Effect of High-Heeled Shoes on Balance and Lower-Extremity Biomechanics During Walking in Experienced and Novice High-Heeled Shoe Wearers.
Wearing high-heeled shoes and carrying asymmetrical loads are common in ladies. However, knowledge of the effects of wearing high-heeled shoes on balance and lower-extremity biomechanics in experienced and novice high-heeled shoe wearers is lacking. The study aims to examine the effects of high-heeled shoes and asymmetrical load carrying on joint kinematics and kinetics of the lower extremity during walking as well as balance in experienced and novice high-heeled shoe wearers. Fifteen experienced and 15 novice high-heeled shoes wearers participated in this study. Using a motion analysis system, kinematic and kinetic data were collected while participants walked at their preferred speed in six conditions created from two types of shoes (9-cm high-heeled shoes and flat-heeled shoes) and three weights of symmetrical load (0%, 5%, and 10% of body weight). Stride time and length, step length, double support time, peak joint angles, and joint moments in a sagittal plane were analyzed. Single-leg and tandem-leg stance tests were performed in each condition. Compared with experienced high-heeled shoe wearers, novice high-heeled shoe wearers had longer double support time and shorter stride length during 10% of body weight asymmetrical load walking; walked with greater knee flexion angle, smaller knee range of motion, and smaller ankle dorsiflexor moment; and scored lower in the single-leg and tandem-leg stance tests. Novice high-heeled shoe wearers need to alter their lower-limb joint angles and moments to adjust to high-heeled shoes to achieve balance during gait while carrying an asymmetrical load.


A multiplex assay for the quantification of antibody responses in Staphylococcus aureus infections in mice.
Staphylococcus aureus causes a variety of infections. Knowledge about the physiological role of most S. aureus antigens in colonization and infection is only limited. This can be studied by measuring antigen-specific antibody responses. In this study, we optimized the multiplex microsphere bead-based flow cytometry technique for mouse serum samples. We analysed immunoglobulin G (IgG) levels directed against 26 S. aureus proteins in a single small-volume mouse serum sample. We assessed possible cross reactivity. Furthermore, we analysed serum samples from mice with different types of S. aureus infections caused by different S. aureus strains. The results show that cross reactivity between proteins on microspheres and serum antibodies towards other proteins was limited. We found that lung-infected mice had a higher and broader IgG response than skin-infected mice. Clearly, the site of infection influences the IgG profile. Next, we compared sera from mice with intravenously-induced bacteraemia caused by different S. aureus strains. We showed different IgG responses depending on the causing S. aureus strain. It is concluded that the bead-based multiplex S. aureus antibody assay can be successfully applied to determine the immunogenicity of different S. aureus proteins in relation to the site of infection and the S. aureus strain causing the infection.


Targeting Translational Successes through CANSORT-SCI: Using Pet Dogs To Identify Effective Treatments for Spinal Cord Injury.
Translation of therapeutic interventions for spinal cord injury (SCI) from laboratory to clinic has been historically challenging, highlighting the need for robust models of injury that more closely mirror the human condition. The high prevalence of acute, naturally occurring SCI in pet dogs provides a unique opportunity to evaluate expeditiously promising interventions in a population of animals that receive diagnoses and treatment clinically in a manner similar to persons with SCI, while adhering to National Institutes of Health guidelines for scientific rigor and transparent reporting. In addition, pet dogs with chronic paralysis are often maintained long-term by their owners, offering a similarly unique population for study of chronic SCI. Despite this, only a small number of studies have used the clinical dog model of SCI. The Canine Spinal Cord Injury Consortium (CANSORT-SCI) was recently established by a group of veterinarians and basic science researchers to promote the value of the canine clinical model of SCI. The CANSORT-SCI group held an inaugural meeting November 20 and 21, 2015 to evaluate opportunities and challenges to the use of pet dogs in SCI research. Key challenges identified included lack of familiarity with the model among nonveterinary scientists and questions about how and where in the translational process the canine clinical model would be most valuable. In light of these, we review the natural history, outcome, and available assessment tools associated with canine clinical SCI with emphasis on their relevance to human SCI and the translational process.


Cardiorespiratory fitness and physical activity in people who have rheumatoid arthritis at an increased risk of cardiovascular disease: a cross-sectional study.
Lower cardiorespiratory fitness (CRF) and physical activity (PA) associate with higher cardiovascular disease (CVD) risk, but the relationship between CRF and PA in people who have rheumatoid arthritis (RA) at an increased CVD risk (CVD-RA) is not known. The objectives of this study were to determine the levels of CRF and PA in people who have CVD-RA and to investigate the association of CRF with PA in people who have CVD-RA. A total of 24 consecutive patients (19 women) with CVD-RA (> 4% for 10-year risk of fatal CVD development as calculated using the Systematic Coronary Risk Evaluation)-were included in the study. CRF was assessed with a graded maximal exercise test determining maximal oxygen uptake (VO2max). PA was assessed with an accelerometer to determine the amount of step count, sedentary, light and moderate-to-vigorous physical activity (MVPA) minutes per day. Mean age of patients was 65.3 ± 8.3 years. CRF mean values were 16.3 ± 1.2 ml·kg-1 min-1, mean step count per day was 6033 ± 2256, and the mean MVPA time was 16.7 min per day. Significant positive associations were found for CRF with step count (B = 0.001, P = 0.01) and MVPA time (B = 0.15, P = 0.02); a negative association was found for CRF with sedentary time (B = - 0.02, P = 0.03). CRF is low and is associated with step count, sedentary time and MVPA time in people who have RA at an increased CVD risk.


Perception of suprasegmental features of speech by children with cochlear implants and children with hearing AIDS.
This study assessed perception of suprasegmental features of speech by 30 prelingual children with sensorineural hearing loss. Ten children had cochlear implants (CIs), and 20 children wore hearing aids (HA): 10 with severe hearing loss and 10 with profound hearing loss. Perception of intonation, syllable stress, word emphasis, and word pattern was assessed. Results revealed that the two HA groups significantly outperformed the CI group in perceiving both intonation and stress. Within each group, word pattern was perceived best, and then intonation and emphasis, with syllable stress perceived poorest. No significant correlation emerged between age at implantation and perception of the various suprasegmental features, possibly due to participants' relatively late age at implantation. Results indicated that CI use did not show an advantage over HA use in the perception of suprasegmental features of speech. Future research should continue to explore variables that might improve this perception.


