Risk-adapted approach for fever and neutropenia in paediatric cancer patients--a national multicentre study.
In this national multicentre study, we examined the safety of reducing antibiotics in selected paediatric cancer patients with febrile neutropenia. Patients with signs of a bacterial infection and/or abnormal vital signs indicating sepsis were considered high risk and received antibiotic therapy. Remaining patients were allocated to low- or medium risk, depending on their interleukin-8 level. Low-risk patients did not receive any antibiotics and were discharged from the hospital after having been afebrile for 12 h. Medium-risk patients were re-evaluated after 72 h of antibiotic treatment and, in selected patients, antibiotics were stopped. Two hundred thirty-three febrile neutropenic episodes in 141 paediatric cancer patients were included in the study. Sixty-four episodes were classified high risk (28%), 122 medium risk (52%), and 47 (20%) low risk. In the medium-risk group, antibiotics were stopped after 72 h in 50 in 122 episodes (41%). Median duration of antibiotic treatment and hospital admission was significantly lower in low- and medium-risk episodes with early discharge. No failures were observed in the medium-risk group with early discharge. In the low-risk group, six failures were observed (12.8%), due to coagulase-negative staphylococci-positive blood cultures and recurrent fever. We showed that it is safe to shorten antibiotic treatment to 72 h in selected medium-risk patients with febrile neutropenia, regardless of the neutrophil count. The safety of withholding antibiotics in selected low-risk paediatric cancer patients with febrile neutropenia requires further investigation, using more suitable definitions for safety. Reduction in hospital admissions allows children with cancer more time at home and consequently improves their quality of life.


Measurement of a Cosmographic Distance Ratio with Galaxy and Cosmic Microwave Background Lensing.
We measure the gravitational lensing shear signal around dark matter halos hosting constant mass galaxies using light sources at z∼1 (background galaxies) and at the surface of last scattering at z∼1100 (the cosmic microwave background). The galaxy shear measurement uses data from the CFHTLenS survey, and the microwave background shear measurement uses data from the Planck satellite. The ratio of shears from these cross-correlations provides a purely geometric distance measurement across the longest possible cosmological lever arm. This is because the matter distribution around the halos, including uncertainties in galaxy bias and systematic errors such as miscentering, cancels in the ratio for halos in thin redshift slices. We measure this distance ratio in three different redshift slices of the constant mass (CMASS) sample and combine them to obtain a 17% measurement of the distance ratio, r=0.390_{-0.062}^{+0.070}, at an effective redshift of z=0.53. This is consistent with the predicted ratio from the Planck best-fit cold dark matter model with a cosmological constant cosmology of r=0.419.


Diminished humoral responses against and reduced gene expression levels of human endogenous retrovirus-K (HERV-K) in psoriasis.
Psoriasis is a multifactorial, chronic disease of skin affecting 2-3% of the world's population. Genetic studies of psoriasis have identified a number of susceptibility genes that are involved in anti-viral immunity. Furthermore, physiological studies have also found an increase in anti-viral proteins in psoriatic skin. These findings suggest the presence of an anti-viral state in psoriatic skin. However, the triggers for this anti-viral cascade and its consequences for host immunity are not known. Endogenous retroviruses have previously been described in many autoimmune diseases including psoriasis. In the present study we examined the humoral immune response against human endogenous retrovirus-K (HERV-K) proteins and the cutaneous expression levels of multiple HERV-K genes in psoriasis patients and healthy controls. In psoriatic sera we observed a significant decrease in IgM response against three HERV-K proteins: Env surface unit (SU), Env transmembrane protein (TM), and Gag capsid (CA) in comparison to sera obtained from blood bank healthy controls. A decrease in IgG response was also observed against CA. Furthermore, using quantitative RT-PCR we observed a decrease in the expression of HERV-K Env, Gag, Pol and Rec as well as ERV-9 genes in lesional psoriatic skin as compared to healthy skin. Together, our results suggest that the pro-inflammatory, anti-viral state in psoriasis is associated with diminished expression of HERV-K gene transcripts and a concomitant decrease in humoral responses to HERV-K. Our results indicate that a simple model where continuous, minimally changing HERV-K expression serves as an antigenic trigger in psoriasis might not be correct and further studies are needed to decipher the possible relationship between psoriasis and HERVs.


Implementing health research through academic and clinical partnerships: a realistic evaluation of the Collaborations for Leadership in Applied Health Research and Care (CLAHRC).
The English National Health Service has made a major investment in nine partnerships between higher education institutions and local health services called Collaborations for Leadership in Applied Health Research and Care (CLAHRC). They have been funded to increase capacity and capability to produce and implement research through sustained interactions between academics and health services. CLAHRCs provide a natural 'test bed' for exploring questions about research implementation within a partnership model of delivery. This protocol describes an externally funded evaluation that focuses on implementation mechanisms and processes within three CLAHRCs. It seeks to uncover what works, for whom, how, and in what circumstances. This study is a longitudinal three-phase, multi-method realistic evaluation, which deliberately aims to explore the boundaries around knowledge use in context. The evaluation funder wishes to see it conducted for the process of learning, not for judging performance. The study is underpinned by a conceptual framework that combines the Promoting Action on Research Implementation in Health Services and Knowledge to Action frameworks to reflect the complexities of implementation. Three participating CLARHCS will provide in-depth comparative case studies of research implementation using multiple data collection methods including interviews, observation, documents, and publicly available data to test and refine hypotheses over four rounds of data collection. We will test the wider applicability of emerging findings with a wider community using an interpretative forum. The idea that collaboration between academics and services might lead to more applicable health research that is actually used in practice is theoretically and intuitively appealing; however the evidence for it is limited. Our evaluation is designed to capture the processes and impacts of collaborative approaches for implementing research, and therefore should contribute to the evidence base about an increasingly popular (e.g., Mode two, integrated knowledge transfer, interactive research), but poorly understood approach to knowledge translation. Additionally we hope to develop approaches for evaluating implementation processes and impacts particularly with respect to integrated stakeholder involvement.


Update on the Pharmacological Treatment of Pathological Gambling.
This is an update to a previously published article discussing the neuropsychopharmacology of pathological gambling (PG) (1). In the prior manuscript, we described how cortico-limbic circuitry and neurotransmitter systems (norepinephrine, serotonin, dopamine, opioids, glutamate, and gamma-aminobutyric acid (GABA)) have been implicated in PG. These systems represent potential targets for psychopharmacological treatments for PG, with opioid antagonists arguably showing the most consistent benefit in RCTs. In the past year and half since this publication was prepared, there has been one additional randomized clinical trial (RCT) published along with a single case study. Our original manuscript did not describe in detail findings from case studies or open-label studies so in addition to the new RCT data and a new case report involving naltrexone, here we describe case and open-label findings. A PubMed search was conducted using terms such as "pathological gambling treatment", "clinical trials and gambling", and "gambling psychopharmacology." Using these search terms, numerous results were obtained, necessitating further search modifiers. For example, using just "pathological gambling treatment" results in over 1600 hits. In order to focus in on the search modalities, we searched within the initial results for specific phrases such as "psychopharmacology, clinical trial, medication, serotonergic, dopaminergic, etc." in addition to searching for specific medications. Results not directly related to the treatment of pathological gambling were not included. The study of pathological gambling is relatively new. As such, our search did not exclude any studies due to age of material, but with a few exceptions, the majority of the studies discussed were published later than 2000. This resulted in 24 case studies and/or RCTs not previously included in our original review article. These findings in conjunction with our prior publication provide a comprehensive overview of controlled investigations and exploratory reports of pharmacotherapies for PG.


Prospectively Isolated Tetraspanin+ Neoblasts Are Adult Pluripotent Stem Cells Underlying Planaria Regeneration.
Proliferating cells known as neoblasts include pluripotent stem cells (PSCs) that sustain tissue homeostasis and regeneration of lost body parts in planarians. However, the lack of markers to prospectively identify and isolate these adult PSCs has significantly hampered their characterization. We used single-cell RNA sequencing (scRNA-seq) and single-cell transplantation to address this long-standing issue. Large-scale scRNA-seq of sorted neoblasts unveiled a novel subtype of neoblast (Nb2) characterized by high levels of PIWI-1 mRNA and protein and marked by a conserved cell-surface protein-coding gene, tetraspanin 1 (tspan-1). tspan-1-positive cells survived sub-lethal irradiation, underwent clonal expansion to repopulate whole animals, and when purified with an anti-TSPAN-1 antibody, rescued the viability of lethally irradiated animals after single-cell transplantation. The first prospective isolation of an adult PSC bridges a conceptual dichotomy between functionally and molecularly defined neoblasts, shedding light on mechanisms governing in vivo pluripotency and a source of regeneration in animals. VIDEO ABSTRACT.


Free breakfasts in schools: design and conduct of a cluster randomised controlled trial of the Primary School Free Breakfast Initiative in Wales [ISRCTN18336527].
School-based breakfast provision is increasingly being seen as a means of improving educational performance and dietary behaviour amongst children. Furthermore, recognition is growing that breakfast provision offers potential as a means of addressing social inequalities in these outcomes. At present however, the evidence base on the effectiveness of breakfast provision in bringing about these improvements is limited. This paper describes the research design of a large scale evaluation of the effectiveness of the Welsh Assembly Government's Primary School Free Breakfast Initiative. A cluster randomised trial, with school as the unit of randomisation was used for the outcome evaluation, with a nested qualitative process evaluation. Quantitative outcome measures included dietary habits, attitudes, cognitive function, classroom behaviour, and school attendance. The study recruited 111 primary schools in Wales, of which 56 were randomly assigned to control condition and 55 to intervention. Participants were Year 5 and 6 students (aged 9-11 years) in these schools. Data were collected for all 111 schools at each of three time points: baseline, 4 month and 12 month follow-up. This was achieved through a repeated cross-sectional survey of approximately 4350 students on each of these occasions. Of those students in Year 5 at baseline, 1975 provided data at one or both of the follow-ups, forming a nested cohort. The evaluation also included a nested process evaluation, using questionnaires, semi-structured interviews and case studies with students, school staff, and local authority scheme coordinators as key informants. An overview of the methods used for the evaluation is presented, providing an example of the feasibility of conducting robust evaluations of policy initiatives using a randomised trial design with nested process evaluation. Details are provided of response rates and the flow of participants. Reflection is offered on methodological issues encountered at various stages through the course of the study, focusing upon issues associated with conducting a randomised trial of a government policy initiative, and with conducting research in school settings.


