Determinations of the peroxidative susceptibilities of cod liver oils by a newly-developed 1H NMR-based method: resistance of an antioxidant-fortified product isolated from pre-fermented sources.
To explore the molecular composition and antioxidant status of four natural (unrefined) cod liver oil (CLO) products, three of which (Products 1-3) were non-fermented, whilst one (Product 4) was isolated from pre-fermented cod livers, and hence was naturally antioxidant-fortified. Potential antioxidants and aldehyde-scavenging agents were determined by recommended and/or 1H NMR methods; peroxyl radical-specific oxygen radical absorbance capacity (ORAC) values were measured fluorimetrically. The activities of such antioxidants were also investigated by assessing the susceptibilities/resistivities of these CLOs to thermo-oxidation by 1H NMR analysis, which monitored the time-dependent evolution of aldehydic lipid oxidation products at 180 °C. Product 4 displayed much higher, albeit variable ORAC values (mean ± SEM 91.4 ± 19.5 mmol. trolox equivalents/kg) than those of Products 1-3, an observation arising from significant levels of peroxidation-blocking and/or aldehyde-consuming collagenous polypeptides/peptides, flavonones, biogenic amines, total phenolics, tannins, and ammoniacal agents therein. All of these agents were undetectable in Products 1-3. Quantitative considerations indicated that collagenous gel agents (present at ca. 1.5% (w/w)) were the most powerful Product 4 antioxidants. Significantly lower levels of aldehydes were generated in this product when exposed to thermal-stressing episodes. Results confirmed the enhanced peroxidative resistivity of a pre-fermented, antioxidant-rich natural CLO over those of corresponding non-fermented products. Product 4: Green Pasture Blue Ice™ fermented cod liver oil.


Evaluating Neanderthal genetics and phylogeny.
The retrieval of Neanderthal (Homo neanderthalsensis) mitochondrial DNA is thought to be among the most significant ancient DNA contributions to date, allowing conflicting hypotheses on modern human (Homo sapiens) evolution to be tested directly. Recently, however, both the authenticity of the Neanderthal sequences and their phylogenetic position outside contemporary human diversity have been questioned. Using Bayesian inference and the largest dataset to date, we find strong support for a monophyletic Neanderthal clade outside the diversity of contemporary humans, in agreement with the expectations of the Out-of-Africa replacement model of modern human origin. From average pairwise sequence differences, we obtain support for claims that the first published Neanderthal sequence may include errors due to postmortem damage in the template molecules for PCR. In contrast, we find that recent results implying that the Neanderthal sequences are products of PCR artifacts are not well supported, suffering from inadequate experimental design and a presumably high percentage (>68%) of chimeric sequences due to "jumping PCR" events.


Circulating tumor DNA in advanced prostate cancer: transitioning from discovery to a clinically implemented test.
The genomic landscape of metastatic castration-resistant prostate cancer (mCRPC) differs from that of the primary tumor and is dynamic during tumor progression. The real-time and repeated characterization of this process via conventional solid tumor biopsies is challenging. Alternatively, circulating cell-free DNA (cfDNA) containing circulating tumor DNA (ctDNA) can be obtained from patient plasma using minimally disruptive blood draws and is amenable to sequential analysis. ctDNA has high overlap with the genomic sequences of biopsies from metastases and has the advantage of being representative of multiple metastases. The availability of techniques with high sensitivity and specificity, such as next-generation sequencing (NGS) and digital PCR, has greatly contributed to the development of the cfDNA field and enabled the detection of genomic alterations at low ctDNA fractions. In mCRPC, a number of clinically relevant genomic alterations have been tracked in ctDNA, including androgen receptor (AR) aberrations, which have been shown to be associated with an adverse outcome to novel antiandrogen therapies, and alterations in homologous recombination repair (HRR) genes, which have been associated with a response to PARP inhibitors. Several clinical applications have been proposed for cfDNA analysis, including its use as a prognostic tool, as a predictive biomarker, to monitor tumor response and to identify novel mechanisms of resistance. To date, the cfDNA analysis has provided interesting results, but there is an urgent need for these findings to be confirmed in prospective clinical trials.


Association between pelvic muscle mass and canine hip dysplasia.
To investigate the relationship between pelvic muscle mass and development and expression of canine hip dysplasia (CHD). Prospective study. 5 Greyhounds with anatomically normal hip joints, 59 German Shepherd Dogs (23 with CHD, 24 with near-normal hip joints, and 12 with normal hip joints), and 18 German Shepherd Dog-Greyhound crossbreeds (7 with CHD, 6 with near-normal hip joints, and 5 with normal hip joints) between 12 and 47 months old in which pelvic muscle mass was evaluated. Pectineal muscle and hip joint development were evaluated in 25 German Shepherd Dogs at 8 and 16 or 24 weeks of age. For evaluation of pelvic muscle mass, individual pelvic muscles were weighed and hip joints were assigned a score on the basis of severity of degenerative changes. For evaluation of pectineal muscle development, muscle sections were stained and examined. Pelvic muscle mass was greatest in Greyhounds, intermediate in crossbred dogs, and smallest in German Shepherd Dogs. Differences in pelvic muscle mass among breeds were attributable to differences in weights of individual muscles. Hip score was negatively correlated with pelvic muscle mass and weights of selected pelvic muscles. Dogs with pectineal hypotrophy at 8 weeks of age had type-2 muscle fiber paucity or muscle fiber-type grouping at 16 or 24 weeks of age. At 8 weeks of age, hip joints were composed of multiple centers of ossification, and the acetabulum was largely cartilaginous. By 24 weeks of age, the pelvic bones were largely, although incompletely, fused. Diminished pelvic muscle mass in dogs with CHD and altered muscle fiber size and composition in 8-week-old dogs that subsequently develop CHD strongly suggest that abnormalities of pelvic musculature are associated with development of CHD. The complex development of the hip joint from multiple centers of ossification may make the joint susceptible to abnormal modeling forces that would result from abnormalities in pelvic muscle mass.


Early detection of diabetic kidney disease by urinary proteomics and subsequent intervention with spironolactone to delay progression (PRIORITY): a prospective observational study and embedded randomised placebo-controlled trial.
Microalbuminuria is an early sign of kidney disease in people with diabetes and indicates increased risk of cardiovascular disease. We tested whether a urinary proteomic risk classifier (CKD273) score was associated with development of microalbuminuria and whether progression to microalbuminuria could be prevented with the mineralocorticoid receptor antagonist spironolactone. In this multicentre, prospective, observational study with embedded randomised controlled trial (PRIORITY), we recruited people with type 2 diabetes, normal urinary albumin excretion, and preserved renal function from 15 specialist centres in ten European countries. All participants (observational cohort) were tested with the CKD273 classifier and classified as high risk (CKD273 classifier score >0·154) or low risk (≤0·154). Participants who were classified as high risk were entered into a randomised controlled trial and randomly assigned (1:1), by use of an interactive web-response system, to receive spironolactone 25 mg once daily or matched placebo (trial cohort). The primary endpoint was development of confirmed microalbuminuria in all individuals with available data (observational cohort). Secondary endpoints included reduction in incidence of microalbuminuria with spironolactone (trial cohort, intention-to-treat population) and association between CKD273 risk score and measures of impaired renal function based on estimated glomerular filtration rate (eGFR; observational cohort). Adverse events (particularly gynaecomastia and hyperkalaemia) and serious adverse events were recorded for the intention-to-treat population (trial cohort). This study is registered with the EU Clinical Trials Register (EudraCT 20120-004523-4) and ClinicalTrials.gov (NCT02040441) and is completed. Between March 25, 2014, and Sept 30, 2018, we enrolled and followed-up 1775 participants (observational cohort), 1559 (88%) of 1775 participants had a low-risk urinary proteomic pattern and 216 (12%) had a high-risk pattern, of whom 209 were included in the trial cohort and assigned to spironolactone (n=102) or placebo (n=107). The overall median follow-up time was 2·51 years (IQR 2·0-3·0). Progression to microalbuminuria was seen in 61 (28%) of 216 high-risk participants and 139 (9%) of 1559 low-risk participants (hazard ratio [HR] 2·48, 95% CI 1·80-3·42; p<0·0001, after adjustment for baseline variables of age, sex, HbA1c, systolic blood pressure, retinopathy, urine albumin-to-creatinine ratio [UACR], and eGFR). Development of impaired renal function (eGFR <60 mL/min per 1·73 m2) was seen in 48 (26%) of 184 high-risk participants and 119 (8%) of 1423 low-risk participants (HR 3·50; 95% CI 2·50-4·90, after adjustment for baseline variables). A 30% decrease in eGFR from baseline (post-hoc endpoint) was seen in 42 (19%) of 216 high-risk participants and 62 (4%) of 1559 low-risk participants (HR 5·15, 95% CI 3·41-7·76; p<0·0001, after adjustment for basline eGFR and UACR). In the intention-to-treat trial cohort, development of microalbuminuria was seen in 35 (33%) of 107 in the placebo group and 26 (25%) of 102 in the spironolactone group (HR 0·81, 95% CI 0·49-1·34; p=0·41). In the safety analysis (intention-to-treat trial cohort), events of plasma potassium concentrations of more than 5·5 mmol/L were seen in 13 (13%) of 102 participants in the spironolactone group and four (4%) of 107 participants in the placebo group, and gynaecomastia was seen in three (3%) participants in the spironolactone group and none in the placebo group. One patient died in the placebo group due to a cardiac event (considered possibly related to study drug) and one patient died in the spironolactone group due to cancer, deemed unrelated to study drug. In people with type 2 diabetes and normoalbuminuria, a high-risk score from the urinary proteomic classifier CKD273 was associated with an increased risk of progression to microalbuminuria over a median of 2·5 years, independent of clinical characteristics. However, spironolactone did not prevent progression to microalbuminuria in high-risk patients. European Union Seventh Framework Programme.