Selective and flexible depletion of problematic sequences from RNA-seq libraries at the cDNA stage.
A major hurdle to transcriptome profiling by deep-sequencing technologies is that abundant transcripts, such as rRNAs, can overwhelm the libraries, severely reducing transcriptome-wide coverage. Methods for depletion of such unwanted sequences typically require treatment of RNA samples prior to library preparation, are costly and not suited to unusual species and applications. Here we describe Probe-Directed Degradation (PDD), an approach that employs hybridisation to DNA oligonucleotides at the single-stranded cDNA library stage and digestion with Duplex-Specific Nuclease (DSN). Targeting Saccharomyces cerevisiae rRNA sequences in Illumina HiSeq libraries generated by the split adapter method we show that PDD results in efficient removal of rRNA. The probes generate extended zones of depletion as a function of library insert size and the requirements for DSN cleavage. Using intact total RNA as starting material, probes can be spaced at the minimum anticipated library size minus 20 nucleotides to achieve continuous depletion. No off-target bias is detectable when comparing PDD-treated with untreated libraries. We further provide a bioinformatics tool to design suitable PDD probe sets. We find that PDD is a rapid procedure that results in effective and specific depletion of unwanted sequences from deep-sequencing libraries. Because PDD acts at the cDNA stage, handling of fragile RNA samples can be minimised and it should further be feasible to remediate existing libraries. Importantly, PDD preserves the original RNA fragment boundaries as is required for nucleotide-resolution footprinting or base-cleavage studies. Finally, as PDD utilises unmodified DNA oligonucleotides it can provide a low-cost option for large-scale projects, or be flexibly customised to suit different depletion targets, sample types and organisms.


Survival of Sporangia of New Clonal Lineages of Phytophthora infestans in Soil Under Semiarid Conditions.
Currently, there is no information on the viability of sporangia in soil of the new metalaxyl-resistant genotypes of Phytophthora infestans in the semiarid Columbia Basin of Washington and potato-growing regions throughout the world. Sporangia of metalaxyl-resistant US-8 and US-11 clonal lineages of P. infestans survived a maximum of 23 to 30 days in a Shano silt loam and a Quincy loamy fine sand. There were no significant differences between soil types in area under the spore survival curve (AUSSC) in two trials, however, sporangia of P. infestans in the Quincy sand had a significantly greater mean maximum days of sporangia survival (MDSS) than did the Shano silt loam in one of two trials. AUSSC and MDSS were significantly greater (P < 0.05) for sporangia in wet soil than in dry soil under shaded conditions. Mean AUSSC and MDSS significantly decreased (P < 0.01) under nonshaded conditions versus shaded conditions. Three metalaxyl-resistant isolates (two US-8 and one US-11) of P. infestans did not significantly differ (P < 0.05) in AUSSC and MDSS.


Energy Efficiency and Health Efficiency of Old and New EU Member States.
Environmental protection and health issues have always been of great concern. This study employed modified Meta-Frontier Dynamic Network Data Envelopment Analysis to explore the environmental pollution effects from energy consumption on the mortality of children and adults, tuberculosis rate, survival rate, and health expenditure efficiencies in 15 old EU states and 13 new EU states from 2010 to 2014. We calculated the overall efficiency scores and technology gap ratios for each old EU and new EU states as well as the efficiencies of non-renewable energy, renewable energy, PM2.5, CO2, labor, GDP, tuberculosis, child mortality, adult mortality, health expenditure efficiency, and survival efficiency at the health stage. The average annual overall efficiencies of the old EU states are higher than that of the new EU states. Whether in terms of energy efficiencies or health efficiencies, the inputs and outputs of the old EU states are always higher than that of the new EU states. Overall, developing countries in Eastern Europe are lagging behind in terms of energy and health efficiencies. At the same time, the efficiency of child mortality is lower than that of adult mortality, and the efficiency of PM2.5 is higher than that of CO2 in both old and new EU states.


Dynamics of the Eukaryotic Replicative Helicase at Lagging-Strand Protein Barriers Support the Steric Exclusion Model.
Progression of DNA replication depends on the ability of the replisome complex to overcome nucleoprotein barriers. During eukaryotic replication, the CMG helicase translocates along the leading-strand template and unwinds the DNA double helix. While proteins bound to the leading-strand template efficiently block the helicase, the impact of lagging-strand protein obstacles on helicase translocation and replisome progression remains controversial. Here, we show that CMG and replisome progressions are impaired when proteins crosslinked to the lagging-strand template enhance the stability of duplex DNA. In contrast, proteins that exclusively interact with the lagging-strand template influence neither the translocation of isolated CMG nor replisome progression in Xenopus egg extracts. Our data imply that CMG completely excludes the lagging-strand template from the helicase central channel while unwinding DNA at the replication fork, which clarifies how two CMG helicases could freely cross one another during replication initiation and termination.


Multicenter evaluation of a new disk agar diffusion method for susceptibility testing of filamentous fungi with voriconazole, posaconazole, itraconazole, amphotericin B, and caspofungin.
The purpose of this study was to correlate inhibition zone diameters, in millimeters (agar diffusion disk method), with the broth dilution MICs or minimum effective concentrations (MECs) (CLSI M38-A method) of five antifungal agents to identify optimal testing guidelines for disk mold testing. The following disk diffusion testing parameters were evaluated for 555 isolates of the molds Absidia corymbifera, Aspergillus sp. (five species), Alternaria sp., Bipolaris spicifera, Fusarium sp. (three species), Mucor sp. (two species), Paecilomyces lilacinus, Rhizopus sp. (two species), and Scedosporium sp. (two species): (i) two media (supplemented Mueller-Hinton agar [2% dextrose and 0.5 microg/ml methylene blue] and plain Mueller-Hinton [MH] agar), (ii) three incubation times (16 to 24, 48, and 72 h), and (iii) seven disks (amphotericin B and itraconazole 10-microg disks, voriconazole 1- and 10-microg disks, two sources of caspofungin 5-microg disks [BBL and Oxoid], and posaconazole 5-microg disks). MH agar supported better growth of all of the species tested (24 to 48 h). The reproducibility of zone diameters and their correlation with either MICs or MECs (caspofungin) were superior on MH agar (91 to 100% versus 82 to 100%; R, 0.71 to 0.93 versus 0.53 to 0.96 for four of the five agents). Based on these results, the optimal testing conditions for mold disk diffusion testing were (i) plain MH agar; (ii) incubation times of 16 to 24 h (zygomycetes), 24 h (Aspergillus fumigatus, A. flavus, and A. niger), and 48 h (other species); and (iii) the posaconazole 5-microg disk, voriconazole 1-microg disk, itraconazole 10-microg disk (for all except zygomycetes), BBL caspofungin 5-microg disk, and amphotericin B 10-microg (zygomycetes only).


Measurement Invariance and Sex and Age Differences of the Big Five Inventory-2: Evidence From the Russian Version.
The Big Five Inventory-2 (BFI-2) is a recently published 60-item questionnaire that measures personality traits within the five-factor model framework. An important aspect of the BFI-2 is that it measures the traits at both the domain and facet levels and also controls acquiescence bias via the balanced number of true- and false-keyed items across the domains and facets. The current research evaluates factorial measurement invariance of a Russian version of the BFI-2 across sex and age within samples of 1,024 university students (Study 1) and 1,029 Internet users (Study 2). Across these samples, men scored lower on the domains of negative emotionality and agreeableness and slightly higher on extraversion. Sex differences were also obtained on various facets. In the Internet sample, age correlated modestly with several Big Five domains in accordance with the well-documented maturity principle. The newly developed Russian version of BFI-2 showed good reliability and validity across both samples. Moreover, random intercept exploratory factor analyses showed that the BFI-2 displayed a hierarchical five-domain-15-facet structure that demonstrated strict measurement invariance across sex and age.