Probing the mechanisms underlying modulation of quinidine sensitivity to cardiac I(Ks) block by protein kinase A-mediated I(Ks) phosphorylation.
Cardiac I(Ks) is enhanced by protein kinase A (PKA) stimulation. And PKA-stimulated I(Ks) is about threefold less sensitive to quinidine block than basal current. In this study, we further tested two competing hypotheses: I(Ks) phosphorylation either (i) modulates access of blocking drugs to a binding site; or (ii) destabilizes the drug-channel interaction. To distinguish between these hypotheses, we studied quinidine block of I(Ks) channels in which three PKA site residues of the alpha-subunit KCNQ1 were mutated with a bulky negative charged aspartic acid (D). To study alleviation of I(Ks) block by quinidine, we compared activating current at +60 mV, either with or without 5 s hyperpolarizing prepulses to -120 mV. Without PKA stimulation, quinidine (100 microM) blocked wild-type current to a similar extent with and without the prepulse (93 +/- 2% of pre-drug current at +60 mV vs. 95 +/- 1%). With PKA-stimulated wild-type channels, however, there was less block with the hyperpolarization to -120 mV: at +60 mV, block was 71 +/- 2% (-prepulse) versus 58 +/- 3% (+prepulse). Individual D-mutations and the triple-D mutant were resistant to quinidine block similar to that seen with PKA-stimulated wild-type I(Ks). We conclude that phosphorylation-induced insertion of bulky negative charges alleviates quinidine block and that PKA-induced stimulation, by conferring negative charges to the channels, blunts I(Ks) block as the interaction between the channels and blockers becomes destabilized. These effects would be of clinical significance in providing protective mechanisms against pro-arrhythmias caused by drug-induced inhibition of I(Ks) and I(Kr).


Hypoxia Inducible Factor-2 Alpha and Prolinhydroxylase 2 Polymorphisms in Patients with Acute Respiratory Distress Syndrome (ARDS).
Hypoxia-inducible-factor-2α (HIF-2α) and HIF-2 degrading prolyl-hydroxylases (PHD) are key regulators of adaptive hypoxic responses i.e., in acute respiratory distress syndrome (ARDS). Specifically, functionally active genetic variants of HIF-2α (single nucleotide polymorphism (SNP) [ch2:46441523(hg18)]) and PHD2 (C/T; SNP rs516651 and T/C; SNP rs480902) are associated with improved adaptation to hypoxia i.e., in high-altitude residents. However, little is known about these SNPs' prevalence in Caucasians and impact on ARDS-outcome. Thus, we tested the hypotheses that in Caucasian ARDS patients SNPs in HIF-2α or PHD2 genes are (1) common, and (2) independent risk factors for 30-day mortality. After ethics-committee approval, 272 ARDS patients were prospectively included, genotyped for PHD2 (Taqman SNP Genotyping Assay) and HIF-2α-polymorphism (restriction digest + agarose-gel visualization), and genotype dependent 30-day mortality was analyzed using Kaplan-Meier-plots and multivariate Cox-regression analyses. Frequencies were 99.62% for homozygous HIF-2α CC-carriers (CG: 0.38%; GG: 0%), 2.3% for homozygous PHD2 SNP rs516651 TT-carriers (CT: 18.9%; CC: 78.8%), and 3.7% for homozygous PHD2 SNP rs480902 TT-carriers (CT: 43.9%; CC: 52.4%). PHD2 rs516651 TT-genotype in ARDS was independently associated with a 3.34 times greater mortality risk (OR 3.34, CI 1.09-10.22; p = 0.034) within 30-days, whereas the other SNPs had no significant impact (p = ns). The homozygous HIF-2α GG-genotype was not present in our Caucasian ARDS cohort; however PHD2 SNPs exist in Caucasians, and PHD2 rs516651 TT-genotype was associated with an increased 30-day mortality suggesting a relevance for adaptive responses in ARDS.


Densities of rectal peptide YY and somatostatin cells as biomarkers for the diagnosis of irritable bowel syndrome.
Irritable bowel syndrome (IBS) is a common chronic disorder. IBS diagnosis is a diagnosis of exclusion since there are no blood tests, radiological or endoscopic examinations for this disorder. Although several attempts have been made to develop a symptoms-based diagnosis, such systems are not widely used in clinics. Several tests and examinations measuring pathological findings in IBS have been considered for the diagnosis of IBS, but none of them has proved useful as a biomarker. Abnormalities in the cell densities of rectal peptide YY (PYY) and somatostatin cells have been reported in IBS patients. The aim of the present study was to determine the utility of these abnormalities as biomarkers for the diagnosis of IBS. Patients with IBS established according to Rome III criteria (n = 101) were included in this study (71 females and 30 males with a mean age of 35 years; range 18-61 years), and 62 healthy subjects (38 females and 24 males with a mean age of 41 years; range 18-65 years) were recruited as controls. Both the patients and controls underwent colonoscopy during which rectal biopsy samples were taken. The tissue samples were immunostained for PYY and somatostatin, and the number of stained cells was quantified relative to both the area of epithelial cells and per microscopic field. The density of PYY cells was significantly lower in IBS patients than in the healthy controls (P < 0.0001); receiver operator characteristic (ROC) analysis revealed an area under the ROC curve (AUC) of 0.99. The somatostatin cell density in IBS patients was higher than in the controls (P < 0.0001); ROC analysis revealed an AUC of 0.86. The densities of the rectal PYY and somatostatin cells appear to be clinically effective biomarkers for IBS. Furthermore, measurement of these parameters is inexpensive, rapid and does not require considerable experience or sophisticated equipment.


[Comparison of five video laryngoscopes and conventional direct laryngoscopy : Investigations on simple and simulated difficult airways on the intubation trainer].
Securing the airway with a tracheal tube is essential in emergency situations, in the intensive care setting as well as during anesthesia for surgery and other interventions. Current methods of airway assessment are poor screening tests for predicting difficult direct laryngoscopy due to a generally low positive predictive value; therefore, successful endotracheal intubation requires a high level of expertise, regular training and practice and sometimes additional tools. Currently, several video laryngoscopes (VL) with different designs are commercially available and have been investigated in a wide variety of settings. To our knowledge there is no prospective study evaluating and comparing performance among these three groups of VL; therefore, the aim of this study was to compare performance of five VL and conventional direct laryngoscopy in an intubation manikin model, both in a normal and simulated difficult intubation setting. In this study 10 residents, 12 senior staff physicians and 5 anesthesia nurses, all experienced in conventional direct laryngoscopy and inexperienced with VL underwent theoretical and hands-on training with all VL lasting 60 min. Afterwards participants performed intubation with all 5 VL and conventional direct laryngoscopy in a randomized sequence using an intubation manikin with normal intubation settings. Thereafter participants performed intubation in a simulated difficult intubation setting using the same intubation manikin with a neck collar to immobilize the cervical spine. In this study, the C-MAC(®) with Macintosh blade size 3, GlideScope(®) size 3, McGrath(®) series 5, King Vision(®) and Airtraq(®) size 2 were used. Time to intubation served as primary outcome and time to glottis visualization, number of intubation attempts, success rate and subjective evaluation of difficulty served as secondary outcomes. In the normal intubation setting, time to intubation ranged from 16.0 s (conventional direct laryngoscopy) to 34.3 s (McGrath). GlideScope and conventional direct laryngoscopy were successful in 100 % followed by C-MAC (96.7 %), Airtraq (88.9 %), King Vision (77.8 %) and McGrath VL (44.4 %). In the simulated difficult intubation setting, time to intubation ranged between 20.3 s (Airtraq) and 26.7 s (McGrath). Success rate with C-MAC was 100 %, followed by GlideScope (96.7 %), Airtraq (85.2 %), conventional direct laryngoscopy (85.2 %), King Vision (81.5 %) and McGrath VL (70.4 %). In the manikin with normal intubation setting, conventional direct laryngoscopy using a Macintosh blade was convincing and superior to any VL used in this study. During simulated difficult intubation, a blade with video transmission, such as C-MAC and the GlideScope were superior compared to conventional direct laryngoscopy and any other VL tested.


A geometric method for automatic extraction of sulcal fundi.
Sulcal fundi are 3-D curves that lie in the depths of the cerebral cortex and, in addition to their intrinsic value in brain research, are often used as landmarks for downstream computations in brain imaging. In this paper, we present a geometric algorithm that automatically extracts the sulcal fundi from magnetic resonance images and represents them as spline curves lying on the extracted triangular mesh representing the cortical surface. The input to our algorithm is a triangular mesh representation of an extracted cortical surface as computed by one of several available software packages for performing automated and semi-automated cortical surface extraction. Given this input we first compute a geometric depth measure for each triangle on the cortical surface mesh, and based on this information we extract sulcal regions by checking for connected regions exceeding a depth threshold. We then identify endpoints of each region and delineate the fundus by thinning the connected region while keeping the endpoints fixed. The curves, thus, defined are regularized using weighted splines on the surface mesh to yield high-quality representations of the sulcal fundi. We present the geometric framework and validate it with real data from human brains. Comparisons with expert-labeled sulcal fundi are part of this validation process.


