern (the latter including turtles breeding in South Africa). Although turtles and fisheries show highly diverse distributions, we highlight nine areas of high susceptibility to potential bycatch (four in the northern Atlantic and five in the southern/equatorial Atlantic) that are worthy of further targeted investigation and mitigation. These are reinforced by reports of leatherback bycatch at eight of these sites. International collaborative efforts are needed, especially from nations hosting regions where susceptibility to bycatch is likely to be high within their exclusive economic zone (northern Atlantic: Cape Verde, Gambia, Guinea Bissau, Mauritania, Senegal, Spain, USA and Western Sahara; southern Atlantic: Angola, Brazil, Namibia and UK) and from nations fishing in these high-susceptibility areas, including those located in international waters.


Effectiveness of different activated irrigation techniques on debris and smear layer removal from curved root canals: a SEM evaluation.
This study evaluates the effectiveness of different activated irrigation techniques on removal of debris and smear layer from curved root canals. Ninety mandibular molars with a root canal curvature between 20 and 40 degrees were assigned to 4 groups (n = 20): syringe irrigation (SI), passive ultrasonic activation (PUI), sonic activation with EDDY (ED) or EndoActivator (EA) and a control group. Mesiobuccal root canals were prepared to size 40, 0.04 and irrigated with NaOCl (3%) according to the respective technique. Roots were split longitudinally and subjected to scanning electron microscopic analysis. Presence of debris and smear layer was evaluated using 5-grade scoring systems with 200× and 1000× magnification, respectively. Data were analysed with nonparametric analysis for ordinal longitudinal data (α = 5%). Activation of the irrigant significantly improved smear layer removal (P < 0.05). Regarding debris, only activation with EA and ED was significantly more effective than SI (P < 0.05). No activation technique was able to eliminate debris and smear layer completely from curved root canals.


Osteopontin drives KRAS-mutant lung adenocarcinoma.
Increased expression of osteopontin (secreted phosphoprotein 1, SPP1) is associated with aggressive human lung adenocarcinoma (LADC), but its function remains unknown. Our aim was to determine the role of SPP1 in smoking-induced LADC. We combined mouse models of tobacco carcinogen-induced LADC, of deficiency of endogenous Spp1 alleles, and of adoptive pulmonary macrophage reconstitution to map the expression of SPP1 and its receptors and determine its impact during carcinogenesis. Co-expression of Spp1 and mutant KrasG12C in benign cells was employed to investigate SPP1/KRAS interactions in oncogenesis. Finally, intratracheal adenovirus encoding Cre recombinase was delivered to LSL.KRASG12D mice lacking endogenous or overexpressing transgenic Spp1 alleles. SPP1 was overexpressed in experimental and human LADC and portended poor survival. In response to two different smoke carcinogens, Spp1-deficient mice developed fewer and smaller LADC with decreased cellular survival and angiogenesis. Both lung epithelial- and macrophage-secreted SPP1 drove tumor-associated inflammation, while epithelial SPP1 promoted early tumorigenesis by fostering the survival of KRAS-mutated cells. Finally, loss and overexpression of Spp1 was, respectively, protective and deleterious for mice harboring KRASG12D-driven LADC. Our data support that SPP1 is functionally involved in early stages of airway epithelial carcinogenesis driven by smoking and mutant KRAS and may present an important therapeutic target.


Effect of automatic record keeping on vigilance and record keeping time.
We have evaluated the effect of an automatic anaesthesia record keeper (AARK) on record keeping time and vigilance. With informed patient consent and institutional approval, we videotaped the attending anaesthetist and his/her immediate surroundings during 66 surgical procedures. Thirty-seven cases were charted manually and the remaining 29 were charted with a commercially available AARK. In order to evaluate vigilance, a physician examiner entered the operating room unannounced once during 33 of the manually charted cases and during 22 of the automatically charted cases and asked the anaesthetist to turn away from the monitors and recall the current value of eight patient physiological variables. The examiner recorded the recalled values and also the actual current monitor values of these variables. The videotapes were reviewed and the anaesthetist's intraoperative time was categorized into 15 predefined activities, including intraoperative anaesthesia record keeping time. We compared recalled and actual variable values to determine if the recalled values were within clinically relevant error limits. There was no statistical difference between the mean percentage case time spent recording manually (14.11 (SD 3.98)%) and automatically (12.39 (3.92)%). Moreover, use of the AARK did not significantly affect vigilance. Despite major advances in monitoring technology over the past 14 years, record keeping still occupies 10-15% of the anaesthetist's intraoperative time. It appears that in using an AARK, the anaesthetist reallocates intraoperative record keeping time from manual charting to dealing with problems in the anaesthetist machine interface caused by inadequate design.


A case-study of the experiences of junior medical officers in the emergency departments of a metropolitan hospital and rural hospital.
Increased exposure to post-graduate rural medical training is associated with increased likelihood of future rural practice. Training rotations in rural emergency departments provide a possible avenue for such exposure, but have been under-investigated. This study aimed to compare junior medical officers' emergency department experiences in a metropolitan and a rural hospital to inform rural health workforce initiatives. Mixed-method case-study design. Two 10-week periods in the respective emergency departments. Four junior medical officers at the rural site and 22 junior medical officers at the metropolitan hospital. Caseloads extracted from electronic medical records and training experience. Data were collected over 142 days. The average number of patients seen per day, per junior medical officer, was significantly higher at the rural hospital emergency department (7.2 patients per day) in comparison with the metropolitan hospital (4.3 patients per day). Junior medical officers at the rural hospital saw relatively more lower acuity patients. The seven junior medical officers who were interviewed provided consistently positive responses regarding their training experiences in both locations. This was particularly evident in the rural hospital and was attributed to one-on-one supervision. Most junior medical officers agreed that their expectations for support and learning opportunities were met and/or exceeded. However, junior medical officers reported feeling more supported at the rural hospital due to direct contact and communication with senior medical officers. Placement in a smaller hospital emergency department did not disadvantage the junior medical officers' training in this case-study and provided a positive rural training experience. These findings support workforce policies which encourage rural hospital emergency department training.


A general organocatalyst for direct alpha-functionalization of aldehydes: stereoselective C-C, C-N, C-F, C-Br, and C-S bond-forming reactions. Scope and mechanistic insights.
The development of a general organocatalyst for the alpha-functionalization of aldehydes, via an enamine intermediate, is presented. Based on optically active alpha,alpha-diarylprolinol silyl ethers, the scope and applications of this catalyst for the stereogenic formation of C-C, C-N, C-F, C-Br, and C-S bonds are outlined. The reactions all proceed in good to high yields and with excellent enantioselectivities. Furthermore, we will present mechanistic insight into the reaction course applying nonlinear effect studies, kinetic resolution, and computational investigations leading to an understanding of the properties of the alpha,alpha-diarylprolinol silyl ether catalysts.


Local interactions and the global organization of a two-phase flow in a branching tree.
The transport of liquid plugs in a microfluidic branching tree is studied experimentally. The global flow pattern can be either symmetric or asymmetric, with daughter plugs dividing in synchrony or asynchrony as a function of the driving flow rate and the network geometry. For trees with narrowing channels, the plugs always reach the exits even at low flow rates. In contrast, only one path is opened in networks with widening channels when the flow rate is low. This behavior is explained by a comparison of the pressure drop necessary to drive viscocapillary motion of plugs in straight channels with the nonlinear pressure variations as a plug passes a bifurcation. A model is built, which predicts that only narrowing networks can be fully filled, while widening networks can never be fully invaded by a two-phase flow.


Inpatient pulmonary embolism after elective primary total hip and knee arthroplasty in the United States.
The incidence of inpatient pulmonary embolism in patients who have elective primary hip and knee arthroplasty in the United States is unknown. Prior studies have included patients with cancer, trauma, or revisions. The goal of this study was to determine the incidence and risks of inpatient pulmonary embolism after elective arthroplasty by type of procedure. We used the 1998 to 2009 Healthcare Cost and Utilization Project Nationwide Inpatient Sample for this retrospective cohort study. Patients who were sixty years of age or older and underwent elective primary total hip or knee arthroplasty were included. The study variable was the type of arthroplasty: total hip, total knee, or two joints. Inpatient pulmonary embolism was the primary outcome; mortality was secondary. Logistic regression determined the adjusted odds ratios of inpatient pulmonary embolism by procedure, adjusting for age, sex, Charlson Comorbidity Index, atrial fibrillation, and surgical indication. Records represented 5,044,403 hospital discharges after primary total hip or knee arthroplasty. Total knee arthroplasty comprised 66% of the admissions. Less than 5% of patients had two joint procedures. The overall incidence of pulmonary embolism was 0.358% (95% confidence interval [CI], 0.338, 0.378). The incidence of pulmonary embolism differed by procedure and was highest among patients who had two-joint arthroplasty (0.777%; 95% CI, 0.677, 0.876), was lowest in recipients of total hip arthroplasty (0.201%; 95% CI, 0.179, 0.223), and was intermediate in patients who had total knee arthroplasty (0.400%; 95% CI, 0.377, 0.423). The adjusted odds ratios of pulmonary embolism in patients who had two joint procedures were 3.89 times higher than among patients who had total hip arthroplasty, controlling for other factors. Elective total knee arthroplasty is associated with a higher incidence and odds of inpatient pulmonary embolism than is total hip arthroplasty; multiple procedures pose the highest risk for pulmonary embolism and associated mortality.