Subject Specific Optimisation of the Stiffness of Footwear Material for Maximum Plantar Pressure Reduction.
Current selection of cushioning materials for therapeutic footwear and orthoses is based on empirical and anecdotal evidence. The aim of this investigation is to assess the biomechanical properties of carefully selected cushioning materials and to establish the basis for patient-specific material optimisation. For this purpose, bespoke cushioning materials with qualitatively similar mechanical behaviour but different stiffness were produced. Healthy volunteers were asked to stand and walk on materials with varying stiffness and their capacity for pressure reduction was assessed. Mechanical testing using a surrogate heel model was employed to investigate the effect of loading on optimum stiffness. Results indicated that optimising the stiffness of cushioning materials improved pressure reduction during standing and walking by at least 16 and 19% respectively. Moreover, the optimum stiffness was strongly correlated to body mass (BM) and body mass index (BMI), with stiffer materials needed in the case of people with higher BM or BMI. Mechanical testing confirmed that optimum stiffness increases with the magnitude of compressive loading. For the first time, this study provides quantitative data to support the importance of stiffness optimisation in cushioning materials and sets the basis for methods to inform optimum material selection in the clinic.


Evaluation of an intraosseous infusion device for the resuscitation of hypovolemic shock.
An intraosseous infusion device designed for the prehospital administration of hypertonic saline-dextran solutions was evaluated by resuscitating hemorrhaged conscious sheep. Eight animals underwent 2 hours of hemorrhagic hypotension (50 mm Hg, bled volume = 43 +/- 7 ml/kg). This was followed by the intraosseous infusion of 200 ml (4-5 ml/kg) of 7.5% NaCl-6% dextran 70 into the bone marrow of the sternum. Results were compared to seven control animals (bled volume = 31 +/- 6 ml/kg) resuscitated through a central venous catheter. Despite the small volumes infused, mean arterial blood pressure and cardiac output were rapidly normalized in both groups by 10 minutes post resuscitation (p less than 0.01). Plasma sodium concentration increased an average of 12 mEq/L and plasma volume was rapidly expanded regardless of route. The metabolic acidosis of hemorrhagic shock was rapidly corrected, pulmonary pressures remained normal, and hypoxemia did not occur after intraosseous resuscitation. The device provided safe and rapid vascular access via the sternal bone marrow space. The use of intraosseous infusion of hypertonic saline dextran solutions via the sternal bone marrow may allow prehospital rescuers to consistently incorporate fluid replacement therapy into 'scoop and run' policies by avoiding the time delays associated with failures in IV access.


Total least-squares reconstruction with wavelets for optical tomography.
In a previous paper [Zhu et al., J. Opt. Soc. Am. A 14, 799 (1997)] an iterative algorithm for obtaining the total least-squares (TLS) solution of a linear system based on the Rayleigh quotient formulation was presented. Here we derive what to our knowledge are the first statistical properties of this solution. It is shown that the Rayleigh-quotient-form TLS (RQF-TLS) estimator is equivalent to the maximum-likelihood estimator when noise terms in both data and operator elements are independent and identically distributed Gaussian. A perturbation analysis of the RQF-TLS solution is derived, and from it the mean square error of the RQF-TLS solution is obtained in closed form, which is valid at small noise levels. We then present a wavelet-based multiresolution scheme for obtaining the TLS solution. This method was employed with a multigrid algorithm to solve the linear perturbation equation encountered in optical tomography. Results from numerical simulations show that this method requires substantially less computation than the previously reported one-grid TLS algorithm. The method also allows one to identify regions of interest quickly from a coarse-level reconstruction and restrict the reconstruction in the following fine resolutions to those regions. Finally, the method is less sensitive to noise than the one-grid TLS and multigrid least-squares algorithms.


Diagnostic accuracy of direct agglutination test, rK39 ELISA and six rapid diagnostic tests among visceral leishmaniasis patients with and without HIV coinfection in Ethiopia.
Diagnosis of a first-time visceral leishmaniasis (VL) infection in Ethiopia is established by use of a rapid diagnostic test (RDT) detecting antibodies against rK39, direct agglutination test (DAT) and microscopy according to the national algorithm. The performance of individual tests and algorithm is variable and depends on several factors, one being HIV status. Limited data are available on the performance of tests in VL-HIV coinfected patients. Assessment of the performance of DAT (ITM-A), rK39 ELISA (Serion) and six RDT (Onsite Leishmania Ab CTK, Antigen ICT Xinjier, IT Leish Biorad, Kalazar Detect Inbios, rK39 IgG1 Coris, rk28 IgG1 Coris) for the diagnosis of VL was done on a panel of 91 stored serum and plasma samples of 'first-episode' suspected VL patients, with HIV coinfection (n = 51) and without (n = 40). A combined reference standard was used: either positive microscopy on tissue aspirates, or in case of negative microscopy, positive PCR results on the aspirate slide. Additionally, endemic healthy controls (n = 20), non-endemic controls (n = 10) and patients with confirmed malaria infection (n = 10) were tested for specificity evaluation. Sensitivities ranged from 69.2% for DAT (applied cut-off ≥ 1/3200) to 92.2% for the Onsite RDT, whereas specificities ranged from 20.0% for Kalazar Antigen ICT to 100% for IT Leish and rK39 IgG1. Sensitivities from all assays decreased upon stratification according to HIV status but was only significantly different for rK39 Serion ELISA (p-value 0.0084) and the Onsite RDT (p-value 0.0159). In conclusion, performance of commercially available assays for VL on samples from Northern-Ethiopian patients varied widely with a substantial decrease in sensitivity in the VL-HIV coinfected group. Clear guidelines on minimal performance criteria of individual tests and algorithms are needed, as well as which reference standard should be used to determine the performance.


Reliability and Variability of tDCS Induced Changes in the Lower Limb Motor Cortex.
Transcranial direct current stimulation (tDCS) is emerging as a promising adjuvant to enhance motor function. However, there has been increasing reservations about the reliability and variability of the neuromodulatory effects evoked by tDCS. The main purpose of this study was to explore the test-retest reliability and inter-individual variability of tDCS of the lower limb M1 and the relationship between transcranial magnetic stimulation (TMS)-related measures and tDCS-induced changes. Fifteen healthy participants received anodal tDCS of the lower limb M1 either when performing a lower limb motor task or when the limb was at rest. Each condition was tested twice. tDCS induced changes in corticomotor excitability of the tibialis anterior muscle were measured using TMS. A repeated measures ANOVA was performed to examine efficacy of tDCS between the two task conditions. Intraclass correlation coefficients (ICC) and variance component analyses were performed to examine reliability and variability respectively. A significant increase in in corticomotor excitability was noted for the tDCS-task condition at 140% active motor threshold (AMT) and when comparing recruitment curve slopes, but not at 120% and 130% AMT. Overall, ICC values between testing days for each stimulation condition ranged from 0.6-0.9. Higher ICCs were seen for higher TMS intensities (140% AMT) and recruitment curve slopes. Inter-individual variability contributed to 34% of the exhibited variance. Our data suggest that the TMS-related measure used to assess neuromodulation after tDCS has an effect on its perceived test-retest reliability and inter-individual variability. Importantly, we noticed that a high reliability and low variability does not necessarily indicate clinical efficacy of tDCS as some participants showed little to no modulation of corticomotor excitability consistently.