Prognostic significance of elevated cardiac troponin-T levels in acute respiratory distress syndrome patients.
Elevated levels of biochemical markers of myocardial necrosis have been associated with worsened outcomes in Acute Respiratory Distress Syndrome (ARDS), but there are few prospective data on this relationship. We investigated elevated cardiac troponin T (cTnT) levels and their relationship with outcome in patients with ARDS. A prospective cohort study of patients with ARDS was conducted at a tertiary-care academic medical center. Patients had blood taken within 48 hours of ARDS onset and assayed for cTnT. Patients were followed for the outcomes of 60-day mortality, number of organ failures, and days free of mechanical ventilation. Echocardiographic and electrocardiographic (ECG) data were analyzed for signs of myocardial ischemia, infarction, or other myocardial dysfunction. 177 patients were enrolled, 70 of whom died (40%). 119 patients had detectable cTnT levels (67%). Median cTnT level was 0.03 ng/mL, IQR 0-0.10 ng/mL, and levels were higher among non-survivors (P = .008). Increasing cTnT level was significantly associated with increasing mortality (P = .008). The association between increasing cTnT level and mortality remained significant after adjustment in a multivariate model (HR(adj) = 1.45, 95% CI 1.17-1.81, P = .001). Elevated cTnT level was also associated with increased number of organ failures (P = .002), decreased number of days free of mechanical ventilation (P = .03), echocardiographic wall motion abnormalities (P = 0.001), and severity of tricuspid regurgitation (P = .04). There was no association between ECG findings of myocardial ischemia or infarction and elevated cTnT. Elevated cTnT levels are common in patients with ARDS, and are associated with worsened clinical outcomes and certain echocardiographic abnormalities. No association was seen between cTnT levels and ECG evidence of coronary ischemia.


The Effects of the Traditional Scaling Technique Versus a Modified Scaling Technique on Muscle Activity and Pinch Force Generation: A pilot study.
Purpose: Dental hygienists perform precision instrumentation tasks repetitively throughout the workday, placing them at increased risk for developing a musculoskeletal disorder. The purpose of this pilot study was to determine differences in muscle activity and pinch force generation between the traditional scaling technique and a modified scaling technique.Methods: A convenience sample of dental hygienists (n=12) acted as their own controls in this counterbalance-designed pilot study. Muscle activity and pinch forces were assessed while participants performed traditional and modified scaling techniques with designated instruments on artificial calculus applied to the lower left quadrant of a typodont, for a period of five minutes. Surface electromyography was used to measure muscle activity; sensors attached to the instrument handle measured pinch forces. Participants were surveyed regarding the instruments used and scaling technique preferences at the conclusion of the session. Parametric and non-parametric tests were used to analyze the data. Descriptive statistics were used to analyze the exit survey.Results: The modified scaling technique required less muscle activity than the traditional technique while scaling, however results were not significant (p>0.05). The traditional scaling technique required greater overall pinch force during scaling (p=.00). Pairwise comparisons revealed significant differences between pinch force generation in the thumb for the two scaling techniques (Z = -2.401, p= 0.016) and in the index finger (Z = -2.223, p= 0.026). The traditional scaling technique generated more pinch force (thumb x=7.25±4.99, index finger x=2.86±2.14) when compared to the modified scaling technique (thumb x=4.52±2.32, index finger x=1.65±1.28). Participants had a slightly higher preference for the instrument utilized for the modified scaling technique in terms of balance, maneuverability, overall comfort and the associated scaling technique as compared to the instrument utilized for the traditional scaling technique.Conclusion: Use of a modified scaling technique may reduce muscle activity and pinch force generation as compared to the traditional lateral pressure scaling technique during instrumentation. Future research on ergonomic scaling techniques is needed to determine their efficacy and impact on musculoskeletal disorders.


The Link Between Age and Partner Preferences in a Large, International Sample of Single Women.
Women's capacity to reproduce varies over the life span, and developmental goals such as family formation are age-graded and shaped by social norms about the appropriate age for completing specific developmental tasks. Thus, a woman's age may be linked to her ideas about what an ideal partner should be like. With the goals of replicating and extending prior research, in this study we examined the role of age in women's partner preferences across the globe. We investigated associations of age with ideal long-term partner preferences in a cross-cultural sample of 17,254 single (i.e., unpartnered) heterosexual women, ages 18 to 67, from 147 countries. Data were collected via an online questionnaire, the Ideal Partner Survey. Confirming our preregistered hypotheses, we found no or only negligible age effects on preferences for kindness-supportiveness, attractiveness, financial security-successfulness, or education-intelligence. Age was, however, positively associated with preferences for confidence-assertiveness. Consistent with family formation goals, age was associated with an ideal partner's parenting intentions (high until approximately age 30, then decreasing afterward). Age range deemed acceptable (and in particular, the discrepancy between one's own age and the minimum ideal age of a partner) increased with age. This latter pattern also replicated in exploratory analyses based on subsamples of lesbian and bisexual women. In summary, age has a limited impact on partner preferences. Of the attributes investigated, only preference for confidence-assertiveness was linked with age. However, age range deemed acceptable and an ideal partner's parenting intention, a dimension mostly neglected in earlier research, substantially vary with age.


Chemical rescue of Klebsiella aerogenes urease variants lacking the carbamylated-lysine nickel ligand.
Klebsiella aerogenes urease possesses a dinuclear metallocenter in which two nickel atoms are bridged by carbamylated Lys217. To assess whether carbamate-specific chemistry is required for urease activity, site-directed mutagenesis and chemical rescue strategies were combined in efforts to place a carboxylate group at the location of this metal ligand. Urease variants with Lys217 replaced by Glu, Cys, and Ala (K217E, K217C/C319A, and K217A proteins) were purified, shown to be activated by incubation with small organic acids plus Ni(II), and structurally characterized. K217C/C319A urease possessed a second change in which Cys319 was replaced by Ala in order to facilitate efforts to chemically modify Cys217; however, this covalent modification approach did not produce active urease. Chemical rescue of the K217E, K217C/C319A, and K217A variants required 2, 2, and 10 h, respectively, to reach maximal activity levels. The highest activity generated [224 micromol of urea degraded.min-1.(mg of protein)-1, for K217C/C319A urease incubated with 500 mM formic acid and 10 mM Ni at pH 6.5] corresponded to 56% of that measured for in vitro activation of the wild-type apoprotein. While the K217E apoprotein showed minimal structural perturbations, the K217C/C319A apoprotein showed a disordering of some active site residues, and the K217A apoprotein revealed a repositioning of His219 to allow the formation of a hydrogen bond with Thr169, thus replacing the hydrogen bond between the amino group of Lys217 and Thr169 in the native enzyme. Importantly, these structures allow rationalization of the relative rates and yields of chemical rescue experiments. The crystal structures of chemically rescued K217A and K217C/C319A ureases revealed a return of the active site residues to their wild-type positions. In both cases, noncovalently bound formate was structurally equivalent to the Lys-carbamate as the bridging metallocenter ligand. We conclude that carbamate-specific chemistry is not required for urease catalysis.


Soilwater Conductivity Analysis to Date and Locate Clandestine Graves of Homicide Victims.
In homicide investigations, it is critically important that postmortem interval and postburial interval (PBI) of buried victims are determined accurately. However, clandestine graves can be difficult to locate; and the detection rates for a variety of search methods (ranging from simple ground probing through to remote imaging and near-surface geophysics) can be very low. In this study, simulated graves of homicide victims were emplaced in three sites with contrasting soil types, bedrock, and depositional environments. The long-term monthly in situ monitoring of grave soil water revealed rapid increases in conductivity up to 2 years after burial, with the longest study evidencing declining values to background levels after 4.25 years. Results were corrected for site temperatures and rainfall to produce generic models of fluid conductivity as a function of time. The research suggests soilwater conductivity can give reliable PBI estimates for clandestine burials and therefore be used as a grave detection method.


Spatiotemporal Patterns of Adaptation-Induced Slow Oscillations in a Whole-Brain Model of Slow-Wave Sleep.
During slow-wave sleep, the brain is in a self-organized regime in which slow oscillations (SOs) between up- and down-states travel across the cortex. While an isolated piece of cortex can produce SOs, the brain-wide propagation of these oscillations are thought to be mediated by the long-range axonal connections. We address the mechanism of how SOs emerge and recruit large parts of the brain using a whole-brain model constructed from empirical connectivity data in which SOs are induced independently in each brain area by a local adaptation mechanism. Using an evolutionary optimization approach, good fits to human resting-state fMRI data and sleep EEG data are found at values of the adaptation strength close to a bifurcation where the model produces a balance between local and global SOs with realistic spatiotemporal statistics. Local oscillations are more frequent, last shorter, and have a lower amplitude. Global oscillations spread as waves of silence across the undirected brain graph, traveling from anterior to posterior regions. These traveling waves are caused by heterogeneities in the brain network in which the connection strengths between brain areas determine which areas transition to a down-state first, and thus initiate traveling waves across the cortex. Our results demonstrate the utility of whole-brain models for explaining the origin of large-scale cortical oscillations and how they are shaped by the connectome.


Multiplex PCR to diagnose bloodstream infections in patients admitted from the emergency department with sepsis.
Sepsis is caused by a heterogeneous group of infectious etiologies. Early diagnosis and the provision of appropriate antimicrobial therapy correlate with positive clinical outcomes. Current microbiological techniques are limited in their diagnostic capacities and timeliness. Multiplex PCR has the potential to rapidly identify bloodstream infections and fill this diagnostic gap. We identified patients from two large academic hospital emergency departments with suspected sepsis. The results of a multiplex PCR that could detect 25 bacterial and fungal pathogens were compared to those of blood culture. The results were analyzed with respect to the likelihood of infection, sepsis severity, the site of infection, and the effect of prior antibiotic therapy. We enrolled 306 subjects with suspected sepsis. Of these, 43 were later determined not to have infectious etiologies. Of the remaining 263 subjects, 70% had sepsis, 16% had severe sepsis, and 14% had septic shock. The majority had a definite infection (41.5%) or a probable infection (30.7%). Blood culture and PCR performed similarly with samples from patients with clinically defined infections (areas under the receiver operating characteristic curves, 0.64 and 0.60, respectively). However, blood culture identified more cases of septicemia than PCR among patients with an identified infectious etiology (66 and 46, respectively; P = 0.0004). The two tests performed similarly when the results were stratified by sepsis severity or infection site. Blood culture tended to detect infections more frequently among patients who had previously received antibiotics (P = 0.06). Conversely, PCR identified an additional 24 organisms that blood culture failed to detect. Real-time multiplex PCR has the potential to serve as an adjunct to conventional blood culture, adding diagnostic yield and shortening the time to pathogen identification.