Toxoplasma gondii Infection and Mixed Anxiety and Depressive Disorder: A Case-Control Seroprevalence Study in Durango, Mexico.
The parasite Toxoplasma gondii (T. gondii) may invade the brain and might induce behavioral changes. We sought to determine the association of T. gondii infection and mixed anxiety and depressive disorder. Through an age- and gender-matched case-control seroprevalence study, we examined 65 patients suffering from mixed anxiety and depressive disorder (WHO ICD-10 code: F41.2) attending in a public hospital of mental health and 260 control subjects without this disorder from the general population. Sera of participants were analyzed for anti-Toxoplasma IgG and IgM antibodies using enzyme-linked immunoassays. Fifteen (23.1%) of the 65 patients and 18 (6.9%) of the 260 controls had anti-T. gondii IgG antibodies (odds ratio (OR): 4.03; 95% confidence interval (CI): 1.90 - 8.53; P < 0.001). The frequency of high (> 150 IU/mL) anti-T. gondii IgG levels was similar in cases and controls (OR: 0.25; 95% CI: 0.05 - 1.06; P = 0.05). Seroprevalence was similar in male cases and controls (P = 1.0); however, seroprevalence was significantly higher in female cases than in female controls (OR: 7.08; 95% CI: 2.83 - 17.67; P < 0.00001). Patients aged 31 - 50 years old had a significantly higher seroprevalence of T. gondii infection than controls of the same age group (OR: 21.04; 95% CI: 5.22 - 84.80; P < 0.00001). Anti-T. gondii IgM antibodies were found in four (26.7%) of the 15 anti-T. gondii IgG seropositive cases and in 10 (55.6%) of the 18 anti-T. gondii IgG seropositive controls (P = 0.15). Results support for the first time an association between seropositivity to T. gondii and mixed anxiety and depressive disorder. Further research to confirm this association and to determine the seroepidemiology of T. gondii infection in patients with this disorder is needed.


Thrombospondin-1 is induced in rat myocardial infarction and its induction is accelerated by ischemia/reperfusion.
Thrombospondin-1 (TSP-1) is a multifunctional, rapid-turnover matricellular protein. Recent studies demonstrated that TSP-1 has a role in regulating inflammatory reactions. Myocardial infarction (MI) is associated with an inflammatory response, ultimately leading to healing and scar formation. In particular, an enhanced inflammatory reaction and a massive accumulation of monocytes/macrophages is seen with reperfusion after MI. To examine the role of TSP-1 in MI, we isolated rat TSP-1 complementary DNA (cDNA) and analyzed the level and distribution of the mRNA expression. In infarcted rat hearts, TSP-1 mRNA increased markedly at 6 and 12 hrs after coronary artery ligation (27.97 +/- 3.40-fold and 22.77 +/- 1.83-fold, respectively, compared with sham-operated hearts). Western blot analysis revealed that TSP-1 protein was transiently induced in the infarcted heart. Using in situ hybridization analysis, TSP-1 mRNA signals were observed in the infiltrating cells at the border area of infarction. We then examined the effect of ischemia/reperfusion (I/R) on TSP-1 mRNA induction in the rats with infarcted hearts. Quantitative reverse transcriptase polymerase chain reaction (RT-PCR) demonstrated that I/R enhanced the TSP-1 mRNA expression approximately 4-fold, as compared with the level in the permanently ligated heart. Finally, we examined the effect of TSP-1 on proinflammatory cytokine release in mononuclear cells. The releases of interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) from human mononuclear cells were enhanced by TSP-1 in a dose-dependent manner. Thus, the immediate and marked increase of TSP-1 expression suggests that TSP-1 has an inflammatory-associated role in MI.


Phase-sensitive four-wave mixing interferometer.
A nonlinear interferometer is constructed using a dual-pump, dual-signal four-wave mixing (FWM) process. The generated idlers are categorized into phase-insensitive and phase-sensitive (PS) outputs according to whether the power is sensitive to the phase difference of the input signals. The PS idlers are obtained from the interference of the primary idlers and are regarded as the FWM interferometer output. Potential applications include photonic exclusive-NOR and exclusive-OR operations, photonic correlators, and photonic microwave filters.


Ergodic and nonergodic many-body dynamics in strongly nonlinear lattices.
The study of nonlinear oscillator chains in classical many-body dynamics has a storied history going back to the seminal work of Fermi et al. [Los Alamos Scientific Laboratory Report No. LA-1940, 1955 (unpublished)]. We introduce a family of such systems which consist of chains of N harmonically coupled particles with the nonlinearity introduced by confining the motion of each individual particle to a box or stadium with hard walls. The stadia are arranged on a one-dimensional lattice but they individually do not have to be one dimensional, thus permitting the introduction of chaos already at the lattice scale. For the most part we study the case where the motion is entirely one dimensional. We find that the system exhibits a mixed phase space for any finite value of N. Computations of Lyapunov spectra at randomly picked phase space locations and a direct comparison between Hamiltonian evolution and phase space averages indicate that the regular regions of phase space are not significant at large system sizes. While the continuum limit of our model is itself a singular limit of the integrable sinh Gordon theory, we do not see any evidence for the kind of nonergodicity famously seen in the work of Fermi et al. Finally, we examine the chain with particles confined to two-dimensional stadia where the individual stadium is already chaotic and find a much more chaotic phase space at small system sizes.


Cardiac arrest survival is rare without prehospital return of spontaneous circulation.
Emergency medical services (EMS) are crucial in the management of out-of-hospital cardiac arrest (OHCA). Despite accepted termination-of-resuscitation criteria, many patients are transported to the hospital without achieving field return of spontaneous circulation (ROSC). We examine field ROSC influence on OHCA survival to hospital discharge in two large urban EMS systems. A retrospective analysis of prospectively collected data was conducted. Data collection is a component of San Antonio Fire Department's comprehensive quality assurance/quality improvement program and Cincinnati Fire Department's participation in the Cardiac Arrest Registry to Enhance Survival (CARES) project. Attempted resuscitations of medical OHCA and cardiac OHCA for San Antonio and Cincinnati, respectively, from 2008 to 2010 were analyzed by city and in aggregate. A total of 2,483 resuscitation attempts were evaluated. Age and gender distributions were similar between cities, but ethnic profiles differed. Cincinnati had 17% (p = 0.002) more patients with an initial shockable rhythm and was more likely to initiate transport before field ROSC. Overall survival to hospital discharge was 165 of 2,483 (6.6%). More than one-third (894 of 2,483, 36%) achieved field ROSC. Survival with field ROSC was 17.2% (154 of 894) and without field ROSC was 0.69% (11 of 1,589). Of the 11 survivors transported prior to field ROSC, nine received defibrillation by EMS. No asystolic patient survived to hospital discharge without field ROSC. Survival to hospital discharge after OHCA is rare without field ROSC. Resuscitation efforts should focus on achieving field ROSC. Transport should be reserved for patients with field ROSC or a shockable rhythm.


'Your country needs you': the ethics of allocating staff to high-risk clinical roles in the management of patients with COVID-19.
As the COVID-19 pandemic impacts on health service delivery, health providers are modifying care pathways and staffing models in ways that require health professionals to be reallocated to work in critical care settings. Many of the roles that staff are being allocated to in the intensive care unit and emergency department pose additional risks to themselves, and new policies for staff reallocation are causing distress and uncertainty to the professionals concerned. In this paper, we analyse a range of ethical issues associated with changes to staff allocation processes in the face of COVID-19. In line with a dominant view in the medical ethics literature, we claim, first, that no individual health professional has a specific, positive obligation to treat a patient when doing so places that professional at risk of harm, and so there is a clear ethical tension in any reallocation process in this context. Next, we argue that the changing asymmetries of health needs in hospitals means that careful consideration needs to be given to a stepwise process for deallocating staff from their usual duties. We conclude by considering how a justifiable process of reallocating professionals to high-risk clinical roles should be configured once those who are 'fit for reallocation' have been identified. We claim that this process needs to attend to three questions that we consider in detail: (1) how the choice to make reallocation decisions is made, (2) what justifiable models for reallocation might look like and (3) what is owed to those who are reallocated.


Imaging and sizing of diamond nanoparticles.
Typical disturbances of biological environment such as background scatter and refractive index variations have little effect on the size-dependent scattering property of highly refractive nanocrystals, which are potentially attractive optical labels. We report on what is to our knowledge the first investigation of these scattering optical labels, and their sizing, in particular, by imaging at subvideo frame rates and analyzing samples of diamond nanocrystals deposited on a glass substrate in air and in a matrix of weakly scattering polymer. The brightness of a diffraction-limited spot appears to serve as a reliable measure of the particle size in the Rayleigh scattering limit.


Human cadaveric study of the morphology of the basilar artery.
Nourishment for the brain, a highly vascular organ, is derived from a unique structure called the 'circle of Willis', which is formed by the terminal branches of the internal carotid arteries (ICAs) and basilar arteries (BAs). The circle of Willis forms an anastomotic link between the carotid and vertebrobasilar systems in the arterial supply of the brain, while the BA forms an important component of the brain's posterior circulation and supplies its many vital parts. A study was performed on 20 brain specimens used for routine dissections at the Anatomy Department, Kasturba Medical College, in order to examine the morphology of BAs in the brain. In most specimens, the position of the termination of BA was normal, although variations were present in the mode of termination. In one specimen, the BA terminated by dividing into two superior cerebellar arteries. The posterior cerebral arteries (PCAs) arose from ICAs on both sides in this specimen, and a communicating branch was present between the terminal point of the BA and PCA on the left. In another specimen, unilateral variation was seen, with the PCA arising from the ICA on the right and a posterior communicating artery arising from the PCA, connecting it with the BA. The anatomy on the left side was normal. We highlight the morphological aspects of the BA, the knowledge of which would help neurosurgeons safely diagnose, as well as plan and execute vascular bypass and shunting procedures for the treatment of stenosis, aneurysms and arteriovenous malformations in the posterior cranial fossa.


Medical Care Costs Associated with Cancer Survivorship in the United States.
The prevalence of cancer survivorship is increasing. In this study, we provide contemporary population-based estimates and projections of the overall and site-specific cancer-attributable medical care costs in the United States. We identified survivors aged ≥65 years diagnosed with cancer between 2000 and 2012 from the Surveillance, Epidemiology, and End Results (SEER)-Medicare database and used 2007 to 2013 claims to estimate costs by cancer site, phases of care, and stage at diagnosis. Annualized average cancer-attributable costs for medical care (Medicare Parts A and B) and oral prescription drugs (Medicare Part D) were estimated by subtracting costs between patients with cancer and matched controls. Costs are reported in 2019 U.S. dollars. We combined phase-specific attributable costs with prevalence projections to estimate national costs from 2015 through 2030. Overall annualized average costs were highest in the end-of-life-cancer death phase, followed by the initial and continuing phases (medical care: $105,500, $41,800, and $5,300 and oral prescription drugs: $4,200, $1,800, $1,100, respectively). There was considerable variation in costs by cancer site and stage. Overall national costs in 2015 were $183 billion and projected to increase 34% to $246 billion by 2030, based only on population growth. Phase of care cancer-attributable cost estimates by cancer site and stage are key inputs for simulation models and cost-effectiveness analyses. The national cancer-attributed medical care costs in the United States are substantial and projected to increase dramatically by 2030, due to population changes alone, reflecting the rising burden of cancer care among cancer survivors.