Intraoperative mechanical ventilation practice in thoracic surgery patients and its association with postoperative pulmonary complications: results of a multicenter prospective observational study.
Intraoperative mechanical ventilation may influence postoperative pulmonary complications (PPCs). Current practice during thoracic surgery is not well described. This is a post-hoc analysis of the prospective multicenter cross-sectional LAS VEGAS study focusing on patients who underwent thoracic surgery. Consecutive adult patients receiving invasive ventilation during general anesthesia were included in a one-week period in 2013. Baseline characteristics, intraoperative and postoperative data were registered. PPCs were collected as composite endpoint until the 5th postoperative day. Patients were stratified into groups based on the use of one lung ventilation (OLV) or two lung ventilation (TLV), endoscopic vs. non-endoscopic approach and ARISCAT score risk for PPCs. Differences between subgroups were compared using χ2 or Fisher exact tests or Student's t-test. Kaplan-Meier estimates of the cumulative probability of development of PPC and hospital discharge were performed. Cox-proportional hazard models without adjustment for covariates were used to assess the effect of the subgroups on outcome. From 10,520 patients enrolled in the LAS VEGAS study, 302 patients underwent thoracic procedures and were analyzed. There were no differences in patient characteristics between OLV vs. TLV, or endoscopic vs. open surgery. Patients received VT of 7.4 ± 1.6 mL/kg, a PEEP of 3.5 ± 2.4 cmH2O, and driving pressure of 14.4 ± 4.6 cmH2O. Compared with TLV, patients receiving OLV had lower VT and higher peak, plateau and driving pressures, higher PEEP and respiratory rate, and received more recruitment maneuvers. There was no difference in the incidence of PPCs in OLV vs. TLV or in endoscopic vs. open procedures. Patients at high risk had a higher incidence of PPCs compared with patients at low risk (48.1% vs. 28.9%; hazard ratio, 1.95; 95% CI 1.05-3.61; p = 0.033). There was no difference in the incidence of severe PPCs. The in-hospital length of stay (LOS) was longer in patients who developed PPCs. Patients undergoing OLV, endoscopic procedures and at low risk for PPC had shorter LOS. PPCs occurred frequently and prolonged hospital LOS following thoracic surgery. Proportionally large tidal volumes and high driving pressure were commonly used in this sub-population. However, large RCTs are needed to confirm these findings. This trial was prospectively registered at the Clinical Trial Register (www.clinicaltrials.gov; NCT01601223 ; registered May 17, 2012.).


Expanding Self-Management Support From Chronic to Acute Care: Implications for Value.
In this article, I advocate the expansion of self-management support from chronic to acute care as a means of increasing the value of services and highlighting the value of occupational therapy. Like people with chronic conditions, clients with acute conditions (1) need to participate in their own care, (2) require support for participation, (3) engage in care outside of traditional medical behaviors, and (4) benefit from a focus of care that extends beyond discrete episodes. Self-management support can facilitate adherence, promote holistic conceptualizations of health, and address long-term outcomes and costs. Given that self-management support aligns with occupational therapy's philosophical roots, expansion of self-management support to acute care highlights the profession's contribution to health promotion in this practice area.


Quantitative PCR-based competitive index for high-throughput screening of Salmonella virulence factors.
Salmonella enterica serovar Typhimurium is an intracellular pathogen and a main cause of food-borne illness. In this study, a quantitative PCR (qPCR)-based competitive index (CI) method was developed to simultaneously compare the growth of multiple Salmonella strains. This method was applied to a mixture of 17 Salmonella mutants lacking regulator genes, and their survival ratios were compared based on expression of natural resistance-associated macrophage protein 1 (Nramp1). Nramp1, as a major host innate immune component, controls the intracellular replication of pathogens. Deletion strains containing unique DNA barcodes in place of regulator genes were mixed with the parental control, and the bacteria were inoculated into congenic mice differing only at Nramp1. Most of the deletion strains were outcompeted by wild-type bacteria in either mouse strain, and the lack of Nramp1 didn't increase the tested strain/parent control replication ratios. When the same collection of mutants was tested in congenic mouse-derived primary macrophages, a major Nramp1-expressing cell type, six strains (ΔhimD, ΔphoP/phoQ, ΔrpoE, ΔrpoS, ΔompR/envZ, and Δhfq strains) grew better in Nramp1(-/-) than in Nramp1(+/+) macrophages, suggesting that these six regulators may play roles in overcoming Nramp1-mediated bactericidal activity in primary macrophages. The discrepancy in survival of macrophages and that of mice suggests either that there are differences in macrophage populations or that other cell types expressing Nramp1 control Salmonella proliferation in the host. The method described allows competitive infection analysis to be carried out on complex mixtures of bacteria and provides high reproducibility from independent biological replicates.


T-cell growth factor gene: lack of expression in human T-cell leukemia-lymphoma virus-infected cells.
Activated mature T cells require T-cell growth factor (TCGF) for continuous proliferation. However, many mature T cells infected with human T-cell leukemia-lymphoma virus grow independently of exogenously added TCGF. It is now reported that cells infected with this virus also lack detectable TCGF messenger RNA (less than one copy per cell) and thus do not produce their own growth factor. The results apparently rule out an autostimulation mechanism of growth control.


Taking apart the two-dimensional infrared vibrational echo spectra: more information and elimination of distortions.
Ultrafast two-dimensional infrared (2D-IR) vibrational echo spectroscopy can probe the fast structural evolution of molecular systems under thermal equilibrium conditions. Structural dynamics are tracked by observing the time evolution of the 2D-IR spectrum, which is caused by frequency fluctuations of vibrational mode(s) excited during the experiment. However, there are a variety of effects that can produce line shape distortions and prevent the correct determination of the frequency-frequency correlation function (FFCF), which describes the frequency fluctuations and connects the experimental observables to a molecular level depiction of dynamics. In addition, it can be useful to analyze different parts of the 2D spectrum to determine if dynamics are different for subensembles of molecules that have different initial absorption frequencies in the inhomogeneously broadened absorption line. Here, an important extension to a theoretical method for extraction of the FFCF from 2D-IR spectra is described. The experimental observable is the center line slope (CLSomega(m)) of the 2D-IR spectrum. The CLSomega(m) is obtained by taking slices through the 2D spectrum parallel to the detection frequency axis (omega(m)). Each slice is a spectrum. The slope of the line connecting the frequencies of the maxima of the sliced spectra is the CLSomega(m). The change in slope of the CLSomega(m) as a function of time is directly related to the FFCF and can be used to obtain the complete FFCF. CLSomega(m) is immune to line shape distortions caused by destructive interference between bands arising from vibrational echo emission, from the 0-1 vibrational transition (positive), and from the 1-2 vibrational transition (negative) in the 2D-IR spectrum. The immunity to the destructive interference enables the CLSomega(m) method to compare different parts of the bands as well as comparing the 0-1 and 1-2 bands. Also, line shape distortions caused by solvent background absorption and finite pulse durations do not affect the determination of the FFCF with the CLSomega(m) method. The CLSomega(m) can also provide information on the cross correlation between frequency fluctuations of the 0-1 and 1-2 vibrational transitions.


Uncertainty budgeting in fold change determination and implications for non-targeted metabolomics studies in model systems.
The p-value is the most prominent established metric for statistical significance in non-targeted metabolomics. However, its adequacy has repeatedly been the subject of discussion criticizing its uncertainty and its dependence on sample size and statistical power. These issues compromise non-targeted metabolomics in model systems, where studies typically investigate 5-10 samples per group. In this paper we propose a different approach for assessing the relevance of fold change (FC) data, where the FC is treated as a quantitative value and is validated by uncertainty budgeting. For the purpose of large-scale application in non-targeted metabolomics, we present a simplified approach for uncertainty propagation using experimental standard deviations of metabolite intensities as type A-summarized standard uncertainties. The resulting expanded FC uncertainty can be used to derive a minimum relevant FC as a complementary criterion in metabolomics data evaluation. This concept overcomes the need for a uniform p-value cut-off for all metabolites by considering the experimental uncertainty for each metabolite individually. The proposed procedure is part of analytical method validation, however the concept has not previously been applied to non-targeted metabolomics. A case study on mesenchymal stem cells cultured in normoxia and hypoxia demonstrates the practical value of this approach, in particular for studies with a small sample size. An online two-dimensional LC method coupled to mass spectrometry was crucial in providing both broad metabolome coverage and excellent experimental precision (<8% CV for peak areas, on average 0.5% CV for retention times) that was required for sensitive differential analysis as low as FC 1.1.