The structure and specificity of Escherichia coli maltose acetyltransferase give new insight into the LacA family of acyltransferases.
The crystallographic three-dimensional structure of the Escherichia coli maa gene product, previously identified as a maltose O-acetyltransferase (MAT) [Brand, B., and Boos, W. (1991) J. Biol. Chem. 266, 14113-14118] has been determined to 2.15 A resolution by the single anomalous dispersion method using data from a crystal cocrystallized with trimethyllead acetate. It is shown here that MAT acetylates glucose exclusively at the C6 position and maltose at the C6 position of the nonreducing end glucosyl moiety. Furthermore, MAT shows higher affinity toward artificial substrates containing an alkyl or hydrophobic chain as well as a glucosyl unit. The presence of a long hydrophobic patch near the acceptor site provides the structural explanation for this preference. The three-dimensional structure reveals the expected trimeric left-handed parallel beta-helix structure found in all other known hexapeptide repeat enzymes. In particular, the structure shows similarities both overall and at the putative active site to the recently determined structure of galactoside acetyltransferase (GAT), the lacA gene product [Wang, X.-G., Olsen, L. R., and Roderick, S. L. (2002) Structure 10, 581-588]. The structure, together with the new biochemical data, suggests that GAT and MAT are more closely related than previously thought and might have similar cellular functions. However, while GAT is specific for acetylation of galactosyl units, MAT is specific for glucosyl units and is able to acetylate maltooligosaccharides, an important property for biotechnological applications. Structural differences at the acceptor site reflect the differences in substrate specificity.


Does EMDR Therapy Have an Effect on Memories of Emotional Abuse, Neglect and Other Types of Adverse Events in Patients with a Personality Disorder? Preliminary Data.
Little is known about the effectiveness of trauma-focused therapies for memories of events not meeting the A-criterion of post-traumatic stress disorder (PTSD). Determining the effect of EMDR therapy on memories of emotional abuse, neglect and other types of adverse events in patients with a personality disorder (PD). We conducted a secondary analysis of the data from our study, which aimed to determine the effectiveness of five sessions of EMDR therapy in 49 patients with a PD. Patients were divided into three different groups depending on their most prevalent type of adverse event. Data were analyzed with Generalized Estimating Equations. Of all patients, 49% reported emotional neglect, 22.4% emotional abuse and 26.5% other types. Only one patient reported memories that predominantly fulfilled the A-criterion of PTSD. After five sessions of EMDR therapy, medium to large treatment effects for memories related to neglect (ds between 0.52 and 0.79), medium treatment effects for memories involving emotional abuse (ds between 0.18 and 0.59) and other types of adverse events were found (ds between 0.18 and 0.53). No significant differences in symptom reduction associated with the application of EMDR therapy among memories involving these three different types of adverse events could be revealed. The results support the notion that EMDR therapy is not only an effective therapy for memories related to A-criteria-worthy events, but that it also has a symptom-reducing effect on memories involving other types of adverse events. This suggests that EMDR might be a valuable addition to the treatment of PD without PTSD.


Aspalathin Protects the Heart against Hyperglycemia-Induced Oxidative Damage by Up-Regulating Nrf2 Expression.
Aspalathin (ASP) can protect H9c2 cardiomyocytes against high glucose (HG)-induced shifts in myocardial substrate preference, oxidative stress, and apoptosis. The protective mechanism of ASP remains unknown. However, as one of possible, it is well known that phytochemical flavonoids reduce oxidative stress via nuclear factor (erythroid-derived 2)-like 2 (Nrf2) activation resulting in up-regulation of antioxidant genes and enzymes. Therefore, we hypothesized that ASP protects the myocardium against HG- and hyperglycemia-induced oxidative damage by up-regulating Nrf2 expression in H9c2 cardiomyocytes and diabetic (db/db) mice, respectively. Using an oxidative stress RT² Profiler PCR array, ASP at a dose of 1 µM was demonstrated to protect H9c2 cardiomyocytes against HG-induced oxidative stress, but silencing of Nrf2 abolished this protective response of ASP and exacerbated cardiomyocyte apoptosis. Db/db mice and their non-diabetic (db/+) littermate controls were subsequently treated daily for six weeks with either a low (13 mg/kg) or high (130 mg/kg) ASP dose. Compared to nondiabetic mice the db/db mice presented increased cardiac remodeling and enlarged left ventricular wall that occurred concomitant to enhanced oxidative stress. Daily treatment of mice with ASP at a dose of 130 mg/kg for six weeks was more effective at reversing complications than both a low dose ASP or metformin, eliciting enhanced expression of Nrf2 and its downstream antioxidant genes. These results indicate that ASP maintains cellular homeostasis and protects the myocardium against hyperglycemia-induced oxidative stress through activation of Nrf2 and its downstream target genes.


Phacoemulsification of the rock-hard dense nuclear cataract: Options and recommendations.
We describe the essential steps in the successful phacoemulsification of the rock-hard, dense cataract. Appropriate and directed preoperative history, physical examination, and diagnostics allow the surgeon to select the best incision, anesthesia, and intended surgical technique for a given dense nuclear challenge. Hard nucleus-specific approaches for hydrodissection, pupil management, and zonular protection then allow the surgeon to approach the rock-hard nucleus with maximum safety. Dense nuclear dismantling options are then discussed in detail along with fluidic and power modulation considerations. Various specific phacoemusification machine settings for rock-hard cataracts from the authors representing several different phaco systems are then presented. The combination of these steps and considerations allow a more successful dense cataract removal and potential restoration of vision for patients. This paper represents the collective experience and advice of the Challenging and Complex Cataract Surgery Subcommittee.