Price to be paid for two-metal catalysis: magnesium ions that accelerate chemistry unavoidably limit product release from a protein kinase.
Incorporation of divalent metal ions into an active site is a fundamental catalytic tool used by diverse enzymes. Divalent cations are used by protein kinases to both stabilize ATP binding and accelerate chemistry. Kinetic analysis establishes that Cyclin-dependent kinase 2 (CDK2) requires simultaneous binding of two Mg(2+) ions for catalysis of phosphoryl transfer. This tool, however, comes with a price: the rate-acceleration effects are opposed by an unavoidable rate-limiting consequence of the use of two Mg(2+) ions by CDK2. The essential metal ions stabilize ADP product binding and limit the overall rate of the reaction. We demonstrate that product release is rate limiting for activated CDK2 and evaluate the effects of the two catalytically essential Mg(2+) ions on the stability of the ADP product within the active site. We present two new crystal structures of CDK2 bound to ADP showing how the phosphate groups can be coordinated by either one or two Mg(2+) ions, with the occupancy of one site in a weaker equilibrium. Molecular dynamics simulations indicate that ADP phosphate mobility is more restricted when ADP is coordinated by two Mg(2+) ions compared to one. The structural similarity between the rigid ADP·2Mg product and the cooperatively assembled transition state provides a mechanistic rational for the rate-limiting ADP release that is observed. We demonstrate that although the simultaneous binding of two Mg(2+) ions is essential for efficient phosphoryl transfer, the presence of both Mg(2+) ions in the active site also cooperatively increases ADP affinity and opposes its release. Evolution of protein kinases must have involved careful tuning of the affinity for the second Mg(2+) ion in order to balance the needs to stabilize the chemical transition state and allow timely product release. The link between Mg(2+) site affinity and activity presents a chemical handle that may be used by regulatory factors as well as explain some mutational effects.


Determination of the complex refractive indices of aerosol from aerodynamic particle size spectrometer and integrating nephelometer measurements.
An approach is developed to retrieve the complex index of refraction by coupling two well known instruments: an aerodynamic particle size spectrometer (APS) probe for measuring aerosol size distributions and an integrating nephelometer for measuring total light scattering coefficients. The retrieval is realized by an iterative least squares minimization of the fractional error between the nephelometer-measured light scattering coefficients and those calculated from the APS-measured size distributions based on the Mie theory for spherical particles. High-resolution data collected during two field experiments conducted at two locations with distinct environments in China are analyzed. The results show that light scattering coefficients, aerosol size distributions, and refractive indices all vary substantially with time. Further examination of their dependence on relative humidity suggests that instead of being monotonic change with relative humidity, the refractive index often fluctuates when the relative humidity changes. This nonmonotonic variation of refractive index with relative index suggests concurrent change of relative humidity and other chemical compositions. Possible errors in the retrieval are also discussed.


Quality and quantity: mucosal CD4+ T cells and HIV susceptibility.
The HIV sexual transmission probability measured in the context of discordant couples appears too low to fuel the HIV pandemic, but these rates are substantially amplified by specific co-factors. The most consistent predictors of transmission are the HIV levels in the blood and genital tract of an infected individual, each of which increases the transmission probability in a dose-dependent manner. In an analogous fashion, we propose that both the quantity and quality of HIV-susceptible target cells in the exposed genital or rectal mucosa may be key predictors of HIV susceptibility. The absolute number of mucosal CD4+ T cells is increased in several situations that are associated with amplified HIV transmission, particularly during genital infections. In addition, qualitative mucosal T-cell parameters such as immune activation and the expression of the HIV binding molecules CCR5 and/or α4β7 are important determinants of gp120 binding and productive HIV infection. In particular, the Th17 and Th22 cell subsets are enhanced within mucosal compartments and appear to be highly HIV-susceptible. Blockade of specific HIV target cell subsets at the site of exposure, if done in a safe and effective manner, represents an opportunity for new HIV prevention tools.


Common α-globin variants modify hematologic and other clinical phenotypes in sickle cell trait and disease.
Co-inheritance of α-thalassemia has a significant protective effect on the severity of complications of sickle cell disease (SCD), including stroke. However, little information exists on the association and interactions for the common African ancestral α-thalassemia mutation (-α3.7 deletion) and β-globin traits (HbS trait [SCT] and HbC trait) on important clinical phenotypes such as red blood cell parameters, anemia, and chronic kidney disease (CKD). In a community-based cohort of 2,916 African Americans from the Jackson Heart Study, we confirmed the expected associations between SCT, HbC trait, and the -α3.7 deletion with lower mean corpuscular volume/mean corpuscular hemoglobin and higher red blood cell count and red cell distribution width. In addition to the recently recognized association of SCT with lower estimated glomerular filtration rate and glycated hemoglobin (HbA1c), we observed a novel association of the -α3.7 deletion with higher HbA1c levels. Co-inheritance of each additional copy of the -α3.7 deletion significantly lowered the risk of anemia and chronic kidney disease among individuals with SCT (P-interaction = 0.031 and 0.019, respectively). Furthermore, co-inheritance of a novel α-globin regulatory variant was associated with normalization of red cell parameters in individuals with the -α3.7 deletion and significantly negated the protective effect of α-thalassemia on stroke in 1,139 patients with sickle cell anemia from the Cooperative Study of Sickle Cell Disease (CSSCD) (P-interaction = 0.0049). Functional assays determined that rs11865131, located in the major alpha-globin enhancer MCS-R2, was the most likely causal variant. These findings suggest that common α- and β-globin variants interact to influence hematologic and clinical phenotypes in African Americans, with potential implications for risk-stratification and counseling of individuals with SCD and SCT.


Angiography for assessment of preoperative chemotherapy in musculoskeletal sarcomas.
Eleven abnormal findings of digital subtraction angiography were analyzed in 25 patients with bone sarcoma and in 23 patients with soft tissue sarcoma. The relation between digital subtraction angiographic findings and the histologic effect of chemotherapy was evaluated. Digital subtraction angiography was performed with the patient under local anesthesia with the Seldinger technique, using an ADVANT X unit. Contrast medium was injected at a rate of 4 to 6 ml per second, with the usual single bolus dose of approximately 6 to 8 ml. Eleven abnormal findings included tumor stain, hypervascularity, arterial distortion, vascular stretch, arterial dilatation, arteriovenous shunt, arterial encasement, occlusion, blood pool, caliber with irregular wall, and dilatation of draining vein. Each finding after preoperative chemotherapy was compared with that before chemotherapy and divided into three grades; Grade 1, not effective; Grade 2, effective; and Grade 3, very effective. Angiographic Grades 2 and 3 were defined as responders. The histologic effect was examined and scored according to the modified classification proposed by Huvos. Histologic Grades 1 and 2 were classified as nonresponders and Grades 3 and 4 as good responders. In bone sarcomas, hypervascularity and tumor stain were seen in all patients. In soft tissue sarcomas, tumor stain was shown in all 23 patients and hypervascularity was seen in 21 patients. Tumor stain, hypervascularity, vascular stretch, and arterial encasement were correlated closely with histologic findings and showed an accuracy equal to or greater than 70%. When these four findings changed to angiographic Grade 3 after preoperative chemotherapy, 90% of patients had good histologic outcome.


Dynamical predictors of an imminent phenotypic switch in bacteria.
Single cells can stochastically switch across thresholds imposed by regulatory networks. Such thresholds can act as a tipping point, drastically changing global phenotypic states. In ecology and economics, imminent transitions across such tipping points can be predicted using dynamical early warning indicators. A typical example is 'flickering' of a fast variable, predicting a longer-lasting switch from a low to a high state or vice versa. Considering the different timescales between metabolite and protein fluctuations in bacteria, we hypothesized that metabolic early warning indicators predict imminent transitions across a network threshold caused by enzyme saturation. We used stochastic simulations to determine if flickering predicts phenotypic transitions, accounting for a variety of molecular physiological parameters, including enzyme affinity, burstiness of enzyme gene expression, homeostatic feedback, and rates of metabolic precursor influx. In most cases, we found that metabolic flickering rates are robustly peaked near the enzyme saturation threshold. The degree of fluctuation was amplified by product inhibition of the enzyme. We conclude that sensitivity to flickering in fast variables may be a possible natural or synthetic strategy to prepare physiological states for an imminent transition.


Predicting Adverse Outcomes for Shiga Toxin-Producing Escherichia coli Infections in Emergency Departments.
To assess the performance of a hemolytic uremic syndrome (HUS) severity score among children with Shiga toxin-producing Escherichia coli (STEC) infections and HUS by stratifying them according to their risk of adverse events. The score has not been previously evaluated in a North American acute care setting. We reviewed medical records of children <18 years old infected with STEC and treated in 1 of 38 participating emergency departments in North America between 2011 and 2015. The HUS severity score (hemoglobin [g/dL] plus 2-times serum creatinine [mg/dL]) was calculated using first available laboratory results. Children with scores >13 were designated as high-risk. We assessed score performance to predict severe adverse events (ie, dialysis, neurologic complication, respiratory failure, and death) using discrimination and net benefit (ie, threshold probability), with subgroup analyses by age and day-of-illness. A total of 167 children had HUS, of whom 92.8% (155/167) had relevant data to calculate the score; 60.6% (94/155) experienced a severe adverse event. Discrimination was acceptable overall (area under the curve 0.71, 95% CI 0.63-0.79) and better among children <5 years old (area under the curve 0.77, 95% CI 0.68-0.87). For children <5 years, greatest net benefit was achieved for a threshold probability >26%. The HUS severity score was able to discriminate between high- and low-risk children <5 years old with STEC-associated HUS at a statistically acceptable level; however, it did not appear to provide clinical benefit at a meaningful risk threshold.