The proteomic response of sea squirts (genus Ciona) to acute heat stress: a global perspective on the thermal stability of proteins.
Congeners belonging to the genus Ciona have disparate distributions limited by temperature. Ciona intestinalis is more widespread with a cosmopolitan distribution ranging from tropical to sub-arctic zones, while Ciona savignyi is limited to temperate-latitudes of the northern Pacific Ocean. To compare the heat stress response between congeners, we quantified changes in protein expression using proteomics. Animals were exposed to 22°C, 25°C, and 28°C for 6h, then recovered at a control temperature (13°C) for 16h (high heat stress experiment). In a second experiment we exposed animals to lower levels of heat stress at 18°C, 20°C, and 23°C, with a 16°C control. A quantitative analysis, using 2D gel electrophoresis and MALDI-TOF/TOF mass spectrometry (with a 69% and 93% identification rate for Ciona intestinalis and Ciona savignyi, respectively), showed changes in a number of protein functional groups, including molecular chaperones, extracellular matrix proteins, calcium-binding proteins, cytoskeletal proteins and proteins involved in energy metabolism. Our results indicate that C. intestinalis maintains higher constitutive levels of molecular chaperones than C. savignyi, suggesting that it is prepared to respond faster to thermal stress. Systematic discrepancies between estimated versus predicted molecular masses of identified proteins differed between protein families and were more pronounced under high heat conditions, suggesting that thermal sensitivities are lower for cytoskeletal proteins and ATP-synthase than for any other protein group represented on 2D gels.


Epidemiology and Effects of Bacterial Infections in Patients With Cirrhosis Worldwide.
Bacterial infections are common and life-threatening in patients with cirrhosis. Little is known about the epidemiology of bacterial infections in different regions. We performed a multicenter prospective intercontinental study to assess the prevalence and outcomes of bacterial and fungal infections in patients with cirrhosis. We collected data from 1302 hospitalized patients with cirrhosis and bacterial or fungal infections at 46 centers (15 in Asia, 15 in Europe, 11 in South America, and 5 in North America) from October 2015 through September 2016. We obtained demographic, clinical, microbiology, and treatment data at time of diagnosis of infection and during hospitalization. Patients were followed until death, liver transplantation, or discharge. The global prevalence of multidrug-resistant (MDR) bacteria was 34% (95% confidence interval 31%-37%). The prevalence of MDR bacteria differed significantly among geographic areas, with the greatest prevalence in Asia. Independent risk factors for infection with MDR bacteria were infection in Asia (particularly in India), use of antibiotics in the 3 months before hospitalization, prior health care exposure, and site of infection. Infections caused by MDR bacteria were associated with a lower rate of resolution of infection, a higher incidence of shock and new organ failures, and higher in-hospital mortality than those caused by non-MDR bacteria. Administration of adequate empirical antibiotic treatment was independently associated with improved in-hospital and 28-day survival. In a worldwide study of hospitalized patients, we found a high prevalence of infection with MDR bacteria in patients with cirrhosis. Differences in the prevalence of MDR bacterial infections in different global regions indicate the need for different empirical antibiotic strategies in different continents and countries. While we await new antibiotics, effort should be made to decrease the spread of MDR bacteria in patients with cirrhosis.


Minimal bystander activation of CD8 T cells during the virus-induced polyclonal T cell response.
Acute infections with viruses such as lymphocytic choriomeningitis virus (LCMV) are associated with a massive polyclonal T cell response, but the specificities of only a small percentage of these activated T cells are known. To determine if bystander stimulation of T cells not specific to the virus plays a role in this T cell response, we examined two different systems, HY-specific T cell receptor (TCR)-transgenic mice, which have a restricted TCR repertoire, and LCMV-carrier mice, which are tolerant to LCMV. LCMV infection of HY-transgenic C57BL/6 mice induced antiviral CTLs that lysed target cells coated with two of the three immunodominant epitopes previously defined for LCMV (glycoprotein 33 and nucleoprotein 397). Although LCMV-induced cytotoxic T lymphocytes (CTLs) from C57BL/6 mice could lyse uninfected H-2(k) and H-2(d) allogeneic targets, LCMV-induced CTLs from HY mice lysed only the H-2(k)-expressing cells. The HY mice generated both anti-H-2(k) and anti-H-2(d) CTL in mixed leukocyte reactions, providing evidence that the generation of allospecific CTLs during acute LCMV infection is antigen specific. During the LCMV infection there was blastogenesis of the CD8+ T cell population, but the HY-specific T cells (as determined by expression of the TCR-alpha chain) remained small in size. To examine the potential for bystander stimulation under conditions of a very strong CTL response, T cell chimeras were made between normal and HY mice. Even in the context of a normal virus-induced CTL response, no stimulation of HY-specific T cells was observed, and HY-specific cells were diluted in number by day 9 after infection. In LCMV-carrier mice in which donor and host T cells could be distinguished by Thy1 allotypic markers, adoptive transfer of LCMV-immune T cells into LCMV-carrier mice, whose T cells were tolerant to LCMV, resulted in activation and proliferation of donor CD8 cells, but little or no activation of host CD8 cells. These results support the hypothesis that the massive polyclonal CTL response to LCMV infection is virus-specific and that bystander activation of non-virus-specific T cells is not a significant component of this response.


The Value of Learning about Natural History in Biodiversity Markets.
Markets for biodiversity have generated much controversy because of the often unstated and untested assumptions included in transactions rules. Simple trading rules are favored to reduce transaction costs, but others have argued that this leads to markets that favor development and erode biodiversity. Here, I describe how embracing complexity and uncertainty within a tradable credit system for the Red-cockaded Woodpecker (Picoides borealis) creates opportunities to achieve financial and conservation goals simultaneously. Reversing the effects of habitat fragmentation is one of the main reasons for developing markets. I include uncertainty in habitat fragmentation effects by evaluating market transactions using five alternative dispersal models that were able to approximate observed patterns of occupancy and movement. Further, because dispersal habitat is often not included in market transactions, I contrast how changes in breeding versus dispersal habitat affect credit values. I use an individually-based, spatially-explicit population model for the Red-cockaded Woodpecker (Picoides borealis) to predict spatial- and temporal- influences of landscape change on species occurrence and genetic diversity. Results indicated that the probability of no net loss of abundance and genetic diversity responded differently to the transient dynamics in breeding and dispersal habitat. Trades that do not violate the abundance cap may simultaneously violate the cap for the erosion of genetic diversity. To highlight how economic incentives may help reduce uncertainty, I demonstrate tradeoffs between the value of tradable credits and the value of information needed to predict the influence of habitat trades on population viability. For the trade with the greatest uncertainty regarding the change in habitat fragmentation, I estimate that the value of using 13-years of data to reduce uncertainty in dispersal behaviors is $6.2 million. Future guidance for biodiversity markets should at least encourage the use of spatially- and temporally-explicit techniques that include population genetic estimates and the influence of uncertainty.


Child safety seat counseling: three keys to safety.
The number one cause of death for children younger than 14 years is vehicular injury. Child safety seats and automobile safety belts protect children in a crash if they are used correctly, but if a child does not fit in the restraint correctly, it can lead to injury. A child safety seat should be used until the child correctly fits into an adult seat belt. It is important for physicians caring for children to know what child safety seats are available and which types of seats are safest. Three memory keys will help guide appropriate child safety seat choice: (1) Backwards is Best; (2) 20-40-80; and (3) Boost Until Big Enough. "Backwards is Best" cues the physician that infants are safest in a head-on crash when they are facing backward. "20-40-80" reminds the physician that children may need to transition to a different seat when they reach 20, 40, or 80 lb. "Boost Until Big Enough" emphasizes that children need to use booster seats until they are big enough to fit properly into an adult safety belt.


Palisaded neutrophilic granulomatous dermatitis in rheumatoid arthritis.
Palisaded neutrophilic granulomatous dermatitis (PNGD) is an entity that has not been clearly defined either clinically or histopathologically. It is seen in patients with rheumatoid arthritis and other connective tissue diseases. In the past, many cases of PNGD have been described under several different names including palisaded neutrophilic and granulomatous dermatitis, linear subcutaneous bands, interstitial granulomatous dermatitis with cutaneous cords and arthritis, rheumatoid papules, and Churg-Strauss granuloma. We report 7 additional cases of PNGD. Clinically, 6 patients presented with erythematous to violaceous plaques, papules, and nodules on multiple body sites; one presented with subcutaneous linear bands on the shoulder. Five had rheumatoid arthritis; one had adult-onset Still's disease; and one showed clinical signs of rheumatoid arthritis, although serologically the rheumatoid factor was negative. On histologic examination, a spectrum of changes was observed ranging from urticaria-like infiltrates to leukocytoclastic vasculitis and granuloma annulare with neutrophils. We report these cases to expand the histologic spectrum of this entity and to further delineate the different forms of clinical presentation.