Unexpected T-cell recognition of an altered peptide ligand is driven by reversed thermodynamics.
The molecular basis underlying T-cell recognition of MHC molecules presenting altered peptide ligands is still not well-established. A hierarchy of T-cell activation by MHC class I-restricted altered peptide ligands has been defined using the T-cell receptor P14 specific for H-2D(b) in complex with the immunodominant lymphocytic choriomeningitis virus peptide gp33 (KAVYNFATM). While substitution of tyrosine to phenylalanine (Y4F) or serine (Y4S) abolished recognition by P14, the TCR unexpectedly recognized H-2D(b) in complex with the alanine-substituted semiagonist Y4A, which displayed the most significant structural modification. The observed functional hierarchy gp33 > Y4A > Y4S = Y4F was neither due to higher stabilization capacity nor to differences in structural conformation. However, thermodynamic analysis demonstrated that while recognition of the full agonist H-2D(b) /gp33 was strictly enthalpy driven, recognition of the weak agonist H-2D(b) /Y4A was instead entropy driven with a large reduction in the favorable enthalpy term. The fourfold larger negative heat capacity derived for the interaction of P14 with H-2D(b) /gp33 compared with H-2D(b) /Y4A can possibly be explained by higher water entrapment at the TCR/MHC interface, which is also consistent with the measured opposite entropy contributions for the interactions of P14 with both MHCs. In conclusion, this study demonstrates that P14 makes use of different strategies to adapt to structural modifications in the MHC/peptide complex.


Marginal healing using Polyetheretherketone as healing abutments: an experimental study in dogs.
To evaluate the marginal soft and hard tissue healing at titanium and Polyetheretherketone (PEEK) healing implant abutments over a 4-month period. In six Labrador dogs, all mandibular premolars and first molars were extracted. After 4 months of healing, flaps were elevated, and two implants were installed at each side of the mandible, one in the premolar and the other in the molar regions. Four different types of healing abutments were positioned on the top of each implant: (i) titanium (Ti); (ii) PEEK material bonded to a base made of titanium (Ti-P), randomly positioned in the premolar region; (iii) PEEK, pristine (P); and (iv) PEEK, roughened (P-R), randomly positioned in the molar region. The flaps were sutured to allow a non-submerged healing, and after 4 months, the animals were sacrificed and ground sections obtained for histological evaluation. A higher resorption of the buccal bone crest was observed at the PEEK bonded to a base made of titanium abutments (1.0 ± 0.3 mm) compared to those made of titanium (0.3 ± 0.4 mm). However, similar dimensions of the peri-implant mucosa and similar locations of the soft tissues in relation to the implant shoulder were observed. No statistically significant differences were seen in the outcomes when the pristine PEEK was compared with the roughened PEEK abutments. The mean apical extension of the junctional epithelium did not exceed the implant shoulder at any of the abutment types used. The coronal level of the hard and soft tissues allows the conclusion that the use of PEEK as healing abutments may be indicated.


Role of triple test score in the evaluation of palpable breast lump.
Fine-needle aspiration cytology (FNAC) method is safe, reliable and time saving outdoor procedure with little discomfort to the patient for detecting Carcinoma breast. The efficacy can further be enhanced when physical breast examination, mammography and FNAC (the triple test [TT]) are jointly taken into consideration. The aim was to evaluate the role of TT score (TTS) in palpable breast masses. This prospective study was carried out from May 2010 to April 2012. In the subjects referred to the Department of Pathology for FNAC of the breast mass, the TTS was calculated, and histopathological findings were noted. In the study period TTS score was calculated in 200 cases out of 225 FNAC's of breast. Of 124 benign cases on cytology, only three showed discordant TTS. Out of 62 malignant cases, 61 showed concordant TTS and one case of mastitis on histopathology showed TTS of five. Out of all the benign lesions, two cases of fibrocystic disease and a single case of phylloides tumor gave a TTS ≥6. These cases were diagnosed as infiltrating ductal carcinoma and angiosarcoma respectively on histopathology. Histopathological correlation was possible in only 70 patients. Of these 70, 28 were from the benign category and 42 were from the malignant category. TTS of ≥6 has a sensitivity of 97.44%, specificity of 100%. FNAC has a sensitivity of 88.37%, specificity of 96.42%. TT reliably guides evaluation of palpable breast masses. Histological correlation indicated TTS to be better diagnostic tool than FNAC alone.


Enhanced cell adhesion to silicone implant material through plasma surface modification.
Silicone implant material is widely used in the field of plastic surgery. Despite its benefits the lack of biocompatibility this material still represents a major problem. Due to the surface characteristics of silicone, protein adsorption and cell adhesion on this polymeric material is rather low. The aim of this study was to create a stable collagen I surface coating on silicone implants via glow-discharge plasma treatment in order to enhance cell affinity and biocompatibility of the material. Non-plasma treated, collagen coated and conventional silicone samples (non-plasma treated, non-coated) served as controls. After plasma treatment the change of surface free energy was evaluated by drop-shape analysis. The quality of the collagen coating was analysed by electron microscopy and Time-Of-Flight Secondary Ion Mass Spectrometry. For biocompatibility tests mouse fibroblasts 3T3 were cultivated on the different silicone surfaces and stained with calcein-AM and propidium iodine to evaluate cell viability and adherence. Analysis of the different surfaces revealed a significant increase in surface free energy after plasma pre-treatment. As a consequence, collagen coating could only be achieved on the plasma activated silicone samples. The in vitro tests showed that the collagen coating led to a significant increase in cell adhesion and cell viability.


All-oral daclatasvir plus asunaprevir for hepatitis C virus genotype 1b: a multinational, phase 3, multicohort study.
An unmet need exists for interferon-free and ribavirin-free treatments for chronic hepatitis C virus (HCV) infection. In this study, we assessed all-oral therapy with daclatasvir (NS5A replication complex inhibitor) plus asunaprevir (NS3 protease inhibitor) in patients with genotype 1b infection, including those with high unmet needs or cirrhosis, or both. We did this phase 3, multicohort study (HALLMARK-DUAL) at 116 sites in 18 countries between May 11, 2012, and Oct 9, 2013. Patients were adults with chronic HCV genotype 1b infection who were treatment-naive; previous non-responders to peginterferon alfa plus ribavirin; or medically ineligible for, previously intolerant of, or ineligible for and intolerant of peginterferon alfa plus ribavirin. Treatment-naive patients were randomly assigned (2:1 ratio) by an interactive voice-response system with a computer-generated random allocation sequence (stratified by cirrhosis status) to receive daclatasvir 60 mg once daily plus asunaprevir 100 mg twice daily or placebo for 12 weeks. Patients and investigator sites were masked to treatment assignment and HCV RNA results to the end of week 12. The treatment-naive group assigned to daclatasvir plus asunaprevir continued open-label treatment to the end of week 24; participants assigned to placebo entered another daclatasvir plus asunaprevir study. Non-responders and ineligible, intolerant, or ineligible and intolerant patients received open-label daclatasvir plus asunaprevir for 24 weeks. The primary endpoint was sustained virological response at post-treatment week 12. Efficacy analyses were restricted to patients given daclatasvir plus asunaprevir. This trial is registered with ClinicalTrials.gov, number NCT01581203. This study included 307 treatment-naive patients (205 received daclatasvir plus asunaprevir and 102 received placebo; all randomly assigned patients received the intended treatment), 205 non-responders, and 235 ineligible, intolerant, or ineligible and intolerant patients. Daclatasvir plus asunaprevir provided sustained virological response in 182 (90%, 95% CI 85-94) patients in the treatment-naive cohort, 168 (82%, 77-87) in the non-responder cohort, and 192 (82%, 77-87) in the ineligible, intolerant, or ineligible and intolerant cohort. Serious adverse events occurred in 12 (6%) patients in the treatment-naive group; 11 (5%) non-responders, and 16 (7%) ineligible, intolerant, or ineligible and intolerant patients; adverse events leading to discontinuation (most commonly reversible increases in alanine or aspartate aminotransferase) occurred in six (3%), two (1%), and two (1%) patients, respectively, with no deaths recorded. Grade 3 or 4 laboratory abnormalities were uncommon, with low incidences of aminotransferase increases during the first 12 weeks with daclatasvir plus asunaprevir and placebo in treatment-naive patients (≤2% each). Daclatasvir plus asunaprevir provided high sustained virological response rates in treatment-naive, non-responder, and ineligible, intolerant, or ineligible and intolerant patients, and was well tolerated in patients with HCV genotype 1b infection. These results support the use of daclatasvir plus asunaprevir as an all-oral, interferon-free and ribavirin-free treatment option for patients with HCV genotype 1b infection, including those with cirrhosis. Bristol-Myers Squibb.


Influence of cytotoxic T lymphocyte antigen-4 (CTLA-4) gene polymorphisms in periodontitis.
Persistent host inflammatory immune response against the pathogens results in the destruction of periodontal tissues. Cytotoxic T lymphocyte antigen 4 (CTLA-4) is a particularly important molecule in down-regulating T-cell expansion and cytokine production. This study aimed to assess three functional SNPs within CTLA-4 gene, -1722 T/C, -318 C/T, and +49 A/G in patients with aggressive or chronic periodontitis. A total of 197 patients with periodontitis (71 aggressive and 126 chronic periodontitis) and 218 healthy controls were recruited. All samples were genotyped for CTLA-4 gene polymorphisms by polymerase chain reaction-amplification refractory mutation system (PCR-ARMS). The allelic and genotype frequencies of only +49 A/G SNP were more prominence in patients with chronic periodontitis (CP) than that controls (0.0005 and 0.001, respectively). Multivariate logistic regression analysis was demonstrated that homozygosity in +49 G/G had profoundly increased susceptibility for CP, OR=3.7 (95% CI; 1.6-8.5, P=0.001). In addition, comparison of CTLA-4 SNPs between patients with CP and aggressive periodontitis (AgP) revealed that heterozygosity in -1722 T/C polymorphism of CTLA-4 gene had a significantly higher risk for CP compared with AgP with a calculated odds ratio of 2.18 (95% CI; 1.17-4.06, P=0.01). These results suggest that CTLA-4 gene variants might be associated to susceptibility to specific form of periodontitis and participate in the CP development.