Hypoxia inducible factor-alpha binding and ubiquitylation by the von Hippel-Lindau tumor suppressor protein.
The von Hippel-Lindau tumor suppressor protein (pVHL) has emerged as a key factor in cellular responses to oxygen availability, being required for the oxygen-dependent proteolysis of alpha subunits of hypoxia inducible factor-1 (HIF). Mutations in VHL cause a hereditary cancer syndrome associated with dysregulated angiogenesis, and up-regulation of hypoxia inducible genes. Here we investigate the mechanisms underlying these processes and show that extracts from VHL-deficient renal carcinoma cells have a defect in HIF-alpha ubiquitylation activity which is complemented by exogenous pVHL. This defect was specific for HIF-alpha among a range of substrates tested. Furthermore, HIF-alpha subunits were the only pVHL-associated proteasomal substrates identified by comparison of metabolically labeled anti-pVHL immunoprecipitates from proteosomally inhibited cells and normal cells. Analysis of pVHL/HIF-alpha interactions defined short sequences of conserved residues within the internal transactivation domains of HIF-alpha molecules sufficient for recognition by pVHL. In contrast, while full-length pVHL and the p19 variant interact with HIF-alpha, the association was abrogated by further N-terminal and C-terminal truncations. The interaction was also disrupted by tumor-associated mutations in the beta-domain of pVHL and loss of interaction was associated with defective HIF-alpha ubiquitylation and regulation, defining a mechanism by which these mutations generate a constitutively hypoxic pattern of gene expression promoting angiogenesis. The findings indicate that pVHL regulates HIF-alpha proteolysis by acting as the recognition component of a ubiquitin ligase complex, and support a model in which its beta domain interacts with short recognition sequences in HIF-alpha subunits.


Avalanche prediction in a self-organized pile of beads.
It is a common belief that power-law distributed avalanches are inherently unpredictable. This idea affects phenomena as diverse as evolution, earthquakes, superconducting vortices, stock markets, etc., from atomic to social scales. It mainly comes from the concept of "self-organized criticality" (SOC), where criticality is interpreted in the way that, at any moment, any small avalanche can eventually cascade into a large event. Nevertheless, this work demonstrates experimentally the possibility of avalanche prediction in the classical paradigm of SOC: a pile of grains. By knowing the position of every grain in a two-dimensional pile, avalanches of moving grains follow a distinct power-law distribution. Large avalanches, although uncorrelated, are on average preceded by continuous, detectable variations in the internal structure of the pile that are monitored in order to achieve prediction.


Apolipoprotein A1 as a novel urinary biomarker for diagnosis of bladder cancer: A systematic review and meta-analysis.
The emergence of urinary biomarkers for bladder cancer diagnosis could provide a reliable and less invasive diagnostic method. It could be also used as an adjuvant to the current gold standards of cytology and cystoscopy to improve diagnostic accuracy and decrease the percentage of false positives. We searched PubMed, SCOPUS, and Web of Science up to March 18, 2020. We selected four studies that assessed the diagnostic accuracy of urinary apolipoprotein A1 (ApoA-1) in detecting bladder cancer and met the inclusion and exclusion criteria. Two authors independently extracted the data and performed quality assessment of the studies. Four studies with 771 participants were selected; 417 were bladder cancer patients and 354 were controls. Bladder cancer was either transitional cell carcinoma or squamous cell carcinoma, the stages varied between Ta to T3, and the grades varied between G1 and G3. The pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio were 90.7%, 90%, 9.478, 0.1, and 99.424, respectively. Summary receiver operating characteristic curve showed an area under the curve of 0.9544 and Q* index of 0.8965. ApoA-1 showed high sensitivity and specificity, so it could be a useful biomarker in diagnosis of bladder cancer.


N-O-isopropyl sulfonamido-based hydroxamates: design, synthesis and biological evaluation of selective matrix metalloproteinase-13 inhibitors as potential therapeutic agents for osteoarthritis.
Matrix metalloproteinase-13 (MMP-13) is a key enzyme implicated in the degradation of the extracellular matrix in osteoarthritis (OA). For this reason, MMP-13 synthetic inhibitors are being sought as potential therapeutic agents to prevent cartilage degradation and to halt the progression of OA. Herein, we report the synthesis and in vitro evaluation of a new series of selective MMP-13 inhibitors possessing an arylsulfonamidic scaffold. Among these potential inhibitors, a very promising compound was discovered exhibiting nanomolar activity for MMP-13 and was highly selective for this enzyme compared to MMP-1, -14, and TACE. This compound acted as a slow-binding inhibitor of MMP-13 and was demonstrated to be effective in an in vitro collagen assay and in a model of cartilage degradation. Furthermore, a docking study was conducted for this compound in order to investigate its binding interactions with MMP-13 and the reasons for its selectivity toward MMP-13 versus other MMPs.


The transcriptome of Plasmodium vivax reveals divergence and diversity of transcriptional regulation in malaria parasites.
Plasmodium vivax causes over 100 million clinical infections each year. Primarily because of the lack of a suitable culture system, our understanding of the biology of this parasite lags significantly behind that of the more deadly species P. falciparum. Here, we present the complete transcriptional profile throughout the 48-h intraerythrocytic cycle of three distinct P. vivax isolates. This approach identifies strain specific patterns of expression for subsets of genes predicted to encode proteins associated with virulence and host pathogen interactions. Comparison to P. falciparum revealed significant differences in the expression of genes involved in crucial cellular functions that underpin the biological differences between the two parasite species. These data provide insights into the biology of P. vivax and constitute an important resource for the development of therapeutic approaches.


Modification of intracellular free calcium in cultured F2408 embryo fibroblasts by 3-substituted-2-thiohydantoin derivatives.
3-substituted-2-thiohydantoin derivatives were synthesized and their structures elucidated by IR, 1H-NMR spectroscopy and elemental analysis. The cytotoxicity of the 2-thiohydantoin derivatives to rat embryo fibroblasts (F2408) in vitro was determined, and the effects of these compounds on intracellular free Ca2+, [Ca2+]i, were measured by spectrofluorophotometry. Cytotoxicity was determined by metabolic reduction of a tetrazolium salt to a formazan dye (MTT assay). Compounds 4 and 7 showed cytotoxic activity, with IC50 values in the range of 1-1.2 microM. Introduction of either chlorophenyl, metoxyphenyl, nitrophenyl or benzyl groups at C-3 resulted in concentration-dependent cytotoxic effects. Compounds 1-6 at 1 microM or more significantly increased [Ca2+]i in a dose-dependent manner in the cultured fibroblasts. This action may have been mediated through intracellular calcium stores.


A high-fat diet impairs cardiac high-energy phosphate metabolism and cognitive function in healthy human subjects.
High-fat, low-carbohydrate diets are widely used for weight reduction, but they may also have detrimental effects via increased circulating free fatty acid concentrations. We tested whether raising plasma free fatty acids by using a high-fat, low-carbohydrate diet results in alterations in heart and brain in healthy subjects. Men (n = 16) aged 22 ± 1 y (mean ± SE) were randomly assigned to 5 d of a high-fat, low-carbohydrate diet containing 75 ± 1% of calorie intake through fat consumption or to an isocaloric standard diet providing 23 ± 1% of calorie intake as fat. In a crossover design, subjects undertook the alternate diet after a 2-wk washout period, with results compared after the diet periods. Cardiac (31)P magnetic resonance (MR) spectroscopy and MR imaging, echocardiography, and computerized cognitive tests were used to assess cardiac phosphocreatine (PCr)/ATP, cardiac function, and cognitive function, respectively. Compared with the standard diet, subjects who consumed the high-fat, low-carbohydrate diet had 44% higher plasma free fatty acids (P < 0.05), 9% lower cardiac PCr/ATP (P < 0.01), and no change in cardiac function. Cognitive tests showed impaired attention (P < 0.01), speed (P < 0.001), and mood (P < 0.01) after the high-fat, low-carbohydrate diet. Raising plasma free fatty acids decreased myocardial PCr/ATP and reduced cognition, which suggests that a high-fat diet is detrimental to heart and brain in healthy subjects.


Elastic capsules at liquid-liquid interfaces.
We investigate the deformation of elastic microcapsules adsorbed at liquid-liquid interfaces. An initially spherical elastic capsule at a liquid-liquid interface undergoes circumferential stretching due to the liquid-liquid surface tension and becomes lens- or discus-shaped, depending on its bending rigidity. The resulting elastic capsule deformation is qualitatively similar, but distinct from the deformation of a liquid droplet into a liquid lens at a liquid-liquid interface. We discuss the deformed shapes of droplets and capsules adsorbed at liquid-liquid interfaces for a whole range of different surface elasticities: from droplets (only surface tension) deforming into liquid lenses, droplets with a Hookean membrane (finite stretching modulus, zero bending modulus) deforming into elastic lenses, to microcapsules (finite stretching and bending modulus) deforming into rounded elastic lenses. We calculate capsule shapes at liquid-liquid interfaces numerically using shape equations from nonlinear elastic shell theory. Finally, we present theoretical results for the contact angle (or the capsule height) and the maximal capsule curvature at the three phase contact line. These results can be used to infer information about the elastic moduli from optical measurements. During capsule deformation into a lens-like shape, surface energy of the liquid-liquid interface is converted into elastic energy of the capsule shell giving rise to an overall adsorption energy gain by deformation. Soft hollow capsules exhibit a pronounced increase of the adsorption energy as compared to filled soft particles and, thus, are attractive candidates as foam and emulsion stabilizers.


ASSESSMENT OF DISEASE RISK ASSOCIATED WITH POTENTIAL REMOVAL OF ANTHROPOGENIC BARRIERS TO MOJAVE DESERT TORTOISE (GOPHERUS AGASSIZII) POPULATION CONNECTIVITY.
The Mojave Desert tortoise (Gopherus agassizii), federally listed as threatened, has suffered habitat loss and fragmentation due to human activities. Upper respiratory tract disease (URTD), a documented health threat to desert tortoises, has been detected at the Large-Scale Translocation Study Site (LSTS) in southwestern Nevada, US, a fenced recipient site for translocated animals. Our study aimed to 1) estimate prevalence of URTD and Mycoplasma infection at LSTS and three nearby unfenced sites; 2) assess whether Mycoplasma infection status was associated with developing clinical signs of URTD; and 3) determine whether such an association differed between LSTS and unfenced areas. We sampled 421 tortoises in 2016 to describe the current status of these populations. We evaluated three clinical signs of URTD (nasal discharge, ocular discharge, nasal erosions) and determined individual infection status for Mycoplasma agassizii and Mycoplasma testudineum by quantitative PCR and enzyme-linked immunosorbent assay. In 2016, LSTS had the highest prevalence of M. agassizii (25.0%; 33/132), M. testudineum (3.0%; 4/132), and URTD clinical signs (18.9%; 25/132). Controlling for other factors, clinical sign(s) were positively associated with M. agassizii infection (odds ratio [OR]=7.7, P=0.001), and this effect was similar among study sites (P>0.99). There was no association with M. testudineum status (P=0.360). Of the 196 tortoises in a longitudinal comparison of 2011-14 with 2016, an estimated 3.2% converted from M. agassizii-negative to positive during the study period, and incidence was greater at LSTS (P=0.002). Conversion to positive M. agassizii status was associated with increased incidence of clinical signs in subsequent years (OR=11.1, P=0.018). While M. agassizii and URTD are present outside the LSTS, there is a possibility that incidence of Mycoplasma infection and URTD would increase outside LSTS if these populations were to reconnect. Population-level significance of this risk appears low, and any risk must be evaluated against the potential long-term benefits to population viability through increased connectivity.