Effects of Acupuncture on Anxiety Levels and Prefrontal Cortex Activity Measured by Near-Infrared Spectroscopy: A Pilot Study.
There is increasing evidence that acupuncture is useful in treating somatic and psychological disorders caused by stress; however, the physiological basis of the effect remains unclear. In the present study, we evaluated the effect of acupuncture on psychological conditions (i.e., anxiety) and prefrontal cortex (PFC) activity. We studied 10 patients with anxiety disorders and measured anxiety levels by means of the State-Trait Anxiety Inventory (STAI), including state anxiety (STAI-1) and trait anxiety (STAI-2). Employing a two-channel NIRS device, we measured oxy-Hb concentration in the bilateral PFC at rest, and evaluated asymmetry of the PFC activity by calculating the Laterality Index at Rest (LIR). The patients were treated by acupuncture at Yui Clinic in Osaka. The treatment significantly decreased the STAI-1 score (p<0.001), but not the STAI-2 score (p>0.05). The NIRS measurements indicated the presence of spontaneous oscillations of oxy-Hb in the bilateral PFC at rest before and after the treatment. Notably LIR decreased significantly in 7 out of the 10 subjects (p<0.01), while 3 subjects showed an increasing tendency. The present pilot study indicates that acupuncture is effective in decreasing anxiety levels in patients with anxiety disorders. Our NIRS data suggest that acupuncture may alter the balance of PFC activity at rest, resulting in relaxation effects. Our NIRS data suggest that acupuncture changes the balance of PFC activity toward left-dominant, resulting in relaxation effects on the patients.


Permitted and forbidden sets in symmetric threshold-linear networks.
The richness and complexity of recurrent cortical circuits is an inexhaustible source of inspiration for thinking about high-level biological computation. In past theoretical studies, constraints on the synaptic connection patterns of threshold-linear networks were found that guaranteed bounded network dynamics, convergence to attractive fixed points, and multistability, all fundamental aspects of cortical information processing. However, these conditions were only sufficient, and it remained unclear which were the minimal (necessary) conditions for convergence and multistability. We show that symmetric threshold-linear networks converge to a set of attractive fixed points if and only if the network matrix is copositive. Furthermore, the set of attractive fixed points is nonconnected (the network is multiattractive) if and only if the network matrix is not positive semidefinite. There are permitted sets of neurons that can be coactive at a stable steady state and forbidden sets that cannot. Permitted sets are clustered in the sense that subsets of permitted sets are permitted and supersets of forbidden sets are forbidden. By viewing permitted sets as memories stored in the synaptic connections, we provide a formulation of long-term memory that is more general than the traditional perspective of fixed-point attractor networks. There is a close correspondence between threshold-linear networks and networks defined by the generalized Lotka-Volterra equations.


Can Bayesian Theories of Autism Spectrum Disorder Help Improve Clinical Practice?
Diagnosis and individualized treatment of autism spectrum disorder (ASD) represent major problems for contemporary psychiatry. Tackling these problems requires guidance by a pathophysiological theory. In this paper, we consider recent theories that re-conceptualize ASD from a "Bayesian brain" perspective, which posit that the core abnormality of ASD resides in perceptual aberrations due to a disbalance in the precision of prediction errors (sensory noise) relative to the precision of predictions (prior beliefs). This results in percepts that are dominated by sensory inputs and less guided by top-down regularization and shifts the perceptual focus to detailed aspects of the environment with difficulties in extracting meaning. While these Bayesian theories have inspired ongoing empirical studies, their clinical implications have not yet been carved out. Here, we consider how this Bayesian perspective on disease mechanisms in ASD might contribute to improving clinical care for affected individuals. Specifically, we describe a computational strategy, based on generative (e.g., hierarchical Bayesian) models of behavioral and functional neuroimaging data, for establishing diagnostic tests. These tests could provide estimates of specific cognitive processes underlying ASD and delineate pathophysiological mechanisms with concrete treatment targets. Written with a clinical audience in mind, this article outlines how the development of computational diagnostics applicable to behavioral and functional neuroimaging data in routine clinical practice could not only fundamentally alter our concept of ASD but eventually also transform the clinical management of this disorder.


SU-E-T-242: Effect of Skull Surgical Clips and Bone Artifacts on Treatment Planning for Brain Tumors with Proton Beam.
To study the effect of skull surgical clips and bone artifacts on dose distribution using proton beams for treatment planning. Proton treatment plans with double scattering technique for ten patients were generated on patients with brain tumors having had prior surgery with skull bone surgical clips. Appropriate clinical, dosimetric and geometric margins were added for each tumor volume and dose was prescribed to clinical target volume (CTV). The CT images used for treatment planning show streaking artifacts due to the clips and there are also soft artifacts on tissue adjacent to skull bone due to high density of bone. We performed two plans for each patient; a plan without correcting for artifacts and a second plan replacing the artifacts with normal tissue. For the second plan, the artifacts were contoured and a CT numbers obtained by sampling pixels in adjacent areas without artifacts was assigned to the contours. We compared the two plans by evaluating the dose volume histogram (DVH) for tumor coverage and the mean, median, and maximum dose for normal tissue structures. Volume of CTV receiving 99% of the prescribed dose (PD) show a mean difference of 3% higher dose for artifact corrected plans. The normal tissue structures comparison shows and average dose difference of 5%, 8%, 11%, and 30% lower doses for optical chiasm, temporal lobes, brain and optic nerves for artifact corrected plans respectively. In general when using artifact correction for surgical skull clips and skull bone, the tumor coverage tends to be slightly higher. Overall, the normal tissue structures tend to have lower doses depending on the volume, position and the geometry for artifact corrected plans. We recommend the correction of artifacts due to surgical clips and skull bone for brain region proton planning.


Ratiometric Decoding of Pheromones for a Biomimetic Infochemical Communication System.
Biosynthetic infochemical communication is an emerging scientific field employing molecular compounds for information transmission, labelling, and biochemical interfacing; having potential application in diverse areas ranging from pest management to group coordination of swarming robots. Our communication system comprises a chemoemitter module that encodes information by producing volatile pheromone components and a chemoreceiver module that decodes the transmitted ratiometric information via polymer-coated piezoelectric Surface Acoustic Wave Resonator (SAWR) sensors. The inspiration for such a system is based on the pheromone-based communication between insects. Ten features are extracted from the SAWR sensor response and analysed using multi-variate classification techniques, i.e., Linear Discriminant Analysis (LDA), Probabilistic Neural Network (PNN), and Multilayer Perception Neural Network (MLPNN) methods, and an optimal feature subset is identified. A combination of steady state and transient features of the sensor signals showed superior performances with LDA and MLPNN. Although MLPNN gave excellent results reaching 100% recognition rate at 400 s, over all time stations PNN gave the best performance based on an expanded data-set with adjacent neighbours. In this case, 100% of the pheromone mixtures were successfully identified just 200 s after they were first injected into the wind tunnel. We believe that this approach can be used for future chemical communication employing simple mixtures of airborne molecules.