Validation and analysis of expression, prognosis and immune infiltration of WNT gene family in non-small cell lung cancer.
Early diagnosis and prognosis prediction of non-small cell lung cancer (NSCLC) have been challenging. Signaling cascades involving the Wingless-type (WNT) gene family play important biological roles and show prognostic value in various cancers, including NSCLC. On this basis, this study aimed to investigate the significance of WNTs in the prognosis and tumor immunity in NSCLC by comprehensive analysis. Expression and methylation levels of WNTs were obtained from the ONCOMINE, TIMER, and UALCAN. The dataset obtained from The Cancer Genome Atlas (TCGA) was utilized for prognostic analysis. cBioPortal was used to perform genetic alterations and correlation analysis of WNTs. R software was employed for functional enrichment and pathway analysis, partial statistics, and graph drawing. TRRUST was used to find key transcription factors. GEPIA was utilized for the analysis of expression, pathological staging, etc. Correlative analysis of immune infiltrates from TIMER. TISIDB was used for further immune infiltration validation analysis. Compared with that of normal tissues, WNT2/2B/3A/4/7A/9A/9B/11 expressions decreased, while WNT3/5B/6/7B/8B/10A/10B/16 expressions increased in lung adenocarcinoma (LUAD); WNT2/3A/7A/11 expressions were lessened, while WNT2B/3/5A/5B/6/7B/10A/10B/16 expressions were enhanced in squamous cell lung cancer (LUSC). Survival analysis revealed that highly expressed WNT2B and lowly expressed WNT7A predicted better prognostic outcomes in LUAD and LUSC. In the study of immune infiltration levels, WNT2, WNT9B, and WNT10A were positively correlated with six immune cells in LUAD; WNT1, WNT2, and WNT9B were positively correlated with six immune cells in LUSC, while WNT7B was negatively correlated. Our study indicated that WNT2B and WNT7A might have prognostic value in LUAD, and both of them might be important prognostic factors in LUSC and correlated to immune cell infiltration in LUAD and LUSC to a certain extent. Considering the prognostic value of WNT2B and WNT7A in NSCLC, we validated their mRNA and protein expression levels in NSCLC by performing qRT-PCR, western blot, and immunohistochemical staining on NSCLC pathological tissues and cell lines. This study may provide some direction for the subsequent exploration of the prognostic value of the WNTs and their role as biomarkers in NSCLC.


Recurrent Acute Pancreatitis: International State-of-the-Science Conference With Recommendations.
Recurrent acute pancreatitis (RAP) is a clinically significant problem globally. The etiology remains unclear in approximately 10% to 15% of patients despite a thorough workup. Data on natural history and efficacy of treatments are limited. We aimed to establish criteria for diagnosis, evaluate the causative factors, and arrive at a consensus on the appropriate workup and management of patients with RAP. The organizing committee was formed, and a set of questions was developed based on the current evidence, controversies, and topics that needed further research. After a vetting process, these topics were assigned to a group of experts from around the world with special interest in RAP. Data were presented as part of a workshop on RAP organized as a part of the annual meeting of the America Pancreatic Association. Pretest and Posttest questions were administered, and the responses were tabulated by the current Grades of Recommendation Assessment, Development and Evaluation system. The consensus guidelines were established in the format of a diagnostic algorithm. Several deficiencies were identified with respect to data on etiology, treatment efficacies, and areas that need immediate research.


Centenarians--a useful model for healthy aging? A 29-year follow-up of hospitalizations among 40,000 Danes born in 1905.
Centenarians surpass the current human life expectancy with about 20-25 years. However, whether centenarians represent healthy aging still remains an open question. Previous studies have been hampered by a number of methodological shortcomings such as a cross-sectional design and lack of an appropriate control group. In a longitudinal population-based cohort, it was examined whether the centenarian phenotype may be a useful model for healthy aging. The study was based on a complete follow up of 39,945 individuals alive in the Danish 1905 birth cohort on January 1, 1977 identified through the Danish Civil Registration System (DCRS). Data from the Danish Demographic Database and The Danish National Patient Register (in existence since 1977) were used. The 1905 cohort was followed up from 1977 through 2004 with respect to hospitalizations and number of hospital days. Survival status was available until December 2006. Danish centenarians from the 1905 cohort were hospitalized substantially less than their shorter-lived contemporaries at the same point in time during the years 1977 through 2004. For example, at age 71-74, the proportion of nonhospitalized centenarians was 80.5% compared with 68.4% among individuals who died in their early 80s. This trend was evident in both sexes. As a result of their lower hospitalization rates and length of stay in hospital compared with their contemporaries, who died at younger ages, Danish centenarians represent healthy agers. Centenarians constitute a useful study population in the search for fixed traits associated with exceptional longevity, such as genotype.


The impact of cannabis use on cognitive functioning in patients with schizophrenia: a meta-analysis of existing findings and new data in a first-episode sample.
Cannabis use is highly prevalent among people with schizophrenia, and coupled with impaired cognition, is thought to heighten the risk of illness onset. However, while heavy cannabis use has been associated with cognitive deficits in long-term users, studies among patients with schizophrenia have been contradictory. This article consists of 2 studies. In Study I, a meta-analysis of 10 studies comprising 572 patients with established schizophrenia (with and without comorbid cannabis use) was conducted. Patients with a history of cannabis use were found to have superior neuropsychological functioning. This finding was largely driven by studies that included patients with a lifetime history of cannabis use rather than current or recent use. In Study II, we examined the neuropsychological performance of 85 patients with first-episode psychosis (FEP) and 43 healthy nonusing controls. Relative to controls, FEP patients with a history of cannabis use (FEP + CANN; n = 59) displayed only selective neuropsychological impairments while those without a history (FEP - CANN; n = 26) displayed generalized deficits. When directly compared, FEP + CANN patients performed better on tests of visual memory, working memory, and executive functioning. Patients with early onset cannabis use had less neuropsychological impairment than patients with later onset use. Together, these findings suggest that patients with schizophrenia or FEP with a history of cannabis use have superior neuropsychological functioning compared with nonusing patients. This association between better cognitive performance and cannabis use in schizophrenia may be driven by a subgroup of "neurocognitively less impaired" patients, who only developed psychosis after a relatively early initiation into cannabis use.


CYP2C19 polymorphism and proton pump inhibitors.
Proton pump inhibitors such as omeprazole (esomeprazole), lansoprazole, pantoprazole and rabeprazole are eliminated by the hepatic route and the polymorphic CYP2C19 is mainly involved in their metabolism. In different populations three phenotypes have been identified: extensive metabolizers, poor metabolizers and individuals carrying one wild type and one mutant allele (het extensive metabolizers). Systemic exposure to the proton pump inhibitors as expressed by the AUC (area under the plasma level time profiles) is 5-12-times higher in poor metabolizers than in extensive metabolizers. As the pharmacodynamic response (elevation of intragastric pH) to the proton pump inhibitors is related directly to their AUC, a much higher pH can be monitored over 24 hr in poor metabolizers than in extensive metabolizers. Furthermore, clinical efficacy of all proton pump inhibitors depend on maintaining intragastric pH above certain threshold levels and significantly higher eradication rates of Helicobacter pylori have been observed in patients of the poor metabolizers and het extensive metabolizers phenotype if compared to extensive metabolizers. Likewise, limited data suggest that proton pump inhibitors-induced healing rates in gastro-oesophageal reflux disease are apparently higher in poor metabolizers/het extensive metabolizers than in extensive metabolizers of CYP2C19. Therefore initial genotyping for this enzyme and higher dosage in extensive metabolizers is likely to improve the clinical efficacy of proton pump inhibitors.


Learning endoscopic resection in the esophagus.
Endoscopic resection is the cornerstone of endoscopic management of esophageal early neoplasia. However, endoscopic resection is a complex technique requiring knowledge and expertise. Our aims were to identify the most important learning points in performing endoscopic resection in a training setting and to provide information on how to improve endoscopic resection technique. Six gastroenterologists at centers with multidisciplinary expertise in upper gastrointestinal oncology participated in a structured endoscopic resection training program, consisting of four training days with lectures and hands-on training on live pigs, further one-to-one hands-on training days, and written feedback (by an expert) on videos of unsupervised endoscopic resection procedures. The first 20 endoscopic resections of each participant were prospectively registered. Ninety learning points were independently identified by participants using a standardized questionnaire and by an expert providing written feedback on 33 unsupervised endoscopic resection videos. Three expert endoscopists selected and ranked the most important learning points in a consensus meeting. Results. The top 10 tips (illustrated by unique videos of three perforations) were: (1) allow time for inspection and use a high-definition endoscope; (2) create a preprocedural plan by placing electrocoagulation markings; (3) know the management of bleeding; (4) optimize the endoscopic view by repeatedly cleaning out stomach and target area; (5) use a therapeutic endoscope during resection; (6) always perform a test suction; (7) keep instruments close to the tip; (8) lift edges in piecemeal endoscopic cap resections; (9) know the management of perforation; (10) pin specimens down. This study summarized the most important learning points for performing endoscopic resection encountered during a structured endoscopic resection training program.


A nanopaper-based artificial tongue: a ratiometric fluorescent sensor array on bacterial nanocellulose for chemical discrimination applications.
In the present study, a ratiometric fluorescent sensor array as an artificial tongue has been developed on a nanopaper platform for chemical discrimination applications. The bacterial cellulose (BC) nanopaper was utilized for the first time as a novel, flexible, and transparent substrate in the optical sensor arrays for developing high-performance artificial tongues. To fabricate this platform, the hydrophobic walls on the BC nanopaper substrates were successfully created using a laser printing technology. In addition, we have used the interesting photoluminescence (PL) properties of an immobilized ratiometric probe (carbon dot-Rhodamine B (CD-RhB) nanohybrids) on the nanopaper platform to improve the visual discrimination analysis. Heavy metal ions were utilized as model analytes to verify the applicability of the fabricated nanopaper-based ratiometric fluorescent sensor array (NRFSA). Using the color variation of the NRFSA platform upon the addition of heavy metal ions, which have been obtained by a smartphone (under an UV irradiation), five heavy metal ions (i.e., Hg(ii), Pb(ii), Cd(ii), Fe(iii), and Cu(ii)) have been well-distinguished through the RGB analysis via production of the characteristic PL fingerprint-like response patterns for each of them. Moreover, the developed optical sensor array was successfully exploited to identify the heavy metal ions in the water and fish samples. We have also found that the PL spectra, which have been obtained by a spectrofluorometer, of the developed NRFSA can be exploited for discrimination applications. We believe that the nanopaper-based artificial tongues will provide innovative insights into the development of optical sensor arrays towards advanced (bio)chemical discrimination applications and can revolutionize the conventional optical sensor array technology.