How accurately do young adults recall childhood pets? A validation study.
Epidemiologic research shows that pets influence human health, demonstrating both protective and deleterious health risks; therefore, valid definitions of pet exposure would enhance research. The authors determined how well young adults aged 18 years report their early childhood pets. Subjects in an established birth cohort from Detroit, Michigan, born in 1987-1989 (n = 820) were asked a series of questions about pets in the home during their first 6 years of life. Pet recall was compared with annual prospectively collected parental report from 12-18 years prior. Exposure to cats was correctly reported on average 86.3% of the time (95% confidence interval: 85.0, 87.5) and dogs 79.2% (95% confidence interval: 77.7, 80.6) of the time (P < 0.01). Cats and dogs were more likely to be underreported than overreported, from as few as 1.8-fold to as many as 8.3-fold (P < 0.05). Reporting differed by sex of the respondent and current pet ownership. No differences were found in reporting by those who experienced allergy symptoms near dogs or cats. Findings suggest good reliability of young adult pet reporting for ages 0-6 years but that childhood pet exposure may need to be assessed separately depending on the participant's sex and the outcome of interest.


Progressive deficit of retrograde axonal transport is associated with the pathogenesis of di-n-butyl dichlorvos axonopathy.
The induction of central-peripheral distal axonopathy in hens singly dosed with some organophosphorus (OP) compounds, such as di-n-butyl-2,2-dichlorovinyl phosphate (DBDCVP), requires greater than 80% organophosphorylation and subsequent intramolecular rearrangement ("aging") of a protein [neuropathy target esterase (NTE)] in the axon. Suprathreshold biochemical reaction, 24 h after dosing with DBDCVP (0.75-1.00 mg/kg s.c.), is shown to be associated with progressive decrement of retrograde axonal transport in sensory and motor fibers. The maximum transport deficit (about 70% reduction) is reached 7 days after DBDCVP, prior to the appearance of axonal degeneration and the onset of clinical signs of neuropathy (day 10-11). By contrast, phenylmethylsulfonyl fluoride (30 mg/kg s.c.), an agent that prevents the development of OP neuropathy by inhibiting NTE without the "aging" reaction, had no effect on axon transport, nerve fiber integrity, or clinical status and, when administered prior to a neurotoxic dose of DBDCVP (1.00 mg/kg s.c.), prevented DBDCVP effects. Paraoxon (0.2 mg/kg s.c.) neither inhibited NTE nor caused deficits in retrograde transport or neuropathy. Taken in concert, these studies demonstrate that induced deficits in retrograde transport are associated with the pathogenesis of OP-induced nerve-fiber degeneration and the threshold-initiating mechanism thereof.


Genital tract infection with Chlamydia trachomatis fails to induce protective immunity in gamma interferon receptor-deficient mice despite a strong local immunoglobulin A response.
CD4+ T cells have been found to play a critical role in immune protection against Chlamydia trachomatis infection. Since both humoral and cell-mediated antichlamydial immunity have been implicated in host protection, the crucial effector functions provided by the CD4+ T cells may rely on Th1 or Th2 functions or both. In the present study, we evaluated the development of natural immunity following vaginal infection with C. trachomatis serovar D in female gamma interferon receptor-deficient (IFN-gammaR-/-) mice with a disrupted Th1 effector system. We found that in comparison with wild-type mice, the IFN-gammaR-/- mice exhibited a severe ascending primary infection of prolonged duration which stimulated almost 10-fold-stronger specific local immunoglobulin A (IgA) and IgG responses in the genital tract. Following resolution of the primary infection and despite the augmented antibody responses to chlamydiae, the IFN-gammaR-/- mice were completely unprotected against reinfection, suggesting that local antibodies play a subordinate role in host protection against chlamydial infection. Immunohistochemical analysis of frozen sections of the genital tract revealed many CD4+ T cells in the IFN-gammaR-/- mice, with a dominance of interleukin 4-containing cells in mice following resolution of the secondary infection. However, in contrast to the findings with wild-type mice, the typical clusters of CD4+ T cells were not found in the IFN-gammaR-/- mice. Few and similarly distributed CD8+ T cells were observed in IFN-gammaR-/- and wild-type mice. Whereas chlamydia-infected macrophages from wild-type mice had no inclusion bodies (IB) and produced significant amounts of nitric oxide (NO) in the presence of IFN-gamma, macrophages from IFN-gammaR-/- mice contained many IB but no NO. These results indicate that CD4+ Th1 cells and IFN-gamma, rather than local antibodies, are critical elements in host immune protection stimulated by a natural ascending C. trachomatis infection in the female genital tract.


Choice of mate, a reproductive isolating mechanism between Schistosoma intercalatum and S. mansoni in mixed infections.
In Africa, the two human schistosome parasites, Schistosoma intercalatum and S. mansoni, can be found sympatrically, which poses the problem of the nature of their reproductive isolation. In this paper, experimental studies of mating behaviour in mixed infections in mice were carried out. Simultaneous infections without choice of mate showed an absence of any physiological barriers preventing encounter and mating of the two species on the same host. Whenever choice was possible, mating did not occur in a random way, but preferentially between partners of the same species (there was a significant prevalence of homospecific pairs). S. mansoni exhibited greater specific mate recognition than S. intercalatum. Moreover, in the absence of S. mansoni female worms, S. mansoni male worms were as competitive as S. intercalatum male worms in pairing with S. intercalatum female worms. It appears from this study that the choice of mate is a reproductive isolating factor between the two species.


Cardiovascular and gill microcirculatory effects of endothelin-1 in atlantic cod: evidence for pillar cell contraction
Endothelin-1 (ET-1) has been shown to cause a considerable increase in the vascular resistance of fish gills. In trout, recent evidence suggest that this is the result of pillar cell contraction in the gill lamellae. Using epi-illumination microscopy to observe the gill lamellae of anaesthetised Atlantic cod (Gadus morhua), we show that ET-1 (100 ng kg-1, injected into the ventral aorta) causes an increase in pillar cell diameter, consistent with pillar cell contraction, and a shift of intralamellar blood flow from the lamellar sheet to the outer marginal channels. Simultaneously, there was an increase in ventral aortic blood pressure, a reduction in cardiac output, an increase in gill vascular resistance and a reduction in the oxygen partial pressure of venous blood. All these effects were blocked by the ETA/ETB receptor antagonist bosentan (5 mg kg-1). Pillar cell contraction is likely to be a mechanism for matching the functional respiratory surface area with the instantaneous respiratory needs of the fish.


Neoprene Orthopaedic Supports: An Underrecognised Cause of Allergic Contact Dermatitis.
Thioureas, often contained within neoprene to provide water resistance, are an important cause of allergic contact dermatitis (ACD) in those who use neoprene products. We wish to present three cases of thiourea-induced ACD from three different orthopaedic supports containing neoprene. The first case was a 67-year-old woman who developed an itchy rash on her heel three weeks after using a neoprene insole for plantar fasciitis. The second case was a 47-year-old man who developed an itchy rash on his wrist after wearing neoprene wrist splints for psoriatic arthropathy. The third case was a 77-year-old woman who experienced a severe erythematous rash with blistering from a neoprene elbow brace she received following a humeral fracture. All patients were patch tested to the British Society of Cutaneous Allergy Standard and rubber series and a cut piece from all the relevant supports. At 96 hours, all patients had a + reaction to mixed dialkylthiourea, diethylthiourea, and the supports' material. No other positive patch test reactions were identified. As neoprene is fast becoming one of the most popular materials used for orthopaedic supports, awareness of this reaction and close liaison between dermatologists and orthopaedic surgeons are therefore essential to allow for early recognition of this complication.


Psychiatric Disorders in Young Adults Diagnosed with Juvenile Fibromyalgia in Adolescence.
Adolescents with juvenile-onset fibromyalgia (JFM) have increased rates of psychiatric disorders, but to our knowledge no studies have examined psychiatric disorders in adolescents with JFM when they enter young adulthood. This study examined the prevalence of psychiatric disorders in young adults diagnosed with JFM during adolescence and the relationship between mental health diagnoses and physical functioning. Ninety-one young adults (mean age 21.60, SD 1.96) with a history of JFM being followed as part of a prospective longitudinal study and 30 matched healthy controls (mean age 21.57, SD 1.55) completed a structured interview of psychiatric diagnoses and a self-report measure of physical impairment. Young adults with a history of JFM were more likely to have current and lifetime histories of anxiety disorders (70.3% and 76.9%, respectively) compared with controls (33.3% for both, both p < 0.001). Individuals with JFM were also more likely to have current and lifetime histories of major mood disorders (29.7% and 76.9%, respectively) compared with controls (10% and 40%, p < 0.05). The presence of a current major mood disorder was significantly related to impairment in physical functioning [F (1, 89) = 8.30, p < 0.01] and role limitations attributable to a physical condition [F (1, 89) = 7.09, p < 0.01]. Psychiatric disorders are prevalent in young adulthood for individuals with a history of JFM, and a current major mood disorder is associated with greater physical impairment. Greater attention to early identification and treatment of mood disorders in patients with JFM is warranted.