Etanercept, a widely used inhibitor of tumor necrosis factor-α (TNF-α), prevents retinal ganglion cell loss in a rat model of glaucoma.
Visual loss in glaucoma is associated with pathological changes in retinal ganglion cell (RGC) axons and a slow decline in the RGC population. Age and elevated intraocular pressure (IOP) are the main risk factors for glaucomatous loss of vision. Several studies have implicated the proinflammatory cytokine tumor necrosis factor-α (TNF-α) as a link between elevated IOP and RGC death, but the cellular source of TNF-α and its causative role in RGC death remain uncertain. Here, using a rat model of glaucoma, we investigated the source of elevated TNF-α and examined whether Etanercept, a TNF-α blocker that is in common clinical use for other indications, is protective against RGC death. Episcleral vein cauterization (EVC) caused intraocular pressure (IOP) to be elevated for at least 28 days. IOP elevation resulted in a dramatic increase in TNF-α levels within a few days, axonal degeneration, and a 38% loss of RGCs by 4 weeks. Immunostaining coupled with confocal microscopy showed that OHT induced robust induction of TNF-α in Iba-1-positive microglia around the optic nerve head (ONH). Despite persistent elevation of IOP, Etanercept reduced microglial activation, TNF-α levels, axon degeneration in the optic nerve, and the loss of RGCs. Ocular hypertension (OHT) triggers an inflammatory response characterized by the appearance of activated microglia around the ONH that express TNF-α. Blocking TNF-α activity with a clinically approved agent inhibits this microglial response and prevents axonal degeneration and loss of RGCs. These findings suggest a new treatment strategy for glaucoma using TNF-α antagonists or suppressors of inflammation.


The Care of Adults with Intellectual Disabilities: Informal (Family) Caregivers' Perspectives.
The care of adults with intellectual disabilities is marginalized and rarely studied in Poland. In recent years, this issue has gained particular importance, partly due to the increasing life expectancy of people with ID. This paper presents a study of the function of informal caregivers for adults with ID, comprising parents who provide regular, constant, physical and emotional support and assistance with everyday activities to their adult children. Due to cultural and institutional conditions, Polish society sets high expectations for families regarding the care of their dependent members. Social policy also mainly promotes informal care, with formal care only being supported to a very limited extent. The state delegates responsibility, including financial responsibility, to families. With the rapid aging of society, this situation poses great challenges. This study was conducted in the Łodź region of Poland; it used a qualitative approach, and a semi-structured interview was performed using the narrative elements technique. The main goal of the research was to understand the situation of caregivers to adults with an intellectual disability by identifying thematic categories in the respondents' statements. The analysis of the qualitative data content made it possible to capture and present the participants' personal perspectives on significant issues connected with their function in the context of providing care to an adult with an intellectual disability. A total of 12 interviews were conducted. The age of the respondents (caregivers) was 51-82 years old, and the individuals they were caring for were between 20 and 49 years old. Based on the materials collected, 13 thematic categories and subcategories were identified, along with illustrative examples. The main categories concerned everyday functioning, health, uncertainty, relationships with others, feelings, time, and the macro level. For each category, subcategories were distinguished and illustrated by the respondents' statements. The categories and subcategories were not completely distinct; sometimes they overlapped or complemented one another. For the majority of the respondents, the care of an adult with an intellectual disability had a negative effect on their well-being. As a consequence, they performed their caregiver's role at the expense of their own lifestyle. Noticeable themes included "addiction" to caregiving, psychophysical fatigue, and the needs and difficulties resulting from this being "ignored" by the commonly understood social environment (including state institutions). Thus, the care of dependent adults with ID should be viewed on a broad human spectrum, that is, in consideration of the unique situation of those who remain under permanent care provided by family members, those who live alone, and those whose loved ones try to combine caregiving with their own private lives. This is becoming all the more important, as the number of seniors with intellectual disabilities will continue to grow in the coming decades.


Implementation of an oxygen concentrator system in district hospital paediatric wards throughout Malawi.
Hypoxaemia in children with severe or very severe pneumonia is a reliable predictor of mortality, yet oxygen was not available in most paediatric wards in Malawi. The Child Lung Health Programme in Malawi made oxygen available by supplying oxygen concentrators and essential supplies to 22 district and 3 regional hospitals' paediatric wards. Five key steps were taken to introduce concentrators: (1) develop a curriculum and training materials; (2) train staff on use and maintenance; (3) retrain electromedical departments on maintenance and repair; (4) conduct training once concentrators arrived in the country; and (5) distribute concentrators once staff had been trained. The paediatric wards in 3 regional and 22 government district hospitals and 3 regional electromedical engineering departments in Malawi. Main changes were: (1) provision of a source of oxygen in every paediatric ward in all district hospitals; (2) training of electrical engineering and health personnel in the use, maintenance and repair of oxygen concentrators; and (3) setting-up of high-dependency rooms or areas for severely ill children where oxygen is administered. It is feasible to implement an oxygen system using concentrators throughout a low-income country. Oxygen delivery requires trained staff with necessary equipment and supplies. Regular maintenance and supervision are essential to ensure optimal utilization.


Mutation in CEP63 co-segregating with developmental dyslexia in a Swedish family.
Developmental dyslexia is the most common learning disorder in children. Problems in reading and writing are likely due to a complex interaction of genetic and environmental factors, resulting in reduced power of studies of the genetic factors underlying developmental dyslexia. Our approach in the current study was to perform exome sequencing of affected and unaffected individuals within an extended pedigree with a familial form of developmental dyslexia. We identified a two-base mutation, causing a p.R229L amino acid substitution in the centrosomal protein 63 kDa (CEP63), co-segregating with developmental dyslexia in this pedigree. This mutation is novel, and predicted to be highly damaging for the function of the protein. 3D modelling suggested a distinct conformational change caused by the mutation. CEP63 is localised to the centrosome in eukaryotic cells and is required for maintaining normal centriole duplication and control of cell cycle progression. We found that a common polymorphism in the CEP63 gene had a significant association with brain white matter volume. The brain regions were partly overlapping with the previously reported region influenced by polymorphisms in the dyslexia susceptibility genes DYX1C1 and KIAA0319. We hypothesise that CEP63 is particularly important for brain development and might control the proliferation and migration of cells when those two events need to be highly coordinated.


Women's preferences for the delivery of the National Health Service Breast Screening Programme: a cross-sectional survey.
To elicit women's preferences for delivery of the National Health Service (NHS) Breast Screening Programme. Interview survey. Private households in the UK. Five hundred and ninety-seven women aged 45-75 interviewed as part of the National Statistics Omnibus Survey, December 2008 and January 2009. Preferred setting (hospital or community) and preferred type of screening unit (mobile or permanent). Rated importance of distance, time, convenience and cost of travelling to a screening unit. Forty per cent of women did not mind whether the screening unit was based in a community or hospital setting, and 52% did not mind whether the unit was permanent or mobile. Among those who expressed a preference, 59% preferred a hospital to a community setting, and 62% preferred a permanent to a mobile unit. Many women (63%) said distance to a breast screening unit was an important factor, as was time to travel (58%). Among those with access to a car, 80% regarded the availability of parking as important, but only 40% regarded the cost of car travel as important. Among women with no access to a car, there was a similar pattern for ease of public transport (76%) and cost of public transport (48%). Many women were unconcerned about the type and setting of breast screening units. Among those who were concerned, most preferred hospital over community settings and permanent over mobile units, but nonetheless most women said time and distance were important. Well-situated units with advanced publicity about public transport links and parking facilities may encourage greater uptake.


Mylar and Teflon-AF as cell culture substrates for studying endothelial cell adhesion.
The textured and opaque nature of Dacron and ePTFE has prevented the use of these fabrics in conventional cell culture techniques normally employed to optimize cell attachment and retention. This lack of optimization has led, in part, to the poor performance of endothelialization strategies for improving vascular graft patency. Here we show that thin, transparent films of Mylar and Teflon-AF are viable in vitro cell culture mimics of Dacron and ePTFE vascular graft materials, particularly for the study of protein mediated endothelial cell (EC) attachment, spreading and adhesion. Glass substrates were used as controls. X-ray photoelectron spectroscopy (XPS) and contact angle analysis showed that Mylar and Teflon-AF have surface chemistries that closely match Dacron and ePTFE. (125)I radiolabeling was used to quantify fibronectin (FN) adsorption, and FN and biotinylated-BSA "dual ligand" co-adsorption onto glass, Mylar and Teflon-AF substrates. Native human umbilical vein endothelial cells (HUVEC) and streptavidin-incubated biotinylated-HUVEC (SA-b-HUVEC) spreading was measured using phase contrast microscopy. Cell retention and adhesion was determined using phase contrast microscopy under laminar flow. All surfaces lacking protein pre-treatment, regardless of surface type, showed the lowest degree of cell spreading and retention. Dual ligand treated Mylar films showed significantly greater SA-b-HUVEC spreading up to 2 h, but were similar to HUVEC on FN treated Mylar at longer times; whereas SA-b-HUVEC spreading on dual ligand treated Teflon-AF was never significantly different from HUVEC on FN treated Teflon-AF at any time point. SA-b-HUVEC retention was significantly greater on dual ligand treated Mylar compared to HUVEC on FN treated Mylar over the entire range of shear stresses tested (3.54-28.3 dynes/cm(2)); whereas SA-b-HUVEC retention to dual ligand and HUVEC retention to FN treated Teflon-AF gave similar results at each shear stress, with only the mid-range of stresses showing significant difference in cell retention to Teflon-AF.