The dynamic continence challenge - a simple test to predict faecal continence prior to colostomy closure.
A common problem in clinical practice is predicting whether a patient will be continent after treatment of a severe perineal injury. Several tests have been described. Anal manometry is unreliable; continence can be normal with low pressures, and poor with high or normal pressures. Endo-anal ultrasound only illustrates anatomical sphincter integrity. The saline continence test involves the quite unphysiological instilling of saline into the rectum, and assessing seepage. What is needed in the prediction of continence is a normal stool simulator. We propose the use of powdered instant mashed potato reconstituted with water to the consistency of faeces. About 100 - 150 ml is introduced into the rectum using a catheter-tipped syringe. The patient is instructed to walk around for half an hour. On return the underwear is examined for any soiling. If there is no leakage the colostomy may be reversed. Over the past 15 years, 53 patients have undergone this test. In 47 patients there was no leakage, all had their stomas reversed, and none was incontinent during follow-up. The dynamic continence challenge is an accurate physiological test that allows clinicians to simulate the effects of colostomy reversal and assess a patient's continence before actually proceeding to the reversal.


Pathology in euthermic bats with white nose syndrome suggests a natural manifestation of immune reconstitution inflammatory syndrome.
White nose syndrome, caused by Geomyces destructans, has killed more than 5 million cave hibernating bats in eastern North America. During hibernation, the lack of inflammatory cell recruitment at the site of fungal infection and erosion is consistent with a temperature-induced inhibition of immune cell trafficking. This immune suppression allows G. destructans to colonize and erode the skin of wings, ears and muzzle of bat hosts unchecked. Yet, paradoxically, within weeks of emergence from hibernation an intense neutrophilic inflammatory response to G. destructans is generated, causing severe pathology that can contribute to death. We hypothesize that the sudden reversal of immune suppression in bats upon the return to euthermia leads to a form of immune reconstitution inflammatory syndrome (IRIS). IRIS was first described in HIV-infected humans with low helper T lymphocyte counts and bacterial or fungal opportunistic infections. IRIS is a paradoxical and rapid worsening of symptoms in immune compromised humans upon restoration of immunity in the face of an ongoing infectious process. In humans with HIV, the restoration of adaptive immunity following suppression of HIV replication with anti-retroviral therapy (ART) can trigger severe immune-mediated tissue damage that can result in death. We propose that the sudden restoration of immune responses in bats infected with G. destructans results in an IRIS-like dysregulated immune response that causes the post-emergent pathology.


Archipelago-wide island restoration in the Galápagos Islands: reducing costs of invasive mammal eradication programs and reinvasion risk.
Invasive alien mammals are the major driver of biodiversity loss and ecosystem degradation on islands. Over the past three decades, invasive mammal eradication from islands has become one of society's most powerful tools for preventing extinction of insular endemics and restoring insular ecosystems. As practitioners tackle larger islands for restoration, three factors will heavily influence success and outcomes: the degree of local support, the ability to mitigate for non-target impacts, and the ability to eradicate non-native species more cost-effectively. Investments in removing invasive species, however, must be weighed against the risk of reintroduction. One way to reduce reintroduction risks is to eradicate the target invasive species from an entire archipelago, and thus eliminate readily available sources. We illustrate the costs and benefits of this approach with the efforts to remove invasive goats from the Galápagos Islands. Project Isabela, the world's largest island restoration effort to date, removed >140,000 goats from >500,000 ha for a cost of US$10.5 million. Leveraging the capacity built during Project Isabela, and given that goat reintroductions have been common over the past decade, we implemented an archipelago-wide goat eradication strategy. Feral goats remain on three islands in the archipelago, and removal efforts are underway. Efforts on the Galápagos Islands demonstrate that for some species, island size is no longer the limiting factor with respect to eradication. Rather, bureaucratic processes, financing, political will, and stakeholder approval appear to be the new challenges. Eradication efforts have delivered a suite of biodiversity benefits that are in the process of revealing themselves. The costs of rectifying intentional reintroductions are high in terms of financial and human resources. Reducing the archipelago-wide goat density to low levels is a technical approach to reducing reintroduction risk in the short-term, and is being complemented with a longer-term social approach focused on education and governance.


The mechanism of local tumor irradiation combined with interleukin 2 therapy in murine renal carcinoma: histological evaluation of pulmonary metastases.
We have demonstrated that tumor irradiation enhanced the therapeutic effect of interleukin 2 (IL-2) on pulmonary metastases from a murine renal adenocarcinoma, Renca. To investigate the mechanism of interaction between tumor irradiation and IL-2 therapy, we have histologically evaluated the effects of each therapy alone or in combination on Renca pulmonary metastases. Following treatment of established lung metastases with irradiation and IL-2 therapy, lung sections were processed for H&E or immunohistochemical staining. We found that tumor irradiation or IL-2 therapy locally induced vascular damage, resulting in multifocal hemorrhages and mononuclear cell mobilization in the lung tissue. This effect was amplified in lungs treated with the combined therapy. Immunohistochemistry showed that irradiation produced a macrophage influx into irradiated tumor nodules, and systemic IL-2 therapy induced T-cell infiltration in tumor nodules. Lungs treated with the combined therapy exhibited massive macrophage, T-cell, and natural killer cell mobilization in disintegrating tumor nodules and in the lung tissue. This combined therapy caused a decrease in the number of proliferating tumor cells and an increase in the number of apoptotic cells, which were more marked than with either therapy alone. We suggest that the macrophages mobilized by radiation-induced tissue injury could play a role in phagocytosis of apoptotic tumor cells, processing and presenting of tumor antigens for a systemic immune response activated by IL-2. Tumor destruction may result from the concomitant action of activated T cells, natural killer cells, and macrophages infiltrating the tumor nodules.


Infectious disease rates in the U.S. Navy, 1980 to 1995.
The effect of increasing numbers of women in the U.S. Navy, particularly those aboard Navy ships, on infectious disease risk is unknown. This study examines gender and other demographic differences among all U.S. Navy enlisted personnel in first hospitalizations for infectious diseases from 1980 through 1989 and identifies trends in incidence rates during the extended period from 1980 to 1995. All data were obtained from official personnel and medical records. First hospitalization rates were computed using the Epidemiological Interactive System. Varicella and other viruses and chlamydiae accounted for more than 20,000 hospitalizations among Navy enlisted personnel in the 1980s. In 7 of the 12 categories of common infectious diseases, women's rates were higher than those for men, particularly for viral meningitis, herpes simplex, syphilis, gonococcal disease, and candidiasis. An excess of certain common infectious diseases among women and nonwhite ethnic groups emphasizes the need for continuing education and surveillance in these populations.


Current modulation and membrane targeting of the calcium channel alpha1C subunit are independent functions of the beta subunit.
1. The beta subunits of voltage-sensitive calcium channels facilitate the incorporation of channels into the plasma membrane and modulate calcium currents. In order to determine whether these two effects of the beta subunit are interdependent or independent of each other we studied plasma membrane incorporation of the channel subunits with green fluorescent protein and immunofluorescence labelling, and current modulation with whole-cell and single-channel patch-clamp recordings in transiently transfected human embryonic kidney tsA201 cells. 2. Coexpression of rabbit cardiac muscle alpha1C with rabbit skeletal muscle beta1a, rabbit heart/brain beta2a or rat brain beta3 subunits resulted in the colocalization of alpha1C with beta and in a marked translocation of the channel complexes into the plasma membrane. In parallel, the whole-cell current density and single-channel open probability were increased. Furthermore, the beta2a isoform specifically altered the voltage dependence of current activation and the inactivation kinetics. 3. A single amino acid substitution in the beta subunit interaction domain of alpha1C (alpha1CY467S) disrupted the colocalization and plasma membrane targeting of both subunits without affecting the beta subunit-induced modulation of whole-cell currents and single-channel properties. 4. These results show that the modulation of calcium currents by beta subunits can be explained by beta subunit-induced changes of single-channel properties, but the formation of stable alpha1C-beta complexes and their increased incorporation into the plasma membrane appear not to be necessary for functional modulation.


A review on yolk sac utilization in poultry.
During incubation, embryonic growth and development are dependent on nutrients deposited in the egg. The content of the yolk can be transferred to the embryo in 2 ways: directly into the intestine via the yolk stalk or through the highly vascularized yolk sac membrane. It has been suggested that, as a result of genetic selection and improved management, the increase in posthatch growth rate and concurrently the increase in metabolic rate of broiler chickens during the last 50 yr has also increased embryonic metabolism. A higher metabolic rate during incubation would imply a lower residual yolk weight and possibly lower energy reserve for the hatchling. This might affect posthatch development and performance. This review examined scientific publications published between 1930 and 2018 to compare residual yolk weight at hatch, metabolic heat production, and yolk utilization throughout incubation. This review aimed to investigate 1) whether or not residual yolk weight and composition has been changed during the 88-yr period considered and 2) which abiotic and biotic factors affect yolk utilization in poultry during incubation and the early posthatch period. It can be concluded that 1) residual yolk weight and the total solid amount of the residual yolk at hatch seem to be decreased in the recent decades. It cannot be concluded whether the (lack of) differences between old and modern strains are due to genetic selection, changed management and incubation conditions, or moment of sampling (immediately after hatch or at pulling). It is remarkable that with the genetic progress and improved management and incubation conditions over the last 88 yr, effects on yolk utilization efficiency and embryonic metabolic heat production are limited; 2) factors specially affecting residual yolk weight at hatch include egg size and incubation temperature, whereas breeder age has more influence on nutrient composition of the residual yolk.