The Prognostic Value of Pre-Treatment Circulating Biomarkers of Systemic Inflammation (CRP, dNLR, YKL-40, and IL-6) in Vulnerable Older Patients with Metastatic Colorectal Cancer Receiving Palliative Chemotherapy-The Randomized NORDIC9-Study.
Appropriate patient selection for palliative chemotherapy is crucial in patients with metastatic colorectal cancer (mCRC). We investigated the prognostic value of C-reactive protein (CRP), derived neutrophil-to-lymphocyte ratio (dNLR), Interleukin (IL)-6, and YKL-40 on progression-free survival (PFS) and overall survival (OS) in the NORDIC9 cohort. The randomized NORDIC9-study included patients ≥70 years with mCRC not candidates for standard full-dose combination chemotherapy. Participants received either full-dose S1 (Teysuno) or a dose-reduced S1 plus oxaliplatin. Blood samples were collected at baseline and biomarkers were dichotomized according to standard cut-offs. Multivariable analyses adjusted for age, sex, ECOG performance status, and treatment allocation; furthermore, C-statistics were estimated. In total, 160 patients with a median age of 78 years (IQR: 76−81) were included between 2015 and 2017. All investigated biomarkers were significantly elevated in patients with either weight loss, ≥3 metastatic sites, or primary tumor in situ. In multivariable analyses, all markers showed significant association with OS; the highest HR was observed for CRP (HR = 3.40, 95%CI: 2.20−5.26, p < 0.001). Regarding PFS, statistically significant differences were found for CRP and IL-6, but not for dNLR and YKL-40. Applying C-statistics, CRP indicated a good prognostic model for OS (AUC = 0.72, 95%CI: 0.67−0.76). CRP is an easily available biomarker, which may support therapeutic decision-making in vulnerable older patients with mCRC.


Compassion-focused imagery reduces shame and is moderated by shame, self-reassurance and multisensory imagery vividness.
Compassion-focused imagery (CFI) is an emotion-regulation technique involving visualization of a person, animal or object offering one compassion, to generate feelings of safeness. It is proven to increase self-compassion and reduce negative affect. This study explores two hypotheses not previously investigated: i) which sensory modalities can stimulate compassionate affect; and ii) whether presentation of pictorial stimuli can enhance CFI. Additionally, we examine iii) whether CFI can reduce shame and iv) whether self-criticism inhibits CFI, since previous studies have involved small samples or methodological limitations. After completing measures of self-criticism, selfreassurance and imagery abilities in five sensory modalities, participants (n=160) were randomly assigned to look at compassionate images during CFI (visual input), compassionate images before CFI (priming), or abstract images (control). Participants trialled CFI then rated compassionate affect and completed open-response questions. Before and after CFI, participants recalled a shame-based memory and rated state shame. Correlational analyses explored whether self-criticism, self-reassurance, and multisensory imagery abilities moderated outcomes. CFI significantly reduced shame regarding a recalled memory, particularly for those high in shame. Compassionate affect was predicted by imagery vividness in visual and bodily-sensation modalities. Self-criticism predicted poorer CFI In multiple regressions, self-reassurance predicted poorer CFI outcomes but self-criticism did not. Between-group effects did not emerge. Qualitative data suggested that pictures helped some participants but hindered others. CFI is a promising technique for shame-prone clients, but may be challenging for those with low imagery abilities or unfamiliar with self-reassurance. Multiple senses should be engaged.


Slowly progressive, angiotensin II-independent glomerulosclerosis in human (pro)renin receptor-transgenic rats.
For defining the pathogenic effects of the (pro)renin receptor-transgenic rat, strains that overexpressed the human receptor were generated. Although transgenic rats were normotensive and euglycemic and had a renal angiotensin II (AngII) level that was comparable to that of wild-type rats, transgenic rats developed proteinuria with aging and significant glomerulosclerosis at 28 wk of age. In kidneys of 28-wk-old transgenic rats, mitogen-activated protein kinases (MAPK) were activated without recognizable tyrosine phosphorylation of the EGF receptor, and expression of TGF-beta1 was enhanced. In vivo infusion of the (pro)renin receptor blocker peptide (formerly handle region decoy peptide) significantly inhibited the development of glomerulosclerosis, proteinuria, MAPK activation, and TGF-beta1 expression in the kidneys, but the angiotensin-converting enzyme inhibitor did not attenuate these changes despite a significant decrease in the renal AngII level. In addition, recombinant rat prorenin stimulated MAPK activation in the human receptor-expressed cultured cells, but human receptor was unable to evoke the enzyme activity of rat prorenin. Thus, human (pro)renin receptor elicits slowly progressive nephropathy by AngII-independent MAPK activation in rats. This study clearly provided in vivo evidence for the AngII-independent MAPK activation by human (pro)renin receptor and induction of glomerulosclerosis with increased TGF-beta1 expression.


The comparison of alternative smoothing methods for fitting non-linear exposure-response relationships with Cox models in a simulation study.
We examined the behavior of alternative smoothing methods for modeling environmental epidemiology data. Model fit can only be examined when the true exposure-response curve is known and so we used simulation studies to examine the performance of penalized splines (P-splines), restricted cubic splines (RCS), natural splines (NS), and fractional polynomials (FP). Survival data were generated under six plausible exposure-response scenarios with a right skewed exposure distribution, typical of environmental exposures. Cox models with each spline or FP were fit to simulated datasets. The best models, e.g. degrees of freedom, were selected using default criteria for each method. The root mean-square error (rMSE) and area difference were computed to assess model fit and bias (difference between the observed and true curves). The test for linearity was a measure of sensitivity and the test of the null was an assessment of statistical power. No one method performed best according to all four measures of performance, however, all methods performed reasonably well. The model fit was best for P-splines for almost all true positive scenarios, although fractional polynomials and RCS were least biased, on average.


Effects on protein and mRNA expression levels of p53 induced by fluoride in human embryonic hepatocytes.
We investigated the effects of protein and mRNA expression levels on p53 induced by fluoride in human embryo hepatocyte L-02 cells. The protein and mRNA levels of p53 in L-02 cells were measured after in vitro cultured L-02 was exposed to sodium fluoride at different doses (40, 80, and 160 microg/ml) for 24 h. The results showed that the cell survival rate of L-02 cells in the high dose fluoride group was significantly lower than that of the control group. The protein expression levels of p53 in the middle and high dose fluoride group were significantly higher than in the control group and elevated with increasing fluoride concentration. The mRNA expression levels of p53 in the fluoride groups were markedly higher than in the control group. The mRNA expression level of p53 in the high dose fluoride group was however lower compared to the middle dose fluoride group, but similar to the low dose fluoride group. These finding suggest that fluoride can decrease the L-02 cells survival rate and induce protein and mRNA expressions of p53; however, there is no consistency between the protein expression level of p53 and the mRNA expression level.


Visualizing blood cell emergence from aortic endothelium.
Three recent Nature papers use time-lapse confocal imaging to visualize the birth of blood cells from the aortic endothelium. Two studies (Bertrand et al., 2010; Kissa and Herbomel, 2010) utilize the zebrafish embryo, while the third (Boisset et al., 2010) develops a novel technique to image the mouse aorta.


Medical therapy vs surgery for recurrent acute rhinosinusitis.
Treatment indications for recurrent acute rhinosinusitis (RARS) remain poorly defined. We studied outcomes of medical vs surgical treatment of RARS, anatomic variants associated with RARS, and factors predicting crossover from medical to surgical treatment. A total of 220 RARS patients treated between 2006 and 2014 were retrospectively divided into 3 cohorts: medical only (MED); surgical only (SURG); or medical crossing over into surgical (CROSS). Twenty-two item Sino-Nasal Outcome Test (SNOT-22) scores, modified Lund-Kennedy endoscopy scores, and prevalence of anatomic variants by computed tomography (CT) were compared. A total of 220 CT scans obtained for non-sinus indications served as controls. A logistic regression model was used for analysis. The mean baseline SNOT-22 scores for all cohorts were similar (MED = 48, SURG = 49, CROSS = 45, p < 0.0001). The SURG cohort showed greater reduction of SNOT-22 scores compared to the MED cohort at 3, 6, and 12 months follow-up (p < 0.0001). The crossover cohort converted to surgery after escalation of SNOT-22 score by a mean of 15 points (p < 0.03), and showed significant reduction postoperatively (p < 0.0001). Haller cell (odds ratio [OR] 3.9; p < 0.0001), concha bullosa (OR 3.7; p < 0.003), and accessory ostium (OR 2.2; p < 0.01) were more common in the entire RARS group vs controls; however, there were no inter-cohort differences in prevalence. RARS patients can benefit from both medical and surgical treatment strategies, but surgical treatment results in greater symptomatic improvement compared to medical treatment. Patients cross over from medical to surgical treatment when SNOT-22 scores escalate by a mean of 15 points. Haller cell, concha bullosa, and accessory ostium are associated with RARS but are equally common in medical, surgical, and crossover cohorts.


Trust Your Good Friends: Source-Free Domain Adaptation by Reciprocal Neighborhood Clustering.
Domain adaptation (DA) aims to alleviate the domain shift between source domain and target domain. Most DA methods require access to the source data, but often that is not possible (e.g., due to data privacy or intellectual property). In this paper, we address the challenging source-free domain adaptation (SFDA) problem, where the source pretrained model is adapted to the target domain in the absence of source data. Our method is based on the observation that target data, which might not align with the source domain classifier, still forms clear clusters. We capture this intrinsic structure by defining local affinity of the target data, and encourage label consistency among data with high local affinity. We observe that higher affinity should be assigned to reciprocal neighbors. To aggregate information with more context, we consider expanded neighborhoods with small affinity values. Furthermore, we consider the density around each target sample, which can alleviate the negative impact of potential outliers. In the experimental results we verify that the inherent structure of the target features is an important source of information for domain adaptation. We demonstrate that this local structure can be efficiently captured by considering the local neighbors, the reciprocal neighbors, and the expanded neighborhood. Finally, we achieve state-of-the-art performance on several 2D image and 3D point cloud recognition datasets.


Extraordinarily widespread and fantastically complex: comparative biology of endosymbiotic bacterial and fungal mutualists of insects.
Endosymbiosis is a pervasive, powerful force in arthropod evolution. In the recent literature, bacterial symbionts of insects have been shown to function as reproductive manipulators, nutritional mutualists and as defenders of their hosts. Fungi, like bacteria, are also frequently associated with insects. Initial estimates suggest that insect-fungal endosymbionts are hyperdiverse, yet there has been comparatively little research investigating the roles that fungi play in their insect hosts. In many systems in which the bacterial symbionts are well-characterized, the possible presence of fungi has been routinely ignored. Why has there been so little research on this important group of symbionts? Here, we explore the differences between fungal and bacterial endosymbiotic insect mutualists. We make predictions about why a bacterium or fungus might be found associated with an insect host given particular ecological, physiological, or evolutionary conditions. We also touch on the various hurdles for studying fungal vs. bacterial endosymbionts and potential future research directions.