Coastal Zone Color Scanner atmospheric correction algorithm: multiple scattering effects.
An analysis of the errors due to multiple scattering which are expected to be encountered in application of the current Coastal Zone Color Scanner (CZCS) atmospheric correction algorithm is presented in detail. This was prompted by the observations of others that significant errors would be encountered if the present algorithm were applied to a hypothetical instrument possessing higher radiometric sensitivity than the present CZCS. This study provides CZCS users sufficient information with which to judge the efficacy of the current algorithm with the current sensor and enables them to estimate the impact of the algorithm-induced errors on their applications in a variety of situations. The greatest source of error is the assumption that the molecular and aerosol contributions to the total radiance observed at the sensor can be computed separately. This leads to the requirement that a value epsilon'(lambda,lambda(0)) for the atmospheric correction parameter, which bears little resemblance to its theoretically meaningful counterpart, must usually be employed in the algorithm to obtain an accurate atmospheric correction. The behavior of '(lambda,lambda(0)) with the aerosol optical thickness and aerosol phase function is thoroughly investigated through realistic modeling of radiative transfer in a stratified atmosphere over a Fresnel reflecting ocean. A unique feature of the analysis is that it is carried out in scan coordinates rather than typical earth-sun coordinates allowing elucidation of the errors along typical CZCS scan lines; this is important since, in the normal application of the algorithm, it is assumed that the same value of can be used for an entire CZCS scene or at least for a reasonably large subscene. Two types of variation of ' are found in models for which it would be constant in the single scattering approximation: (1) variation with scan angle in scenes in which a relatively large portion of the aerosol scattering phase function would be examined by the sensor in the single scattering approximation and (2) variation with aerosol optical thickness in a manner that increases with increasing solar zenith angle. In the worst case examined, the error associated with the variation of ' with scan angle was found to be 2.7-5.4 counts in Band 1 (depending on the turbidity of the atmosphere) for a marine aerosol, while the error associated with the variation of ' with aerosol optical thickness was at most 3 counts but would be reduced to negligible values when epsilon' could be determined in regions of high aerosol optical thickness. Since the water-leaving radiance must be determined with an accuracy of approximately 1-2 digital counts for maximum usefulness, these worst-case errors indicate that typically the algorithm will perform with the required accuracy in the case of CZCS, if limited to subscenes which are not too large. However, since for a variety of reasons it is highly desirable to be able to estimate the value of ' at each pixel, computations were performed to determine how accurately the algorithm would perform in retrieving the water-leaving radiance in the blue, assuming that it was known in the green and red. It is found that the simple expediency of decreasing the derived value of e' in the blue by 5% was sufficent to decrease the error in the retrieved water-leaving radiance to >2 counts for a variety of aerosol phase functions and aerosol optical thicknesses (including mildly absorbing aerosols) and for several orbit geometries. Thus we conclude that in situations where ' can be estimated at each pixel, this modification will result in water-leaving radiances with the desired accuracy in most cases.


Nanoscale thermometry by scanning thermal microscopy.
Measuring temperature is a central challenge in nanoscience and technology. Addressing this challenge, we report the development of a high-vacuum scanning thermal microscope and a method for non-equilibrium scanning probe thermometry. The microscope is built inside an electromagnetically shielded, temperature-stabilized laboratory and features nanoscopic spatial resolution at sub-nanoWatt heat flux sensitivity. The method is a dual signal-sensing technique inferring temperature by probing a total steady-state heat flux simultaneously to a temporally modulated heat flux signal between a self-heated scanning probe sensor and a sample. Contact-related artifacts, which so far limit the reliability of nanoscopic temperature measurements by scanning thermal microscopy, are minimized. We characterize the microscope's performance and demonstrate the benefits of the new thermometry approach by studying hot spots near lithographically defined constrictions in a self-heated metal interconnect.


Maternal hepcidin determines embryo iron homeostasis in mice.
Iron disorders are associated with adverse pregnancy outcomes, yet iron homeostatic mechanisms during pregnancy are poorly understood. In humans and rodents, the iron-regulatory hormone hepcidin is profoundly decreased in pregnant mothers, which is thought to ensure adequate iron availability for transfer across placenta. However, the fetal liver also produces hepcidin, which may regulate fetal iron endowment by controlling placental iron export. To determine the relative contribution of maternal vs embryo hepcidin to the control of embryo iron endowment in iron-sufficient or iron-overloaded mice, we generated combinations of mothers and embryos that had or lacked hepcidin. We found that maternal, but not embryonic, hepcidin determined embryo and placental iron endowment in a healthy pregnancy. We further determined that inflammation can counteract pregnancy-dependent suppression of maternal hepcidin. To establish how essential maternal hepcidin suppression is for embryo iron homeostasis, we mimicked the range of maternal hepcidin activity by administering a hepcidin peptide mimetic to pregnant mice. This also allowed us to determine the effect of isolated maternal hepcidin excess on pregnancy, in the absence of other confounding effects of inflammation. Higher doses of hepcidin agonist caused maternal iron restriction and anemia, lower placenta and embryo weight, embryo anemia, and increased embryo mortality. Low agonist doses did not cause maternal anemia but still adversely affected the embryo, causing anemia, tissue iron deficiency (including in the brain), and decreased weight. Our studies demonstrate that suppression of maternal hepcidin during pregnancy is essential for maternal and embryo iron homeostasis and health.


Risk of Gastrointestinal Bleeding Among Dabigatran Users - A Self Controlled Case Series Analysis.
This article aims to evaluate the real world risk of gastrointestinal bleeding among users naïve to dabigatran. We adopted a self-controlled case series design. We sampled 1215 eligible adult participants who were continuous insured users between July 1, 2010 and March 31, 2012 with use of dabigatran and at least one gastrointestinal bleeding episode. We used a conditional Poisson regression to estimate incidence rate ratios. The population consisted of 64.69% of male and 60.25% patients equal to or greater than age 65 at start of observation. After adjustment for time-variant confounders, the incidence rate of gastrointestinal bleeding was similar during dabigatran risk period and non-exposed period (incidence rate ratio [IRR] = 1.01, 95% confidence interval [CI] 0.90, 1.15). There was no significant difference in GI incidence rate between periods of dabigatran and warfarin (IRR = 0.99, 95% CI 0.75-1.31). Among this database of young and healthy participants, dabigatran was not associated with increased incidence rate of GI bleeding compared with non-exposed period among naïve dabigatran users. We did not detect an increased risk of GI bleeding over dabigatran vs warfarin risk period. Along with other studies on safety and effectiveness, this study should help clinicians choose the appropriate anticoagulant for their patients.


Statistical considerations and endpoints for clinical lung cancer studies: Can progression free survival (PFS) substitute overall survival (OS) as a valid endpoint in clinical trials for advanced non-small-cell lung cancer?
In the last decades significant progress has been achieved in the biological understanding of non-small-cell lung cancer (NSCLC) and its tumor heterogeneity has become more evident. The identification of novel tumor targets with different pathways has stimulated the search for anti-tumor agents with a specific target directed mode of action, stipulating the need of testing these agents in clinical trials with an appropriate choice of the study endpoint. Gold standard as an endpoint has been so far overall survival (OS). By definition there are 3 categories of classical endpoints applied generally in clinical lung cancer studies: survival time endpoints, symptom endpoints, and endpoints relying on patients' reporting. Beside classical endpoints like OS which are tending to show the direct clinical effect of treatment, efforts have been taken to substitute these classical endpoints by surrogates. As a surrogate candidate for OS progression-free survival (PFS) should have the inherent considerable advantage, that it can detect subpopulations with longer PFS intervals early. Based on the (sub-) population treated and having in mind the risk-benefit profile of the drug under consideration, PFS can be considered for regulatory decision making. If accompanied by some independent measures like quality of life or treatment toxicity, PFS should be able to cover the clinical benefit achieved by treatment. Selecting PFS as primary endpoint in Phase III trials of advanced NSCLC may be based on a number of questions such as: Does the definition of PFS fit into the setting used by other trials? Are there accepted consensus standards? Are there consistent surveillance intervals? Is validation for each agent group planned? Is the incremental improvement of PFS big enough (≥30%)? And are there some additional measures to confine clinical benefit? OS is still accepted as the gold standard in trials investigating advanced NSCLC. OS is easy to measure and precise but it may be difficult to interpret if treatment action takes place only in a small subinterval of overall survival. PFS with some additional measures has become attractive when it seems advisable to make study results available earlier. Candidates for supporting PFS as "additional measures" may be treatment toxicity and quality of life measures. PFS allows a more precise detection and attribution to effects of the investigational treatment without being compromised by subsequent treatments. Therefore "enriched PFS" can be considered as an alternative primary endpoint replacing OS in studies investigating advanced NSCLC. The endpoint selection process should always be performed carefully considering all true and surrogate endpoint options in respect to the hypotheses to be proven.