The LION Procedure to the Pelvic Nerves for Recovery of Locomotion in 18 Spinal Cord Injured Peoples - A Case Series.
We report on unexpected findings in 18 spinal cord injured peoples who underwent a laparoscopic bilateral implantation of neuroprosthesis (LION procedure) to the sciatic/femoral nerves pelvic somatic nerves for functional electrical stimulation (FES)-assisted locomotor training and continuous low-frequency electrical stimulation. Fifteen patients were paraplegics, three low tetraplegics, all of them fully dependent on a wheelchair. After a training period of at least one year, all patients not only started with electrical-assisted standing/stepping using a walker or crutches, but also developed some progressive caudalward recovery of lumbosacral sensoric functions and of supraspinal control of voluntary movements below the lesions. Twelve patients are currently capable of weight-bearing standing and stepping with crutches by simultaneous electrical stimulation (the best of the series for 2.6km), and six of them are capable of walking several meters (the best of the series for 400m) without electrical stimulation. Our findings suggest that FES-assisted locomotor training in combination with continuous low-frequency pelvic nerves in spinal cord injury patients may induce changes that affect the central pattern generator and allows supra- and infraspinal inputs to engage residual spinal or extra spinal pathways for reconnection.


Impact of intraoperative MRI-guided transsphenoidal surgery on endocrine function and hormone substitution therapy in patients with pituitary adenoma.
Pituitary adenomas are rare with an incidence of 0.4-8.2 per 105 inhabitants. Symptoms range from headaches to pituitary insufficiency or excessive output of hormones with associated disease. Except for prolactinomas, surgery is recommended as the first line and most effective treatment for the majority of these tumours. One of the refinements of surgical therapy introduced was intraoperative magnetic resonance imaging (iMRI). The aim of this study was to analyse the postoperative pituitary function and the general outcome of patients treated for non-functioning and GH-producing pituitary adenomas with a transsphenoidal iMRI-assisted approach using the PoleStar™ N20 imager. A total of 148 consecutive iMRI-guided surgeries for GH-producing and non-functioning pituitary adenomas were retrospectively analysed. Patients' clinical data, endocrinological parameters, clinical examinations and pre-/post- and intraoperative imaging studies were evaluated. A total of 101 patients could be classified as being in remission at follow-up; 26 (17.6%) of them due to iMRI allowing additional tumour removal. A total of 44 patients (29.7%) had more complete tumour removal because remnants were detected by iMRI. The mean hormone levels of patients did not differ significantly between pre- and postoperative examinations. There were 62 patients with preoperative, and 43 patients with postoperative pituitary insufficiency, thus, due to surgery there were 19 (12.8%) patients with improved pituitary function. The results show this method to be a safe and effective treatment option increasing remission rate and keeping complication rate low. Postoperative pituitary function was preserved or improved - possibly due to more exact iMRI-assisted tumour removal.


Quercetin effectively quells peanut-induced anaphylactic reactions in the peanut sensitized rats.
Peanut allergy is the major leading cause of fatal or life-threatening anaphylactic reactions to foods. At present, there is no remedy for this condition. The applied pharmaceutical cares are merely palliative, while their deleterious side effects have already been established. Hence, many sufferers search for complementary and alternative medicines. A versatile-, "flavonol" subgroup-member of the flavonoid family, quercetin, is of paramount interest to investigators. In this study the effects of quercetin on peanut-induced anaphylactic reactions were investigated in a rat model of peanut allergy. Wistar rats were sensitized with crude peanut extract in the presence of Cholera toxin and Aluminium hydroxide. Sensitized rats were then allotted into three groups; Positive control, Quercetin-treatment and Sham, (n=7, each). Naive rats (n=7) served as negative controls. One week post-sensitization period, the rats in treatment group were treated with quercetin at a dose of 50 mg/kg(Body Weight)/mL Di-methyl-sulfoxide 5%/rat, over a period of four weeks. Subsequently, rats were challenged, and anaphylactic reaction parameters including variations in plasma histamine levels, vascular permeability, systemic anaphylaxis scores, and total serum Immunoglobulin E levels were measured. After daily-gavaging for four weeks, quercetin completely abrogated peanut-induced anaphylactic reactions following challenges, so that the mean of plasma histamine levels in the quercetin-treated rats, were lower significantly (p=0.004) as compared with positive control group. Our findings suggest that the flavonoid quercetin is potent enough to suppress the on-going Immunoglobulin E responses against peanut proteins, and can be propounded as an alternative medicine to protect against Immunoglobulin E-mediated food allergies.


Prevalence and characteristics of functional dyspepsia among workers exposed to cement dust.
No data are currently available on the prevalence and characteristics of functional dyspepsia among cement workers. Given the potential impact of dyspepsia on work performance, whether its prevalence is increased among workers exposed to cement dust was evaluated. Altogether 420 cement workers were enrolled in the study. According to the individual exposure levels to cement dust, the following three groups were established: no exposure (128 workers), low exposure (1 mg/m3, 176 workers). Dyspepsia was evaluated by means of validated questionnaires. The prevalence of dyspepsia was increased in both the low and high exposure groups in comparison with the unexposed workers (51.7% and 59.1%, respectively, versus 34.4%; adjusted odds ratio (aOR) 2.21, 95% confidence interval (95% CI) 1.25-3.92, and aOR 2.36, 95% CI 1.31-4.25, respectively). A stepwise regression analysis showed a progressive increase in the odds ratio for dyspepsia for the low- and high-exposure groups with the no-exposure group as reference (OR 1.94, 95% CI 1.15-3.27, and OR 2.61, 95% CI 1.62-4.20, respectively). Ulcer-like dyspepsia was especially associated with the degree of exposure to cement dust, 11.71% for the no-exposure group versus 17.24% for the low-exposure group versus 29.54% for the high-exposure group (aOR 3.49, 95% CI 1.60-7.63), when the high-exposure group was compared with the no-exposure group. Similar findings were obtained for reflux-like dyspepsia. Occupational cement-dust exposure is associated with dyspepsia, and the association is particularly strong for ulcer-like and reflux-like dyspepsia.


CLR01 protects dopaminergic neurons in vitro and in mouse models of Parkinson's disease.
Parkinson's disease (PD) affects millions of patients worldwide and is characterized by alpha-synuclein aggregation in dopamine neurons. Molecular tweezers have shown high potential as anti-aggregation agents targeting positively charged residues of proteins undergoing amyloidogenic processes. Here we report that the molecular tweezer CLR01 decreased aggregation and toxicity in induced pluripotent stem cell-derived dopaminergic cultures treated with PD brain protein extracts. In microfluidic devices CLR01 reduced alpha-synuclein aggregation in cell somas when axonal terminals were exposed to alpha-synuclein oligomers. We then tested CLR01 in vivo in a humanized alpha-synuclein overexpressing mouse model; mice treated at 12 months of age when motor defects are mild exhibited an improvement in motor defects and a decreased oligomeric alpha-synuclein burden. Finally, CLR01 reduced alpha-synuclein-associated pathology in mice injected with alpha-synuclein aggregates into the striatum or substantia nigra. Taken together, these results highlight CLR01 as a disease-modifying therapy for PD and support further clinical investigation.


Racial/ethnic variation in parent expectations for antibiotics: implications for public health campaigns.
Widespread overuse and inappropriate use of antibiotics are a major public health concern. Little is known about racial/ethnic differences in parents seeking antibiotics for their children's upper respiratory illnesses. To examine racial/ethnic differences in parent expectations about the need for antibiotics and physician perceptions of those expectations. We conducted a nested, cross-sectional survey of parents who were coming to see their child's pediatrician because of cold symptoms between October 2000 and June 2001. Parents completed a previsit survey that collected information on demographics, their child's illness, and a 15-item previsit expectations inventory that included an item asking how necessary it was for the physician to prescribe antibiotics. Physicians completed a postvisit survey that collected information on diagnosis, treatment, and whether the physician perceived the parent expected an antibiotic. The encounter was the unit of analysis. Multivariate logistic regression analyses were performed to evaluate predictors of dichotomized parental expectations for antibiotics, dichotomized physician perceptions of those expectations, diagnostic patterns, and antibiotic-prescribing patterns. Twenty-seven community pediatric practices in the Los Angeles, Calif, metropolitan area. A volunteer sample of 38 pediatricians (participation rate: 64%) and a consecutive sample of 543 parents (participation rate: 83%; approximately 15 participating for each enrolled pediatrician) seeking care for their children's respiratory illnesses. Pediatricians were eligible to participate if they worked in a community-based managed care practice in the Los Angeles area. Parents were eligible to participate if they could speak and read English and presented to participating pediatricians with a child 6 months to 10 years old who had cold symptoms but had not received antibiotics within 2 weeks. Parental beliefs about the necessity of antibiotics for their child's illness, physician perceptions of parental expectations for antibiotics, bacterial diagnosis rates, and antibiotic-prescribing rates. Forty-three percent of parents believed that antibiotics were definitely necessary, and 27% believed that they were probably necessary for their child's illness. Latino and Asian parents were both 17% more likely to report that antibiotics were either definitely or probably necessary than non-Hispanic white parents. Physicians correctly perceived that Asian parents expected antibiotics more often than non-Hispanic white parents but underestimated the greater expectations of Latino parents for antibiotics. Physicians also correctly perceived that parents of children with ear pain or who were very worried about their child's condition were significantly more likely to expect antibiotics. Physicians were 7% more likely to make a bacterial diagnosis and 21% more likely to prescribe antibiotics when they perceived that antibiotics were expected. Parent expectations for antibiotics remain high in Los Angeles County. With time, traditional public health messages related to antibiotic use may decrease expectations among non-Hispanic white parents. However, both public health campaigns and physician educational efforts may need to be designed differently to reach other racial/ethnic groups effectively. Despite public health campaigns to reduce antibiotic overprescribing in the pediatric outpatient setting, physicians continue to respond to parental pressure to prescribe them. To effectively intervene to decrease rates of inappropriate antibiotic prescribing further, physicians need culturally appropriate tools to better communicate and negotiate with parents when feeling pressured to prescribe antibiotics.