On-line orthographic influences on spoken language in a semantic task.
Literacy changes the way the brain processes spoken language. Most psycholinguists believe that orthographic effects on spoken language are either strategic or restricted to meta-phonological tasks. We used event-related brain potentials (ERPs) to investigate the locus and the time course of orthographic effects on spoken word recognition in a semantic task. Participants were asked to decide whether a given word belonged to a semantic category (body parts). On no-go trials, words were presented that were either orthographically consistent or inconsistent. Orthographic inconsistency (i.e., multiple spellings of the same phonology) could occur either in the first or the second syllable. The ERP data showed a clear orthographic consistency effect that preceded lexical access and semantic effects. Moreover, the onset of the orthographic consistency effect was time-locked to the arrival of the inconsistency in a spoken word, which suggests that orthography influences spoken language in a time-dependent manner. The present data join recent evidence from brain imaging showing orthographic activation in spoken language tasks. Our results extend those findings by showing that orthographic activation occurs early and affects spoken word recognition in a semantic task that does not require the explicit processing of orthographic or phonological structure.


Genomic comparison of the ants Camponotus floridanus and Harpegnathos saltator.
The organized societies of ants include short-lived worker castes displaying specialized behavior and morphology and long-lived queens dedicated to reproduction. We sequenced and compared the genomes of two socially divergent ant species: Camponotus floridanus and Harpegnathos saltator. Both genomes contained high amounts of CpG, despite the presence of DNA methylation, which in non-Hymenoptera correlates with CpG depletion. Comparison of gene expression in different castes identified up-regulation of telomerase and sirtuin deacetylases in longer-lived H. saltator reproductives, caste-specific expression of microRNAs and SMYD histone methyltransferases, and differential regulation of genes implicated in neuronal function and chemical communication. Our findings provide clues on the molecular differences between castes in these two ants and establish a new experimental model to study epigenetics in aging and behavior.


An overview on upper respiratory tract infections and bacteriotherapy as innovative therapeutic strategy.
The aim of this review is to describe the most common recurring and chronic upper respiratory tract infections (URTI) in children and discuss the role of bacterial interference and bacteriotherapy in their prevention and treatment. A literature review has been performed on the following topics: acute otitis media, adenoiditis, tonsillitis, rhinosinusitis, microbiotics and the role of bacterial interference, and bacteriotherapy in the prevention and treatment of URTI. Research studies into the characteristics of the microbiological flora and its role in the pathogenesis of URTI have focused on a single pathogen, on resistance to and ineffectiveness of antibiotic therapies, or on the persistence of bacterial biofilm. Recent evidence supports a central role of the existing microbial ecosystem in the pathogenesis of respiratory disease. In light of this, new therapeutic approaches include the implantation and persistence within the normal microflora of relatively innocuous "effector" bacteria that can competitively exclude or prevent the outgrowth of potentially disease-causing bacteria. Recently, a retrospective and observational study demonstrated that S. salivarius 24SMB and S. oralis 89a nasal spray could be effective in the prevention of recurrent otitis media in a real-life setting. Other studies have focused on the role of bacteriotherapy in children with beneficial effects in the prevention of URTI. The results of previous studies on the role of bacteriotherapy in paediatric URTI suggest that the use of bacterial interference phenomena through bacteriotherapy is a feasible, safe approach and deserves proper consideration as a promising therapeutic strategy against URTI.


Choice of transcripts and software has a large effect on variant annotation.
Variant annotation is a crucial step in the analysis of genome sequencing data. Functional annotation results can have a strong influence on the ultimate conclusions of disease studies. Incorrect or incomplete annotations can cause researchers both to overlook potentially disease-relevant DNA variants and to dilute interesting variants in a pool of false positives. Researchers are aware of these issues in general, but the extent of the dependency of final results on the choice of transcripts and software used for annotation has not been quantified in detail. This paper quantifies the extent of differences in annotation of 80 million variants from a whole-genome sequencing study. We compare results using the RefSeq and Ensembl transcript sets as the basis for variant annotation with the software Annovar, and also compare the results from two annotation software packages, Annovar and VEP (Ensembl's Variant Effect Predictor), when using Ensembl transcripts. We found only 44% agreement in annotations for putative loss-of-function variants when using the RefSeq and Ensembl transcript sets as the basis for annotation with Annovar. The rate of matching annotations for loss-of-function and nonsynonymous variants combined was 79% and for all exonic variants it was 83%. When comparing results from Annovar and VEP using Ensembl transcripts, matching annotations were seen for only 65% of loss-of-function variants and 87% of all exonic variants, with splicing variants revealed as the category with the greatest discrepancy. Using these comparisons, we characterised the types of apparent errors made by Annovar and VEP and discuss their impact on the analysis of DNA variants in genome sequencing studies. Variant annotation is not yet a solved problem. Choice of transcript set can have a large effect on the ultimate variant annotations obtained in a whole-genome sequencing study. Choice of annotation software can also have a substantial effect. The annotation step in the analysis of a genome sequencing study must therefore be considered carefully, and a conscious choice made as to which transcript set and software are used for annotation.


A systematic review of intimate partner violence interventions focused on improving social support and/ mental health outcomes of survivors.
Intimate partner violence (IPV) is a key public health issue, with a myriad of physical, sexual and emotional consequences for the survivors of violence. Social support has been found to be an important factor in mitigating and moderating the consequences of IPV and improving health outcomes. This study's objective was to identify and assess network oriented and support mediated IPV interventions, focused on improving mental health outcomes among IPV survivors. A systematic scoping review of the literature was done adhering to PRISMA guidelines. The search covered a period of 1980 to 2017 with no language restrictions across the following databases, Medline, Embase, Web of Science, PROQUEST, and Cochrane. Studies were included if they were primary studies of IPV interventions targeted at survivors focused on improving access to social support, mental health outcomes and access to resources for survivors. 337 articles were subjected to full text screening, of which 27 articles met screening criteria. The review included both quantitative and qualitative articles. As the focus of the review was on social support, we identified interventions that were i) focused on individual IPV survivors and improving their access to resources and coping strategies, and ii) interventions focused on both individual IPV survivors as well as their communities and networks. We categorized social support interventions identified by the review as Survivor focused, advocate/case management interventions (15 studies), survivor focused, advocate/case management interventions with a psychotherapy component (3 studies), community-focused, social support interventions (6 studies), community-focused, social support interventions with a psychotherapy component (3 studies). Most of the studies, resulted in improvements in social support and/or mental health outcomes of survivors, with little evidence of their effect on IPV reduction or increase in healthcare utilization. There is good evidence of the effect of IPV interventions focused on improving access to social support through the use of advocates with strong linkages with community based structures and networks, on better mental health outcomes of survivors, there is a need for more robust/ high quality research to assess in what contexts and for whom, these interventions work better compared to other forms of IPV interventions.


Varied patterns of catch-up in child growth: Evidence from Young Lives.
The development of human capabilities for many disadvantaged children around the world depends on growth recovery ('catch-up growth'). Here we develop a novel framework that allows different types of catch-up growth to be classified and estimated. We distinguish between catch-up in the mean of a group toward that of a healthy reference population versus catch-up within the group. We show these different growth types can be tested in a unified setting using a latent growth framework. We apply the results to four developing countries, using longitudinal data on 7641 children collected over the period 2002-2013. The results show catch-up growth rates are generally modest but vary significantly between countries, and that local environmental factors are material to variation in child growth trajectories. The paper discusses the benefits of the new framework versus current methods, shows that the method is feasible, and suggests they call for intervention designs that are sensitive to community and country contexts.


Glucose metabolism of malignant cells is not regulated by transketolase-like (TKTL)-1.
An isoenzyme of transketolase, transketolase-like (TKTL)-1, has been hypothesized to play a pivotal role in the pathophysiology of malignant tumors. Available data are based on the detection of the putative TKTL-1 protein with one particular mouse monoclonal anti-TKTL-1 antibody, clone JFC12T10. In this study it was demonstrated that a) JFC12T10 detects multiple unspecific bands in Western blots, b) a 75-kDa band hitherto referred to as TKTL-1 corresponds to a nuclear protein and c) immunohistochemical detection of TKTL-1 in benign leiomyomas yields an expression pattern identical to that found in a variety of malignant tumors. In RT-PCR assays, using three different primer pairs for transketolase, TKTL-1 and yet another isogene of transketolase, TKTL-2, a relevant expression of TKTL-1 was not detectable in any of the 6 malignant tumor cell lines investigated (MCF-7, A549, HeLa, HT1080, M21 and TF-1). Expression levels of TKTL-1 were rather similar to those found for TKTL-2, although the latter has never been implicated in malignant disease. On the basis of these data, nutritional recommendations based on a hypothetically TKTL-1 controlled metabolism of tumor cells must be regarded as lacking scientific evidence.


A genome-wide analysis of Arabidopsis Rop-interactive CRIB motif-containing proteins that act as Rop GTPase targets.
The plant-specific Rop family GTPases are versatile molecular switches in many processes during plant growth, development, and responses to the environment. To understand how Rop achieves its functional versatility in signaling, we performed a genome-wide identification of putative Rop targets using a combination of the yeast two-hybrid method, bioinformatic tools, and a robust functional assay in pollen. In this study, we have identified 11 Arabidopsis genes encoding novel proteins, termed RICs (for Rop-interactive CRIB motif-containing proteins), that contain a CRIB (for Cdc42/Rac-interactive binding) motif required for their specific interaction with GTP-bound Rop1. RICs are divergent and classified into five groups that share little sequence homology outside of the conserved Rop-interactive domain. Overexpression in tobacco pollen tubes of the nine Ric genes that are expressed in Arabidopsis pollen causes distinct phenotypes, implying distinct functions for various RICs. RIC3 (group III) and RIC4 (group V) both cause depolarized growth like Rop1 and display Rop1-enhanced localization to the tip of pollen tubes, suggesting that these RICs may be two distinct targets of Rop1. In contrast, RIC10 (group I) promotes pollen tube elongation but does not affect pollen tube growth polarity and shows Rop1-independent localization to the cytoplasm, suggesting that RIC10 may participate in a Rop1-independent pathway probably controlled by a different Rop. Expression of all other RICs causes various degrees of growth inhibition in pollen tubes. Furthermore, these inhibitory RICs also exhibit distinct patterns of localization in pollen tubes. Our results suggest that various RICs have evolved to interact with Rops differentially and to perform distinct functions in pollen tubes. Reverse transcriptase-mediated polymerase chain reaction analysis showed that six of the nine RICs are expressed in various parts of Arabidopsis plants. On the basis of these observations, we propose that RICs function as Rop GTPase targets that control various Rop-dependent signaling pathways in plants.