Corrosion behaviour of cobalt-chromium dental alloys doped with precious metals.
Precious metal based dental alloys generally exhibit a superior corrosion resistance, in particular enhanced resistance to pitting and crevice corrosion, compared to non-precious metal based alloys such as CoCr alloys. A new generation of Co-Cr alloys enriched with precious metals (Au, Pt, Ru) have now appeared on the market. The goal of this study was to clarify the effect of the precious metals additions on the corrosion behaviour of such alloys. Various commercial alloys with different doping levels were tested by electrochemical techniques in two different milieus based on the Fusayama artificial saliva and an electrolyte containing NaCl. Open circuit potentials, corrosion currents, polarization resistances, and crevices potentials were determined for the various alloys and completed by a coulometric analysis of the potentiodynamic curves. In addition, the microstructures were characterised by metallography and phase compositions analysed by EDX. The results show that the presence of precious metals can deteriorate the corrosion behaviour of Co-Cr alloys in a significant way. Gold doping, in particular, produces heterogeneous microstructures that are vulnerable to corrosive attack.


Impact of local vascular lesions assessed with optical coherence tomography and ablation points on blood pressure reduction after renal denervation.
Local vascular injury is detectable with optical coherence tomography (OCT) after catheter-based renal denervation (RDN). However, it is unclear whether the number and type of vascular lesions or the number of ablation points could affect blood pressure (BP) reduction. The aim of the study was to assess the impact of vascular injury induced by RDN detected with OCT and the number of ablation points on BP response after 1, 3 and 6 months. RDN was either performed with a Simplicity catheter or an EnligHTNTM multielectrode basket followed by OCT. BP was recorded prospectively as office measurement and 24-hour ambulatory blood pressure monitoring (24-h ABPM) at each time point. Correlations between type and number of vascular lesions, as well as ablation points, on BP reduction were performed. Out of 16 patients, two were lost to BP follow-up. We documented a BP reduction at 1, 3 and 6 months in both office and 24-h ABPM. The Δmean office systolic BP (SBP) reduction was -18.75 ± 24.55 mm Hg, -20.58 ± 16.92 mm Hg and -18.75 ± 29.39 mm Hg, respectively, and the Δmean 24h-ABPM SBP reduction was -6.50 ± 23.45 mm Hg, -16.88 ± 26.64 mm Hg and -13.89 ± 21.20 mm Hg, respectively. The number of vascular lesions did not correlate with office and 24h-SBP and diastolic BP reduction. However, there was a correlation between ablation points and office Δmean SBP reduction at 6 months (p <0.02). Our study demonstrates for the first time that the number and type of vascular lesions as assessed with OCT did not predict the success of BP reduction after RDN. However, we observed a substantial decrease in office SBP in relation to the number of ablation points at 6 months.


Exploring sexual risks in a forensic mental health hospital: perspectives from patients and nurses.
Patients utilising forensic mental health inpatient services experience a range of sexual risks, including vulnerability to sexual exploitation and exposure to sexually transmissible infections. However, there is a paucity of research exploring the issue of sexual risks from the standpoint of patients and the nurses who work closely with them in inpatient secure settings. This article presents findings from a qualitative exploratory study, which investigated the views of patients and nurses about sexual relationships in forensic mental health settings. Risk was a major theme arising from the data and is the focus of this article. Subthemes from nurse participants included sexual safety, sexual vulnerability, unplanned pregnancies, and male sexuality issues. Subthemes from patients included risks associated with sexual activity, access to information and sexual health care, unplanned pregnancies, vulnerability, and male sexuality issues. Knowledge about these sexual risks by patients and nurses were well articulated, however information and assistance were considered by patients to be less than satisfactory in improving their knowledge or in providing the support they considered important to reduce sexual risks. The issue of risk needs to be addressed, and nurses would be well placed to contribute; however they require education to improve their ability to provide sexual health education to patients along with strategies to ensure patients receive the support and services they require to reduce their exposure to sexual risks.


Doppler color flow imaging of carotid body tumors.
Doppler color flow imaging is a new ultrasound method for the simultaneous spatial display of tissue and vessel morphology (B-mode echotomograms) and of color-coded blood flow velocity information (Doppler-mode analysis). This new method is particularly useful in the neck, not only for the assessment of brain arteries but also for the safe and valid identification of carotid paragangliomas compared with other neck tumors. Early clinical detection of carotid paragangliomas is difficult since these lesions often occur sporadically and the patients remain symptom-free until the tumor becomes noticeable. Doppler color flow imaging allows the diagnosis of even small paragangliomas, which may improve management because of existing complications of surgical therapy.


A two-step optimization method for improving multiple brain lesion treatments with robotic radiosurgery.
Planning robotic radiosurgery treatments for multiple (n > 3) metastatic brain lesions is challenging due to the need of satisfying a large number of dose-volume constraints and the requirement of prescribing different dose levels to individual targets. In this study, we developed a sequential two-step optimization technique to improve the planning quality of such treatments. In contrast to the conventional approach of where all targets are simultaneously planned, we have developed a two-step optimization method. In this method, the first step was to create treatment plans for individual targets. In the second step, the 3D dose matrices associated with each plan were exported to Dicom-RT digital files and subsequently optimized. For the optimization, a singular-value-decomposition (SVD) algorithm was implemented to minimize the dose interferences among different targets. Finally, we compared the optimized treatment plans with the treatment plans created using the conventional method to determine the effectiveness of the new method. Large improvements in target dose distributions as well as normal brain sparing were found for the two-step optimization treatment plans as compared with the conventional treatment plans. The two-step optimization significantly lowered the volume of normal brain receiving relatively low doses. For example, the normal brain volume receiving 12-Gy was reduced by averaged 42% (range 34%-47%) with the two-step optimization. Such improvements generally enlarged with increasing number of targets being treated regardless of target sizes. Of note, normal brain dose was found to increase non-linearly with increasing number of targets. In summary, a two-step optimization technique is demonstrated to significantly improve the treatment plan quality as well as reduce the planning effort for multi-target robotic radiosurgery.


Morphology and origins of sedimentary structures on submarine slopes.
Submarine slopes in deep water, such as continental slopes, are often indented by valleys or channels and made uneven by ridges or levees. The origins of many of these features are unknown or disputed. Morphologically, however, there is often great similarity between forms on deep slopes and forms on shallow slopes or on land. Structurally the slopes in deep water are less well explored, but several observations reveal features, such as lamination and crossbedding, that are known from shallow water also. Measurements of current indicate that periodically the movement of water near the bottom is fast enough to move particles of sediment from time to time. Morphology, fine structure, and currents suggest that internal waves and associated currents, as well as gravity, may control the shape of deep submarine slopes analogously to the shaping by surface waves of slopes in shallow water.


Electrospun polyurethane as an alternative ventricular catheter and in vitro model of shunt obstruction.
Intracranial pressure and volume vary considerably between hydrocephalic patients, and with age, health and haemodynamic status; if left untreated, intracranial pressure rises and the ventricular system expands to accommodate the excess cerebrospinal fluid, with significant morbidity and mortality. Cerebrospinal fluid shunts in use today have a high incidence of failure with shunt obstruction being the most serious. Conventional proximal shunt catheters are made from poly(dimethyl)siloxane, the walls of which are perforated with holes for the cerebrospinal fluid to pass through. The limited range of catheters, in terms of material selection and flow distribution, is responsible in large part for their poor performance. In this study, we present an alternative design of proximal catheter made of electrospun polyether urethane, and evaluate its performance in the presence of glial cells, which are responsible for shunt blockage. The viability and growth of cells on catheter materials such as poly(dimethyl)siloxane and polyurethane in the form of cast films, microfibrous mats and porous sponges were studied in the presence of proteins present in cerebrospinal fluid after 48 h and 96 h in culture. The numbers of viable cells on each substrate were comparable to untreated poly(dimethyl)siloxane, both in the presence and absence of serum proteins found in cerebrospinal fluid. A cell culture model of shunt obstruction was developed in which cells on electrospun polyether urethane catheters were subjected to flow during culture in vitro, and the degree of obstruction quantified in terms of hydraulic permeability after static and perfusion culture. The results indicate that a catheter made of electrospun polyether urethane would be able to maintain cerebrospinal fluid flow even with the presence of cells for the time period chosen for this study. These findings have implications for the design and deployment of microporous shunt catheter systems for the treatment of hydrocephalus.


Medicine--a healing or a dying art?
That medicine is a science is the popular belief, and this has been reinforced by the advent of 'evidence-based medicine', which assumes that scientific proof is required to support medical practice. However, the view of science implied is a narrow one, foreign both to pure scientists and to artists, and the art of medicine is devalued by the approach. There are both important differences and important similarities between science and the arts. The arts should contribute to evidence-based practice and education along with science, and have a role in many aspects of medical practice.


Atypical presentation of sunflower epilepsy featuring an EEG pattern of continuous spike waves during slow-wave sleep.
Sunflower epilepsy is a rare photosensitive and commonly pharmacoresistant reflex epilepsy characterized by stereotyped seizures involving turning of the head towards light, similar to a sunflower turning towards the sun, and waving of the hands in front of the eyes, sometimes followed by absence seizures, myoclonic jerks, and tonic-clonic seizures. In the original description, seizures in sunflower epilepsy have been perceived as self-induced, but contemporary case series suggest that hand waving corresponds to an ictal phenomenon and not to a precipitating factor. We describe a nine-year-old girl featuring absence seizures with eye rolling or fluttering associated with hand waving movements. The chronological sequence of events based on a video-EEG-documented episode of our patient adds to the controversy surrounding the hypothesis of "self-induced" epileptic seizures in sunflower epilepsy. Shortly after epilepsy diagnosis, our patient presented with an EEG pattern of continuous spike waves in slow-wave sleep, an EEG feature that has not been described before and may relate to the cognitive deficit observed in some patients with sunflower epilepsy. Continuous spike waves in slow-wave sleep resolved, and lasting seizure freedom was achieved by a combination of ethosuximide and lamotrigine, which may be a possible alternative to valproic acid, particularly in girls and women of childbearing age. However, an attempt to taper anti-seizure drugs two years later led to seizure recurrence. We suggest performing sleep EEG recordings for sunflower epilepsy, particularly in patients with developmental stagnation or regression, to timely diagnose and treat continuous spike waves in slow-wave sleep syndrome.