High fructose corn syrup and diabetes prevalence: a global perspective.
The overall aim of this study was to evaluate, from a global and ecological perspective, the relationships between availability of high fructose corn syrup (HFCS) and prevalence of type 2 diabetes. Using published resources, country-level estimates (n =43 countries) were obtained for: total sugar, HFCS and total calorie availability, obesity, two separate prevalence estimates for diabetes, prevalence estimate for impaired glucose tolerance and fasting plasma glucose. Pearson's correlations and partial correlations were conducted in order to explore associations between dietary availability and obesity and diabetes prevalence. Diabetes prevalence was 20% higher in countries with higher availability of HFCS compared to countries with low availability, and these differences were retained or strengthened after adjusting for country-level estimates of body mass index (BMI), population and gross domestic product (adjusted diabetes prevalence=8.0 vs. 6.7%, p=0.03; fasting plasma glucose=5.34 vs. 5.22 mmol/L, p=0.03) despite similarities in obesity and total sugar and calorie availability. These results suggest that countries with higher availability of HFCS have a higher prevalence of type 2 diabetes independent of obesity.


Ipriflavone inhibits osteolytic bone metastasis of human breast cancer cells in a nude mouse model.
Osteolytic bone metastasis is a frequent problem in the treatment of cancer. Ipriflavone, a synthetic isoflavone that inhibits osteoclastic bone resorption, has been used for the treatment of osteoporosis in some countries. Some other isoflavones also exhibit an antitumor effect in vitro and in vivo. Here, we studied the effects of ipriflavone on osteolytic bone metastasis of MDA-231 human breast cancer cells injected intracardially into athymic nude mice (ICR-nu/nu). Daily oral administration of ipriflavone at 12 mg/mouse significantly inhibited the development of new osteolytic bone metastases (p < 0.05) and the progression of established osteolytic lesions (p = 0.01), prolonging the life of tumor-bearing mice (p = 0.01 vs. control). In addition, ipriflavone reduced the number of osteoclasts at the bone-cancer interface with no severe adverse effects on the host. In vitro, ipriflavone inhibited the proliferation and DNA synthesis of MDA-231 cells and blocked the ligand-induced phosphorylation of Tyr(845) of the EGFR. Ipriflavone did not promote apoptosis of MDA-231 cells. Our results show that ipriflavone not only directly inhibits the growth of cancer cells but also reduces osteoclasts to prevent the soft tissue tumor burden and osteolytic bone metastases. These findings raise the possibility that ipriflavone may be of use as a therapeutic agent against osteolytic bone metastasis.


Quantitative bacterial counts in the bone marrow of Vietnamese patients with typhoid fever.
Bone marrow culture (BMC) is the reference standard for typhoid fever diagnosis. We studied the additional yield of BMC over blood culture (BC) and the relationship between quantitative BMC counts and severe disease. Hospitalised Vietnamese patients with suspected typhoid fever were prospectively investigated with a BC, BMC, faecal culture and quantitative BMC counts. Salmonella typhi was isolated in 195 of 231 patients: from BC and BMC in 144 (73.8%), from BMC alone in 33 (16.9%), from BC alone in 12 (6.2%) and from faeces alone in 6 (3.1%). In 167 patients the median extracellular count of S. typhi was 2.5 cfu/mL (interquartile range [IQR] 0-10) and the intracellular count was 10.5 cfu/mL (IQR 2-42) with a ratio of 1.3 bacteria/cell (IQR 0.6-2.5). The median count of intracellular bacteria in 24 patients with severe disease was 46 bacteria/cell (IQR 9-105) compared with 6.5 bacteria/cell (IQR 2-34) in 143 with non-severe disease (p=0.005). The intracellular BMC count was negatively correlated with the peripheral white cell count and positively correlated with hepatomegaly, splenomegaly, aspartate transaminase, a positive BC and the fever clearance time following treatment with azithromycin, ofloxacin or a combination of the two. BMC gave a moderate additional yield over BC. Intracellular BMC counts may reflect the bacterial load in typhoid fever.


Network analysis of social changes in a captive chimpanzee community following the successful integration of two adult groups.
Chimpanzees are highly territorial and have the potential to be extremely aggressive toward unfamiliar individuals. In the wild, transfer between groups is almost exclusively completed by nulliparous females, yet in captivity there is often a need to introduce and integrate a range of individuals, including adult males. We describe the process of successfully integrating two groups of chimpanzees, each containing 11 individuals, in the Budongo Trail facility at the Royal Zoological Society of Scotland's Edinburgh Zoo. We use social network analysis to document changes in group dynamics within this population over the 16 months following integration. Aggression rates were low overall and members of the two original groups engaged in significantly fewer aggressive interactions over time. Association and grooming data indicate that relationships between members of the original groups became stronger and more affiliative with time. Despite these positive indicators the association data revealed the continued existence of two distinct subgroups, a year after integration. Our data show that when given complex space and freedom to exhibit natural fission-fusion groupings, in which the chimpanzees choose whom they wish to associate and interact with, the building of strong affiliative relationships with unfamiliar individuals is a very gradual process.


Differentially expressed proteins in nitric oxide-stimulated NIH/3T3 fibroblasts: implications for inhibiting cancer development.
Recent evidence shows that nitric oxide (NO) may exhibit both pro-cancer and anti-cancer activities. The present study aimed to determine the differentially expressed proteins in NO-treated NIH/3T3 fibroblasts in order to investigate whether NO induces proteins with pro-cancer or anti-cancer effects. The cells were treated with 300 μM of an NO donor 3,3-bis-(aminoethyl)-1-hydroxy-2-oxo-1-triazene (NOC-18) for 12 h. The changed protein patterns, which were separated by two-dimensional electrophoresis using pH gradients of 4-7, were conclusively identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis of the peptide digests. Seventeen differentially expressed proteins were identified in NOC-18-treated cells. Nine proteins [vinculin protein, keratin 19, ubiquitous tropomodulin, F-actin capping protein (α1 subunit), tropomyosin 3, 26S proteasome-associated pad1 homolog, T-complex protein 1 (ε subunit) N(G)-dimethylarginine dimethylaminohydrolase, and heat shock protein 90] were increased and eight proteins (heat shock protein 70, glucosidase II, lamin B1, calreticulin, nucleophosmin 1, microtubule-associated protein retinitis pigmentosa/end binding family member 1, 150 kD oxygen-regulated protein precursor, and heat shock 70-related protein albino or pale green 2) were decreased by NOC-18 in the cells. Thirteen proteins are related to the suppression of cancer cell proliferation, invasion, and metastasis while two proteins (heat shock protein 90 and N(G)-dimethylarginine dimethylaminohydrolase) are related to carcinogenesis. The functions of 150 kD oxygen-regulated protein precursor and T-complex protein 1 (ε subunit) are unknown in relation to carcinogenesis. Most proteins differentially expressed by NOC-18 are involved in inhibiting cancer development.