A molecular theory of germinal center B cell selection and division.
The selection of B cells (BCs) in germinal centers (GCs) is pivotal to the generation of high-affinity antibodies and memory BCs, but it lacks global understanding. Based on the idea of a single Tfh-cell signal that controls BC selection and division, experiments appear contradictory. Here, we use the current knowledge on the molecular pathways of GC BCs to develop a theory of GC BC selection and division based on the dynamics of molecular factors. This theory explains the seemingly contradictory experiments by the separation of signals for BC fate decision from signals controlling the number of BC divisions. Three model variants are proposed and experiments are predicted that allow one to distinguish those. Understanding information processing in molecular BC states is critical for targeted immune interventions, and the proposed theory implies that selection and division can be controlled independently in GC reactions.


LOST to follow-up Information in Trials (LOST-IT): a protocol on the potential impact.
Incomplete ascertainment of outcomes in randomized controlled trials (RCTs) is likely to bias final study results if reasons for unavailability of patient data are associated with the outcome of interest. The primary objective of this study is to assess the potential impact of loss to follow-up on the estimates of treatment effect. The secondary objectives are to describe, for published RCTs, (1) the reporting of loss to follow-up information, (2) the analytic methods used for handling loss to follow-up information, and (3) the extent of reported loss to follow-up. We will conduct a systematic review of reports of RCTs recently published in five top general medical journals. Eligible RCTs will demonstrate statistically significant effect estimates with respect to primary outcomes that are patient-important and expressed as binary data. Teams of 2 reviewers will independently determine eligibility and extract relevant information from each eligible trial using standardized, pre-piloted forms. To assess the potential impact of loss to follow-up on the estimates of treatment effect we will, for varying assumptions about the outcomes of participants lost to follow-up (LTFU), calculate (1) the percentage of RCTs that lose statistical significance and (2) the mean change in effect estimate across RCTs. The different assumptions we will test are the following: (1) none of the LTFU participants had the event; (2) all LTFU participants had the event; (3) all LTFU participants in the treatment group had the event; none of those in the control group had it (worst case scenario); (4) the event incidence among LTFU participants (relative to observed participants) increased, with a higher relative increase in the intervention group; and (5) the event incidence among LTFU participants (relative to observed participants) increased in the intervention group and decreased in the control group. We aim to make our objectives and methods transparent. The results of this study may have important implications for both clinical trialists and users of the medical literature.


Structural characterization and reversal of the natural organophosphate resistance of a D-type esterase, Saccharomyces cerevisiae S-formylglutathione hydrolase.
Saccharomyces cerevisiae expresses a 67.8 kDa homodimeric serine thioesterase, S-formylglutathione hydrolase (SFGH), that is 39.9% identical with human esterase D. Both enzymes possess significant carboxylesterase and S-formylglutathione thioesterase activity but are unusually resistant to organophosphate (OP) inhibitors. We determined the X-ray crystal structure of yeast (y) SFGH to 2.3 A resolution by multiwavelength anomalous dispersion and used the structure to guide site-specific mutagenesis experiments addressing substrate and inhibitor reactivity. Our results demonstrate a steric mechanism of OP resistance mediated by a single indole ring (W197) located in an enzyme "acyl pocket". The W197I substitution enhances ySFGH reactivity with paraoxon by >1000-fold ( k i (W197I) = 16 +/- 2 mM (-1) h (-1)), thereby overcoming natural OP resistance. W197I increases the rate of OP inhibition under pseudo-first-order conditions but does not accelerate OP hydrolysis. The structure of the paraoxon-inhibited W197I variant was determined by molecular replacement (2.2 A); it revealed a stabilized sulfenic acid at Cys60. Wild-type (WT) ySFGH is inhibited by thiol reactive compounds and is sensitive to oxidation; thus, the cysteine sulfenic acid may play a role in the regulation of a "D-type" esterase. The structure of the W197I variant is the first reported cysteine sulfenic acid in a serine esterase. We constructed five Cys60/W197I variants and show that introducing a positive charge near the oxyanion hole, W197I/C60R or W197I/C60K, results in a further enhancement of the rates of phosphorylation with paraoxon ( k i = 42 or 80 mM (-1) h (-1), respectively) but does not affect the dephosphorylation of the enzyme. We also characterized three histidine substitutions near the oxyanion hole, G57H, L58H, and M162H, which significantly decrease esterase activity.


Grey nurse shark (Carcharias taurus) diving tourism: Tourist compliance and shark behaviour at Fish Rock, Australia.
Humans can dive with critically endangered grey nurse sharks (Carcharias taurus) along the east coast of Australia. This study investigated both compliance of tourist divers to a code of conduct and legislation and the behaviour of grey nurse sharks in the presence of divers. A total of 25 data collection dives were conducted from December 2008 to January 2009. Grey nurse shark and diver behaviour were documented using 2-min scan samples and continuous observation. The proportion of time spent observing human-shark interactions was 9.4% of total field time and mean human-shark interaction time was 15.0 min. Results were used to gauge the effectiveness of current management practices for the grey nurse shark dive industry at Fish Rock in New South Wales, Australia. Grey nurse shark dive tourists were compliant to stipulations in the code of conduct and legislation (compliance ranged from 88 to 100%). The research detailed factors that may promote compliance in wildlife tourism operations such as the clarity of the stipulations, locality of the target species and diver perceptions of sharks. Results indicated that grey nurse sharks spent the majority of their time milling (85%) followed by active swimming (15%). Milling behaviour significantly decreased in the presence of more than six divers. Distance between sharks and divers, interaction time and number of sharks were not significantly correlated with grey nurse shark school behaviour. Jaw gaping, rapid withdrawal and stiff or jerky movement were the specific behaviours of grey nurse sharks that occurred most frequently and were associated with distance between divers and sharks and the presence of six or more divers. Revision of the number of divers allowed per interaction with a school of grey nurse sharks and further research on the potential impacts that shark-diving tourism may pose to grey nurse sharks is recommended.


Integration of human factors and ergonomics during medical device design and development: it's all about communication.
Manufacturers of interactive medical devices, such as infusion pumps, need to ensure that devices minimise the risk of unintended harm during use. However, development teams face challenges in incorporating Human Factors. The aim of the research reported here was to better understand the constraints under which medical device design and development take place. We report the results of a qualitative study based on 19 semi-structured interviews with professionals involved in the design, development and deployment of interactive medical devices. A thematic analysis was conducted. Multiple barriers to designing for safety and usability were identified. In particular, we identified barriers to communication both between the development organisation and the intended users and between different teams within the development organisation. We propose the use of mediating representations. Artefacts such as personas and scenarios, known to provide integration across multiple perspectives, are an essential component of designing for safety and usability.


Soil invertebrates disrupt carbon flow through fungal networks.
Annual carbon flux through soil respiration is ten times greater than fossil fuel combustion, but its component parts are poorly understood because they are the product of complex multitrophic interactions between soil organisms. A major component of carbon flux from plants to soil occurs through networks of symbiotic arbuscular mycorrhizal fungi. Here, using 13CO2 pulse labeling, we show that natural densities of the numerically dominant fungal feeding invertebrate Protaphorura armata (order Collembola) reduces 13C enrichment of mycorrhizosphere respiration by 32%. Our findings emphasize the importance of multitrophic interactions in regulating respiration of recent plant photosynthate from soil.


CE: Original Research: Napping on the Night Shift: A Two-Hospital Implementation Project.
: Nurses who work the night shift often experience high levels of sleepiness. Napping has been adopted as an effective countermeasure to sleepiness and fatigue in other safety-sensitive industries, but has not had widespread acceptance in nursing. To assess the barriers to successful implementation of night-shift naps and to describe the nap experiences of night-shift nurses who took naps. In this two-hospital pilot implementation project, napping on the night shift was offered to six nursing units for which the executive nursing leadership had given approval. Unit nurse managers' approval was sought, and where granted, further explanation was given to the unit's staff nurses. A nap experience form, which included the Karolinska Sleepiness Scale, was used to assess pre-nap sleepiness level, nap duration and perceived sleep experience, post-nap sleep inertia, and the perceived helpfulness of the nap. Nurse managers and staff nurses were also interviewed at the end of the three-month study period. Successful implementation occurred on only one of the six units, with partial success seen on a second unit. Barriers primarily occurred at the point of seeking the unit nurse managers' approval. On the successful unit, 153 30-minutes naps were taken during the study period. A high level of sleepiness was present at the beginning of 44% of the naps. For more than half the naps, nurses reported achieving either light (43%) or deep (14%) sleep. Sleep inertia was rare. The average score of helpfulness of napping was high (7.3 on a 1-to-10 scale). Nurses who napped reported being less drowsy while driving home after their shift. These data suggest that when barriers to napping are overcome, napping on the night shift is feasible and can reduce nurses' workplace sleepiness and drowsy driving on the way home. Addressing nurse managers' perceptions of and concerns about napping may be essential to successful implementation.


Spline based iterative phase retrieval algorithm for X-ray differential phase contrast radiography.
Differential phase contrast imaging using grating interferometer is a promising alternative to conventional X-ray radiographic methods. It provides the absorption, differential phase and scattering information of the underlying sample simultaneously. Phase retrieval from the differential phase signal is an essential problem for quantitative analysis in medical imaging. In this paper, we formalize the phase retrieval as a regularized inverse problem, and propose a novel discretization scheme for the derivative operator based on B-spline calculus. The inverse problem is then solved by a constrained regularized weighted-norm algorithm (CRWN) which adopts the properties of B-spline and ensures a fast implementation. The method is evaluated with a tomographic dataset and differential phase contrast mammography data. We demonstrate that the proposed method is able to produce phase image with enhanced and higher soft tissue contrast compared to conventional absorption-based approach, which can potentially provide useful information to mammographic investigations.