Self-assembled peptide amphiphile nanofibers and peg composite hydrogels as tunable ECM mimetic microenvironment.
Natural extracellular matrix (ECM) consists of complex signals interacting with each other to organize cellular behavior and responses. This sophisticated microenvironment can be mimicked by advanced materials presenting essential biochemical and physical properties in a synergistic manner. In this work, we developed a facile fabrication method for a novel nanofibrous self-assembled peptide amphiphile (PA) and poly(ethylene glycol) (PEG) composite hydrogel system with independently tunable biochemical, mechanical, and physical cues without any chemical modification of polymer backbone or additional polymer processing techniques to create synthetic ECM analogues. This approach allows noninteracting modification of multiple niche properties (e.g., bioactive ligands, stiffness, porosity), since no covalent conjugation method was used to modify PEG monomers for incorporation of bioactivity and porosity. Combining the self-assembled PA nanofibers with a chemically cross-linked polymer network simply by facile mixing followed by photopolymerization resulted in the formation of porous bioactive hydrogel systems. The resulting porous network can be functionalized with desired bioactive signaling epitopes by simply altering the amino acid sequence of the self-assembling PA molecule. In addition, the mechanical properties of the composite system can be precisely controlled by changing the PEG concentration. Therefore, nanofibrous self-assembled PA/PEG composite hydrogels reported in this work can provide new opportunities as versatile synthetic mimics of ECM with independently tunable biological and mechanical properties for tissue engineering and regenerative medicine applications. In addition, such systems could provide useful tools for investigation of how complex niche cues influence cellular behavior and tissue formation both in two-dimensional and three-dimensional platforms.


Preconception Care in a Remote Aboriginal Community Context: What, When and by Whom?
Preconception care (PCC) is acknowledged as a vital preventive health measure aiming to promote health today and for subsequent generations. We aimed to describe the content and context of PCC delivery in a very remote Australian Aboriginal Community Controlled Health Service setting. A retrospective audit was undertaken to identify what PCC was delivered between 2011 and 2018 to 127 Aboriginal women who had at least one pregnancy during this period. Of 177 confirmed pregnancies, 121 had received PCC prior to the pregnancy. Sexually transmissible infection screening (71%) was the most common care delivered, followed by folic acid prescription (57%) and smoking cessation support (43%). Younger women received PCC less often, particularly screening for modifiable pregnancy risk factors. Rates of prediabetes/diabetes, albuminuria, overweight/obesity and smoking were high amongst those screened (48-60%). PCC was usually patient-initiated and increased significantly over the audit period. Presentation for antenatal care in the first trimester of pregnancy was high at 73%. Opportunities to increase PCC delivery include integration with routine health checks, pregnancy tests and chronic disease programs. PCC programs codesigned with young people are also recommended. All primary care providers should be supported and assisted to provide opportunistic PCC and health promotion.


Melatonin Suppresses the Growth of Ovarian Cancer Cell Lines (OVCAR-429 and PA-1) and Potentiates the Effect of G1 Arrest by Targeting CDKs.
Melatonin is found in animals as well as plants. In animals, it is a hormone that anticipates the daily onset of darkness and regulates physiological functions, such as sleep timing, blood pressure, and reproduction. Melatonin has also been found to have anti-tumor properties. Malignant cancers are the most common cause of death, and the mortality rate of ovarian tumor is the highest among gynecological diseases. This study investigated the anti-tumor effects of melatonin on the ovarian cancer lines, OVCAR-429 and PA-1. We observed the accumulation of melatonin-treated cells in the G₁ phase due to the down-regulation of CDK 2 and 4. Our results suggest that in addition to the known effects on prevention, melatonin may also provide anti-tumor activity in established ovarian cancer.


Exercise in patients with multiple sclerosis.
Exercise can be a beneficial rehabilitation strategy for people with multiple sclerosis to manage symptoms, restore function, optimise quality of life, promote wellness, and boost participation in activities of daily living. However, this population typically engages in low levels of health-promoting physical activity compared with adults from the general population, a fact which has not changed in the past 25 years despite growing evidence of the benefits of exercise. To overcome this challenge, the main limitations to promoting exercise through the patient-clinician interaction must be addressed. These limitations are the inadequate quality and scope of existing evidence, incomplete understanding of the mechanisms underlying the beneficial effects of exercise in people with multiple sclerosis, and the absence of a conceptual framework and toolkit for translating the evidence into practice. Future research to address those limitations will be essential to inform decisions about the inclusion of exercise in the clinical care of people with multiple sclerosis.


Use of an iron superoxide dismutase excreted by Trypanosoma cruzi in the diagnosis of Chagas disease: seroprevalence in rural zones of the state of Queretaro, Mexico.
Four iron superoxide dismutase (SODI, SODII, SODIII, and SODIV) activities with pI values of 6.9, 6.8, 5.25, and 3.8, respectively, were isolated from epimastigote forms of the Maracay strain of Trypanosoma cruzi cultivated at 28 degrees C in Grace's medium supplemented with 10% heat-inactivated fetal bovine serum. The activity of SODe (pI 3.8), which coincides with that of SODIV, was detected in Grace's medium without serum in which T. cruzi epimastigotes were cultivated for 24 hours at 28 degrees C. SODe, which was excreted into the medium by the parasite, was immunogenic and antibodies to SODe were detected in serum to a dilution of 1:2,500 by Western blot. The role of SODe is related to the establishment of the parasite within the host, and its high immunogenicity and specificity make it a useful molecular marker in diagnosing infection with this parasite. To validate a Western blot result using SODe as a antigen fraction, 1,029 sera of individuals from 11 municipalities in the state of Queretaro, Mexico were analyzed. Sampling was done randomly and results were compared with those for the same sera with three conventional serologic methods: an enzyme-linked immunosorbent assay (ELISA), indirect hemagglutination (IHA), and an indirect immunofluorescence assay (IFA) to detect antibodies to T. cruzi SODe. Samples that were positive by these three techniques were also positive by the Western blot method. The seroprevalence values for SODe (8.16% by ELISA and Western blot) in Queretaro were considerably higher than those reported in regions of Mexico considered to be endemic for Chagas disease. These results support the use of SODe in the serodiagnosis of Chagas disease.


Serpina1 is a potent inhibitor of IL-8-induced hematopoietic stem cell mobilization.
Here, we report that cytokine-induced (granulocyte colony-stimulating factor and IL-8) hematopoietic stem cell (HSC) and hematopoietic progenitor cell (HPC) mobilization is completely inhibited after low-dose (0.5 Gy) total-body irradiation (TBI). Because neutrophil granular proteases are regulatory mediators in cytokine-induced HSC/HPC mobilization, we considered a possible role for protease inhibitors in the induction of HSC/HPC mobilization. Bone marrow (BM) extracellular extracts that were obtained from murine femurs after 0.5 Gy of TBI contained an inhibitor of elastase. Also, after low-dose TBI, both Serpina1 mRNA and protein concentrations were increased in BM extracts, compared with extracts that were obtained from controls. The inhibitory activity in BM extracts of irradiated mice was reversed by addition of an Ab directed against Serpina1. To further study a possible in vivo role of Serpina1 in HSC/HPC mobilization, we administered Serpina1 before IL-8 injection. This administration resulted in an almost complete inhibition of HSC/HPC mobilization, whereas heat-inactivated Serpina1 had no effect. These results indicate that low-dose TBI inhibits cytokine-induced HSC/HPC mobilization and induces Serpina1 in the BM. Because exogenous administration of Serpina1 inhibits mobilization, we propose that radiation-induced Serpina1 is responsible for the inhibition of HSC/HPC mobilization. Also, we hypothesize that cytokine-induced HSC/HPC mobilization is determined by a critical balance between serine proteases and serine protease inhibitors.