Tumor-Stroma Interactions in Bone Metastasis: Molecular Mechanisms and Therapeutic Implications.
Metastasis and associated complications are the major cause of death for cancer patients. The incidence of bone metastasis is among the highest in cancers arising from breast, prostate, and lung. Common skeletal-related events caused by bone metastasis include aberrant bone remodeling (osteolytic, osteoblastic, and mixed), bone pain, fracture, spinal cord compression, and life-threatening hypercalcemia. It is now known that interactions between tumor cells and bone stroma lie at the core of major steps of bone-metastasis progression. Approved pharmaceutical drugs for the treatment of bone metastasis, including bisphosphonate and denosumab, were designed to target bone stromal cell components. In recent years, research in our laboratory and others has revealed intricate tumor-stroma interactions as the driving force behind osteolytic bone-metastasis development, providing a set of new candidates for future drug development. Moreover, recent studies also indicate existence of distinct bone niches in supporting hematopoietic stem cell renewal and differentiation. These niche components are likely utilized by metastatic cancer cells for seeding, progression, and therapy resistance of bone metastasis. Future studies in this direction may discover additional therapeutic targets for bone-metastasis treatment.


Pancreatitis associated with atypical antipsychotics: from the Food and Drug Administration's MedWatch surveillance system and published reports.
To investigate the relative numbers and clinical characteristics of pancreatitis in patients treated with the atypical antipsychotic agents, clozapine, olanzapine, and risperidone, versus the conventional neuroleptic, haloperidol. Pharmacovigilance study of pooled, spontaneously reported adverse events. Government-affiliated drug evaluation center. One hundred ninety-two patients who developed pancreatitis during treatment with one or more antipsychotic agents. Patients were identified with the Food and Drug Administration's MedWatch surveillance program and a MEDLINE search. Most cases of pancreatitis occurred within 6 months after the start of therapy with one or more antipsychotic agents. Of the reports of pancreatitis occurring in conjunction with these drugs, 40%, 33%, 16%, and 12% were in patients receiving treatment with clozapine, olanzapine, risperidone, and haloperidol, respectively. In 50% of the patients receiving haloperidol, an atypical antipsychotic was listed as a concomitant drug. Valproate was administered concomitantly in 23% of patients. Hyperglycemia and acidosis, although uncommon, developed with all the drugs except haloperidol. Twenty-two patients died. In contrast to patients who developed pancreatitis while receiving an atypical antipsychotic, those who developed the disease while receiving haloperidol were women and tended to be older. The number of reports involving the three atypical antipsychotic agents and the relative paucity of reports involving haloperidol, despite its more extensive patient exposure, suggest that atypical antipsychotics may precipitate pancreatitis. However, the risk may not be the same with all agents; pancreatitis was reported most frequently with clozapine, followed by olanzapine, and then risperidone. The temporal relationship of the onset of pancreatitis with the start of drug therapy further supports a cause-and-effect relationship.


Frequency of subclinical herpes zoster in pediatric hematology-oncology patients receiving chemotherapy: A retrospective cohort analysis.
Varicella-zoster virus (VZV) reactivation from the enteric nervous system can cause ileus (Ogilvie's syndrome) in adult patients. Since no pediatric cases have been described, we sought to retrospectively analyze VZV reactivation in pediatric hematology-oncology patients to determine whether VZV infection including subclinical VZV reactivation can induce gastrointestinal complications such as Ogilvie's syndrome. Thirty-five patients who received chemotherapy at our institution between September 2013 and June 2018 were included. Serum samples were collected weekly during hospitalization and every 3 months during outpatient maintenance chemotherapy. A real-time polymerase chain reaction assay was used to measure VZV DNA load in serum. The clinical features of patients with VZV infection were retrospectively analyzed. Of 1165 serum samples, 7 (0.6%) were positive for VZV DNA. VZV DNA was detected in 3 of 35 patients. In patient A, VZV DNA was detected during two episodes. The first episode involved varicella-like eruptions caused by the Oka VZV vaccine strain. The second episode involved herpes zoster (HZ) caused by the same strain. Patients B and C had a clinical course that was typical for HZ caused by wild-type VZV. No gastrointestinal symptoms were observed at the time of VZV infection in these three patients. VZV DNA was not detected in any other samples. No pediatric cases with Ogilvie's syndrome caused by VZV reactivation were demonstrated in this cohort. Additionally, no subclinical VZV reactivation was found in this cohort. Further study is needed to elucidate the precise incidence of pediatric Ogilvie's syndrome caused by VZV reactivation.


Detection of myocardial disorders by magnetic resonance spectroscopy.
Magnetic resonance spectroscopy (MRS) utilizes magnetic resonance signals from nuclei, such as phosphorus-31, to provide information regarding the biochemical composition and metabolic state of cardiac muscle. This technique is the only method available for noninvasive assessment of cardiac metabolism without the need for the application of external radioactive tracers. MRS provides insights into the role of cardiac energetics in ischemic heart disease, heart failure, hypertrophy, and valve disease. Furthermore, response to therapeutic intervention can be monitored using this method. At present, this technique is used as a research tool, because low spatial and temporal resolution, as well as low reproducibility, precludes its diagnostic use in clinical practice; however, higher-field magnetic resonance systems-using, for example, 7 T-will enable improvements in resolution and reproducibility that may take cardiac MRS into the clinical realm.


Independent estimation of the frequency of rare CNVs in the UK population confirms their role in schizophrenia.
Several large, rare chromosomal copy number variants (CNVs) have recently been shown to increase risk for schizophrenia and other neuropsychiatric disorders including autism, ADHD, learning difficulties and epilepsy. We wanted to examine the frequencies of these schizophrenia-associated variants in a large sample of individuals with non-psychiatric illnesses to better understand the robustness and specificity of the association with schizophrenia. We used Affymetrix 500K microarray data from 10,259 individuals from the UK Wellcome Trust Case Control Consortium (WTCCC) who are affected with six non-psychiatric disorders (coronary artery disease, Crohn's disease, hypertension, rheumatoid arthritis, types 1 and 2 diabetes) to establish the frequencies of nine CNV loci strongly implicated in schizophrenia, and compared them with the previous findings. Deletions at 1q21.1, 3q29, 15q11.2, 15q13.1 and 22q11.2 (VCFS region), and duplications at 16p11.2 were found significantly more often in schizophrenia cases, compared with the WTCCC reference set. Deletions at 17p12 and 17q12, were also more common in schizophrenia cases but not significantly so, while duplications at 16p13.1 were found at nearly the same rate as in previous schizophrenia samples. The frequencies of CNVs in the WTCCC non-psychiatric controls at three of the loci (15q11.2, 16p13.1 and 17p12) were significantly higher than those reported in previous control populations. The evidence for association with schizophrenia is compelling for six rare CNV loci, while the remaining three require further replication in large studies. Risk at these loci extends to other neurodevelopmental disorders but their involvement in common non-psychiatric disorders should also be investigated.