A red-shifted photochromic sulfonylurea for the remote control of pancreatic beta cell function.
Azobenzene photoresponsive elements can be installed on sulfonylureas, yielding optical control over pancreatic beta cell function and insulin release. An obstacle to such photopharmacological approaches remains the use of ultraviolet-blue illumination. Herein, we synthesize and test a novel yellow light-activated sulfonylurea based on a heterocyclic azobenzene bearing a push-pull system.


Transformation optics for cavity array metamaterials.
Cavity array metamaterials (CAMs), composed of optical microcavities in a lattice coupled via tight-binding interactions, represent a novel architecture for engineering metamaterials. Since the size of the CAMs' constituent elements are commensurate with the operating wavelength of the device, it cannot directly utilise classical transformation optics in the same way as traditional metamaterials. By directly transforming the internal geometry of the system, and locally tuning the permittivity between cavities, we provide an alternative framework suitable for tight-binding implementations of metamaterials. We develop a CAM-based cloak as the case study.


Retinoic acid modulation of glutathione and cysteine metabolism in chondrocytes.
The major objective of this investigation was to determine the thiol status of chondrocytes and to relate changes in the level of glutathione and cysteine to maturation of the cells as they undergo terminal differentiation. Chondrocytes were isolated from the cephalic portion of chick embryo sterna and treated with all-trans retinoic acid for one week. We found that the addition of 100 nM retinoic acid to the cultures decreased the intracellular levels of glutathione and cysteine from 6.1 to 1.6 and 0.07 to 0.01 nmol/microgram DNA respectively; retinoic acid also caused a decrease in the extracellular concentration of cysteine. The decrease in chondrocyte thiols was dose and time dependent. To characterize other antioxidant systems of the sternal cell culture, the activities of catalase, glutathione reductase and superoxide dismutase were determined. Activities of all of those enzymes were high in the retinoic acid-treated cells; the conditioned medium also contained these enzymes and the cytosolic isoenzyme of superoxide dismutase. We probed the specificity of the thiol response by using immature caudal chondrocytes. Unlike the cephalic cells, retinoic acid did not change intracellular glutathione and extracellular cysteine levels, although the retinoid caused a reduction in the intracellular cysteine concentration. Finally, we explored the effect of medium components on chondrocyte thiol status. We noted that while ascorbate alone did not change cell thiol levels, it did cause a 4-fold decrease in the extracellular cysteine concentration. When retinoic acid and ascorbic acid were both present in the medium, there was a marked decrease in the level of glutathione. In contrast, the phosphate concentration of the culture medium served as a powerful modulator of both glutathione and cysteine. Results of the study clearly showed that there is a profound decrease in intracellular levels of both cysteine and glutathione and that thiol levels are responsive to ascorbic acid and the medium phosphate concentration. These findings point to a critical role for thiols in modulating events linked to chondrocyte maturation and cartilage matrix synthesis and mineralization.


Regulation of Mitogen-Activated Protein Kinase Signaling Pathways by the Ubiquitin-Proteasome System and Its Pharmacological Potential.
Mitogen-activated protein kinase (MAPK) cascades are evolutionarily conserved signaling pathways that play essential roles in transducing extracellular environmental signals into diverse cellular responses to maintain homeostasis. These pathways are classically organized into an architecture of three sequentially acting protein kinases: a MAPK kinase kinase that phosphorylates and activates a MAPK kinase, which in turn phosphorylates and activates the effector MAPK. The activity of MAPKs is tightly regulated by phosphorylation of their activation loop, which can be modulated by positive and negative feedback mechanisms to control the amplitude and duration of the signal. The signaling outcomes of MAPK pathways are further regulated by interactions of MAPKs with scaffolding and regulatory proteins. Accumulating evidence indicates that, in addition to these mechanisms, MAPK signaling is commonly regulated by ubiquitin-proteasome system (UPS)-mediated control of the stability and abundance of MAPK pathway components. Notably, the biologic activity of some MAPKs appears to be regulated mainly at the level of protein turnover. Recent studies have started to explore the potential of targeted protein degradation as a powerful strategy to investigate the biologic functions of individual MAPK pathway components and as a new therapeutic approach to overcome resistance to current small-molecule kinase inhibitors. Here, we comprehensively review the mechanisms, physiologic importance, and pharmacological potential of UPS-mediated protein degradation in the control of MAPK signaling. SIGNIFICANCE STATEMENT: Accumulating evidence highlights the importance of targeted protein degradation by the ubiquitin-proteasome system in regulating and fine-tuning the signaling output of mitogen-activated protein kinase (MAPK) pathways. Manipulating protein levels of MAPK cascade components may provide a novel approach for the development of selective pharmacological tools and therapeutics.


Investigation of different group A immunoassays following one dose of meningococcal group A conjugate vaccine or A/C polysaccharide vaccine in adults.
A double-blind, randomized, controlled phase I study to assess the safety, immunogenicity, and antibody persistence of a new group A conjugate vaccine (PsA-TT) in volunteers aged 18 to 35 years was previously performed. Subjects received one dose of either the PsA-TT conjugate vaccine, meningococcal A/C polysaccharide vaccine (PsA/C), or tetanus toxoid vaccine. The conjugate vaccine was shown to be safe and immunogenic as demonstrated by a standardized group A-specific immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and by a serum bactericidal antibody (SBA) assay using rabbit complement (rSBA). This report details further analysis of the sera using four additional immunologic assays to investigate the relationship between the different immunoassays. The immunoassays used were an SBA assay that used human complement (hSBA), a group A-specific IgG multiplexed bead assay, and two opsonophagocytic antibody (OPA) assays which used two different methodologies. For each vaccine group, geometric mean concentrations or geometric mean titers were determined for all assays before and 4, 24, and 48 weeks after vaccination. Pearson's correlation coefficients were used to assess the relationship between the six assays using data from all available visits. An excellent correlation was observed between the group A-specific IgG concentrations obtained by ELISA and those obtained by the multiplexed bead assay. hSBA and rSBA titers correlated moderately, although proportions of subjects with putatively protective titers and those demonstrating a > or = 4-fold rise were similar. The two OPA methods correlated weakly and achieved only a low correlation with the other immunoassays. The correlation between hSBA and group A-specific IgG was higher for the PsA-TT group than for the PsA/C group.


Challenges of Big Data Analysis.
Big Data bring new opportunities to modern society and challenges to data scientists. On one hand, Big Data hold great promises for discovering subtle population patterns and heterogeneities that are not possible with small-scale data. On the other hand, the massive sample size and high dimensionality of Big Data introduce unique computational and statistical challenges, including scalability and storage bottleneck, noise accumulation, spurious correlation, incidental endogeneity, and measurement errors. These challenges are distinguished and require new computational and statistical paradigm. This article gives overviews on the salient features of Big Data and how these features impact on paradigm change on statistical and computational methods as well as computing architectures. We also provide various new perspectives on the Big Data analysis and computation. In particular, we emphasize on the viability of the sparsest solution in high-confidence set and point out that exogeneous assumptions in most statistical methods for Big Data can not be validated due to incidental endogeneity. They can lead to wrong statistical inferences and consequently wrong scientific conclusions.


Different strokes for different folks: the rich diversity of animal models of focal cerebral ischemia.
No single animal model is able to encompass all of the variables known to affect human ischemic stroke. This review highlights the major strengths and weaknesses of the most commonly used animal models of acute ischemic stroke in the context of matching model and experimental aim. Particular emphasis is placed on the relationships between outcome and underlying vascular variability, physiologic control, and use of models of comorbidity. The aim is to provide, for novice and expert alike, an overview of the key controllable determinants of experimental stroke outcome to help ensure the most effective application of animal models to translational research.


GRIP: A web-based system for constructing Gold Standard datasets for protein-protein interaction prediction.
Information about protein interaction networks is fundamental to understanding protein function and cellular processes. Interaction patterns among proteins can suggest new drug targets and aid in the design of new therapeutic interventions. Efforts have been made to map interactions on a proteomic-wide scale using both experimental and computational techniques. Reference datasets that contain known interacting proteins (positive cases) and non-interacting proteins (negative cases) are essential to support computational prediction and validation of protein-protein interactions. Information on known interacting and non interacting proteins are usually stored within databases. Extraction of these data can be both complex and time consuming. Although, the automatic construction of reference datasets for classification is a useful resource for researchers no public resource currently exists to perform this task. GRIP (Gold Reference dataset constructor from Information on Protein complexes) is a web-based system that provides researchers with the functionality to create reference datasets for protein-protein interaction prediction in Saccharomyces cerevisiae. Both positive and negative cases for a reference dataset can be extracted, organised and downloaded by the user. GRIP also provides an upload facility whereby users can submit proteins to determine protein complex membership. A search facility is provided where a user can search for protein complex information in Saccharomyces cerevisiae. GRIP is developed to retrieve information on protein complex, cellular localisation, and physical and genetic interactions in Saccharomyces cerevisiae. Manual construction of reference datasets can be a time consuming process requiring programming knowledge. GRIP simplifies and speeds up this process by allowing users to automatically construct reference datasets. GRIP is free to access at http://rosalind.infj.ulst.ac.uk/GRIP/.


Catch and measure-mass spectrometry-based immunoassays in biomarker research.
Mass spectrometry-based (MS) methods are effective tools for discovering protein biomarker candidates that can differentiate between physiological and pathophysiological states. Promising candidates are validated in studies comprising large patient cohorts. Here, targeted protein analytics are used to increase sample throughput. Methods involving antibodies, such as sandwich immunoassays or Western blots, are commonly applied at this stage. Highly-specific and sensitive mass spectrometry-based immunoassays that have been established in recent years offer a suitable alternative to sandwich immunoassays for quantifying proteins. Mass Spectrometric ImmunoAssays (MSIA) and Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA/iMALDI) are two prominent types of MS-based immunoassays in which the capture is done either at the protein or the peptide level. We present an overview of these emerging types of immunoassays and discuss their suitability for the discovery and validation of protein biomarkers. This article is part of a Special Issue entitled: Biomarkers: A Proteomic Challenge.