N-acetyl-L-aspartyl-L-glutamate peptidase-like 2 is overexpressed in cancer and promotes a pro-migratory and pro-metastatic phenotype.
N-acetyl-L-aspartyl-L-glutamate peptidase-like 2 (NAALADL2) is a member of the glutamate carboxypeptidase II family, best characterized by prostate-specific membrane antigen (PSMA/NAALAD1). Using immunohistochemistry (IHC), we have shown overexpression of NAALADL2 in colon and prostate tumours when compared with benign tissue. In prostate cancer, NAALADL2 expression was associated with stage and Grade, as well as circulating mRNA levels of the NAALADL2 gene. Overexpression of NAALADL2 was shown to predict poor survival following radical prostatectomy. In contrast to PSMA/NAALAD1, NAALADL2 was localized at the basal cell surface where it promotes adhesion to extracellular matrix proteins. Using stable knockdown and overexpression cell lines, we have demonstrated NAALADL2-dependent changes in cell migration, invasion and colony-forming potential. Expression arrays of the knockdown and overexpression cell lines have identified nine genes that co-expressed with NAALADL2, which included membrane proteins and genes known to be androgen regulated, including the prostate cancer biomarkers AGR2 and SPON2. Androgen regulation was confirmed in a number of these genes, although NAALADL2 itself was not found to be androgen regulated. NAALADL2 was also found to regulate levels of Ser133 phosphorylated C-AMP-binding protein (CREB), a master regulator of a number of cellular processes involved in cancer development and progression. In combination, these data suggest that changes in expression of NAALADL2 can impact upon a number of pro-oncogenic pathways and processes, making it a useful biomarker for both diagnosis and prognosis.


Multiple Substrate Binding Mode-Guided Engineering of a Thermophilic PET Hydrolase.
Thermophilic polyester hydrolases (PES-H) have recently enabled biocatalytic recycling of the mass-produced synthetic polyester polyethylene terephthalate (PET), which has found widespread use in the packaging and textile industries. The growing demand for efficient PET hydrolases prompted us to solve high-resolution crystal structures of two metagenome-derived enzymes (PES-H1 and PES-H2) and notably also in complex with various PET substrate analogues. Structural analyses and computational modeling using molecular dynamics simulations provided an understanding of how product inhibition and multiple substrate binding modes influence key mechanistic steps of enzymatic PET hydrolysis. Key residues involved in substrate-binding and those identified previously as mutational hotspots in homologous enzymes were subjected to mutagenesis. At 72 °C, the L92F/Q94Y variant of PES-H1 exhibited 2.3-fold and 3.4-fold improved hydrolytic activity against amorphous PET films and pretreated real-world PET waste, respectively. The R204C/S250C variant of PES-H1 had a 6.4 °C higher melting temperature than the wild-type enzyme but retained similar hydrolytic activity. Under optimal reaction conditions, the L92F/Q94Y variant of PES-H1 hydrolyzed low-crystallinity PET materials 2.2-fold more efficiently than LCC ICCG, which was previously the most active PET hydrolase reported in the literature. This property makes the L92F/Q94Y variant of PES-H1 a good candidate for future applications in industrial plastic recycling processes.


Structure of a designed protein cage that self-assembles into a highly porous cube.
Natural proteins can be versatile building blocks for multimeric, self-assembling structures. Yet, creating protein-based assemblies with specific geometries and chemical properties remains challenging. Highly porous materials represent particularly interesting targets for designed assembly. Here, we utilize a strategy of fusing two natural protein oligomers using a continuous alpha-helical linker to design a novel protein that self assembles into a 750 kDa, 225 Å diameter, cube-shaped cage with large openings into a 130 Å diameter inner cavity. A crystal structure of the cage showed atomic-level agreement with the designed model, while electron microscopy, native mass spectrometry and small angle X-ray scattering revealed alternative assembly forms in solution. These studies show that accurate design of large porous assemblies with specific shapes is feasible, while further specificity improvements will probably require limiting flexibility to select against alternative forms. These results provide a foundation for the design of advanced materials with applications in bionanotechnology, nanomedicine and material sciences.


Rich Club Organization and Cognitive Performance in Healthy Older Participants.
The human brain is a complex network that has been noted to contain a group of densely interconnected hub regions. With a putative "rich club" of hubs hypothesized to play a central role in global integrative brain functioning, we assessed whether hub and rich club organizations are associated with cognitive performance in healthy participants and whether the rich club might be differentially involved in cognitive functions with a heavier dependence on global integration. A group of 30 relatively older participants (range = 39-79 years of age) underwent extensive neuropsychological testing, combined with diffusion-weighted magnetic resonance imaging to reconstruct individual structural brain networks. Rich club connectivity was found to be associated with general cognitive performance. More specifically, assessing the relationship between the rich club and performance in two specific cognitive domains, we found rich club connectivity to be differentially associated with attention/executive functions-known to rely on the integration of distributed brain areas-rather than with visuospatial/visuoperceptual functions, which have a more constrained neuroanatomical substrate. Our findings thus provide first empirical evidence of a relevant role played by the rich club in cognitive processes.


Assessment of dual-energy x-ray absorptiometry for use in evaluating the effects of dietary and environmental management on Hermann's tortoises (Testudo hermanni).
To assess dual-energy x-ray absorptiometry (DXA) for evaluating effects of diet and environment on bone mineral density in Hermann's tortoises (Testudo hermanni). 26 Hermann's tortoises within 1 month after hatching. Group 1 was housed in an artificial setting and fed naturally growing vegetation. Group 2 was housed in an artificial setting and fed vegetables grown for human consumption. Group 3 was maintained in an outside enclosure and fed naturally growing vegetation. After 10 months, pyramidal growth, body weight, and adverse conditions were assessed. Bone mineral density (BMD) of the axial and appendicular skeleton, shell, vertebral column, and pelvis was measured via DXA. Group 2 had the highest mean ± SD body weight (65.42 ± 30.85 g), followed by group 1 (51.08 ± 22.92 g) and group 3 (35.74 ± 7.13 g). Mean BMD of the shell varied significantly among groups (group 1, 0.05 ± 0.03 g/cm(2)•m; group 2, 0.09 ± 0.15 g/cm(2)•m; and group 3, undetectable). The BMD of the axial and appendicular skeleton, vertebral column, and pelvis did not differ significantly among groups. Pyramidal growth was highest in group 1 and not evident in group 3. Tortoises raised in artificial conditions did not have deficits in BMD, compared with results for outdoor-housed hibernating tortoises. Supplemental calcium was apparently not necessary when an adequate photothermal habitat and plant-based diet were provided. Higher BMD of captive-raised tortoises was morphologically associated with a higher incidence of pyramidal growth in captive-raised groups.


The Coventry Award. The effects of shelf life on clinical outcome for gamma sterilized polyethylene tibial components.
Tibial component shelf life was examined as a contributory factor of the in vivo failure of gamma sterilized prosthetic knee replacements. One hundred eighty-eight Synatomic total knee replacements sterilized by gamma irradiation in air were implanted by one surgeon into 147 patients between May 1985 and December 1994. Of these, 135 knees in 105 patients with a mean followup of 5.8 years (range, 2.1-11.3 years) were included in the study. The mean shelf life of the implants was 3.6 years (range, 0.1-10.7 years). Clinical failure for this study was defined as component retrieval resulting from polyethylene degradation. The knee components were divided into three different groups determined by their shelf storage durations of 0 to 4 years (Group 1, 93 components), between 4 and 8 years (Group 2, 21 components), or greater than 8 to 11 years (Group 3, 21 components). Six prostheses were revised because of polyethylene degradation after a mean implantation time of 2.5 years (range, 1.1-3.8 years). The mean shelf life of these six prostheses was 8.4 years (range, 5.8-9.6 years). Five years after implantation, prostheses that had shelf lives of less than 4 years had a 100% survival rate. Those that had shelf lives of 4 to 8 years before implantation had an 88.6% survival rate, and those prostheses that had shelf lives greater than 8 to 11 years had a 79.2% survival rate.


Role of Moringa oleifera on enterochromaffin cell count and serotonin content of experimental ulcer model.
The present study has been undertaken to observe the effect of aqueous extract of M. oleifera (MO) leaf (300mg/kg body weight) on mean ulcer index, enterochromaffin (EC) cells and serotonin (5-hydroxytryptamine; 5-HT) content of ulcerated gastric tissue. Ulceration was induced by using aspirin (500 mg/kg, po), cerebellar nodular lesion and applying cold stress. In all cases increased mean ulcer index in gastric tissue along with decreased EC cell count was observed with concomitant decrease of 5-HT content. Pretreatment with MO for 14 days decreased mean ulcer index, increased both EC cell count and 5-HT content in all ulcerated group, but treatment with ondansetron, a 5-HT3 receptor antagonist, along with MO pretreatment increased mean ulcer index, decreased 5-HT content without any alteration in EC cell count. The results suggest that the protective effect of MO on ulceration is mediated by increased EC cell count and 5-HT levels which may act via 5-HT3 receptors on gastric tissue.


A Statistical Test for Differential Item Pair Functioning.
This paper presents an IRT-based statistical test for differential item functioning (DIF). The test is developed for items conforming to the Rasch (Probabilistic models for some intelligence and attainment tests, The Danish Institute of Educational Research, Copenhagen, 1960) model but we will outline its extension to more complex IRT models. Its difference from the existing procedures is that DIF is defined in terms of the relative difficulties of pairs of items and not in terms of the difficulties of individual items. The argument is that the difficulty of an item is not identified from the observations, whereas the relative difficulties are. This leads to a test that is closely related to Lord's (Applications of item response theory to practical testing problems, Erlbaum, Hillsdale, 1980) test for item DIF albeit with a different and more correct interpretation. Illustrations with real and simulated data are provided.