Visual Contrast Enhancement Algorithm Based on Histogram Equalization.
Image enhancement techniques primarily improve the contrast of an image to lend it a better appearance. One of the popular enhancement methods is histogram equalization (HE) because of its simplicity and effectiveness. However, it is rarely applied to consumer electronics products because it can cause excessive contrast enhancement and feature loss problems. These problems make the images processed by HE look unnatural and introduce unwanted artifacts in them. In this study, a visual contrast enhancement algorithm (VCEA) based on HE is proposed. VCEA considers the requirements of the human visual perception in order to address the drawbacks of HE. It effectively solves the excessive contrast enhancement problem by adjusting the spaces between two adjacent gray values of the HE histogram. In addition, VCEA reduces the effects of the feature loss problem by using the obtained spaces. Furthermore, VCEA enhances the detailed textures of an image to generate an enhanced image with better visual quality. Experimental results show that images obtained by applying VCEA have higher contrast and are more suited to human visual perception than those processed by HE and other HE-based methods.


Pseudohyperglycemia: Effects of Unwashed Hand after Fruit Peeling or Handling on Fingertips Blood Glucose Monitoring Results.
Self-monitoring of blood glucose (SMBG) is an important component of management for diabetes mellitus (DM), especially in T1DM and T2DM patients who are on insulin therapy. Adequate blood glucose monitoring and prompt intervention are necessary to prevent blood glucose (BG) fluctuation and delay long-term diabetes complications. People with DM are advised to clean their hands before SMBG to remove any dirt or food residue that might affect the reading. The study tested the hypothesis that falsely elevated BG levels from fingertip occur after peeling or handling fruits in an unwashed hand. Fifty apparently healthy nondiabetes volunteers were enrolled. Capillary BG samples were collected from the fingertips after peeling or handling apple, orange, banana, watermelon, and pawpaw, followed by no hand washing for 1 h, cleaning the fingertip with alcohol swab once, five times, and washing hand thoroughly with tap water and drying. These samples were then analyzed with two different glucose meters. The mean BG values, measured from fingertip blood samples after peeling, and handling any of the fruits followed by no hand washing were significantly high, even after cleaning fingertip with a swab of alcohol once. However, there were no significant difference in BG levels measured after peeling and handling fruits followed by hand washing and the level of BG before peeling and handling fruits. Handling of peeled fruits with no hand washing with tap water is associated with overestimation of capillary BG (Pseudohyperglycemia) monitored with glucose meters.


Suggested indications of clinical practice guideline for stem cell-therapy in cardiovascular diseases: A stepwise appropriate use criteria for regeneration therapy.
Despite astonishing progress concerning cardiovascular diseases, patients are still suffering from complications of acute insults. Due to reverse remodeling and improper myocyte rebuilding, heart failure has become a common problem these days which needs more powerful myocardial reconstructing strategies. Indeed, no option cases afflicted with non-healing peripheral vascular diseases; refractory stable and unstable angina is the other field with paucity of proper treatments. For these cases, stem cell-based therapies became optimistic treatment, but lack of guideline-based indications regarding stem-cell is still a major problem which limits application of these cells for such end-stage cases. Here, an outline of appropriateness criteria for stem cell-based therapy is suggested.


How strong is the evidence that conservative treatment reduces pain and improves function in individuals with patellar tendinopathy? A systematic review of randomised controlled trials including GRADE recommendations.
To determine the effectiveness of conservative treatment (CT) on pain and function in patients with patellar tendinopathy (PT) compared with minimal intervention (MI) or other invasive intervention, or in addition to decline eccentric squat. Searches were performed in MEDLINE, Embase, Cochrane, PEDro, SPORTDiscus, CINAHL and AMED databases. All randomised trials that evaluated CT (any intervention not involving invasive procedures or medication) in individuals with PT were included. Two reviewers screened studies, extracted data and assessed risk of bias of all included studies. Where suitable, meta-analyses were conducted; we assessed certainty of the evidence using GRADE methodology. When compared with MI, CT did not improve pain (weighted mean difference (WMD) -2.6, 95% CI -6.5 to 1.2) or function (WMD 1.8, 95% CI -2.4 to 6.1) in the short-term (up to 3 months) follow-up. When compared with invasive intervention, CT did not improve pain (WMD 0.7, 95% CI -0.1 to 1.4) or function (WMD -6.6, 95% CI -13.3 to 0.2) in the short-term follow-up. No overall effects were found for combined CT (when a conservative intervention was added to decline eccentric squat) on pain (WMD -0.5, 95% CI -1.4 to 0.4) or function (WMD -2.3, 95 % -9.1 to 4.6) at short-term follow-up. Single studies showed an effect on pain with iontophoresis at short-term follow-up (d = 2.42) or dry needling at medium/long-term follow-up (d = 1.17) and function with exercise intervention at medium/long-term follow-up (over 3 months) (d = 0.83). Our estimates of treatment effect have only low to very low certainty evidence to support them. This field of sports medicine/sports physiotherapy urgently needs larger, high-quality studies with pain and function among the potential primary outcomes.


Proteolysis of the proofreading subunit controls the assembly of Escherichia coli DNA polymerase III catalytic core.
The C-terminal region of the proofreading subunit (epsilon) of Escherichia coli DNA polymerase III is shown here to be labile and to contain the residues (identified between F187 and R213) responsible for association with the polymerase subunit (alpha). We also identify two alpha-helices of the polymerase subunit (comprising the residues E311-M335 and G339-D353, respectively) as the determinants of binding to epsilon. The C-terminal region of epsilon is degraded by the ClpP protease assisted by the GroL molecular chaperone, while other factors control the overall concentration in vivo of epsilon. Among these factors, the chaperone DnaK is of primary importance for preserving the integrity of epsilon. Remarkably, inactivation of DnaK confers to Escherichia coli inviable phenotype at 42 degrees C, and viability can be restored over-expressing epsilon. Altogether, our observations indicate that the association between epsilon and alpha subunits of DNA polymerase III depends on small portions of both proteins, the association of which is controlled by proteolysis of epsilon. Accordingly, the factors catalysing (ClpP, GroL) or preventing (DnaK) this proteolysis exert a crucial checkpoint of the assembly of Escherichia coli DNA polymerase III core.


The role of Helicobacter pylori in urticaria and atopic dermatitis.
Helicobacter pylori, a common cause of gastritis and peptic ulcer, has been associated with several extragastrointestinal diseases. Many studies have shown a positive relation between H. pylori infection and both chronic idiopathic urticaria and atopic dermatitis. The study included 20 patients diagnosed with chronic idiopathic urticaria and 20 with atopic dermatitis. A randomized sample of 20 healthy individuals was selected as a control group. H. pylori infection was assessed using the C-urea breath test, and anti-H. pylori IgG antibody titers were determined by enzyme-linked immunosorbent assay. In the chronic idiopathic urticaria group, the urea breath test titer was positive in 75% of patients, and anti-IgG antibodies were also detected in 75% of patients. In the atopic dermatitis group, titer was positive in 70% and antibodies were detected in 65% of patients, while in the control group, the urea breath test titer was positive in 55% and antibodies were seen in 20% of patients. This difference was statistically highly significant (p < 0.001) in the case of anti-H. pylori antibodies in relation to chronic idiopathic urticaria and significant (p < 0.05) in the case of atopic dermatitis. These results provide some evidence of a relationship between H. pylori infection and both chronic idiopathic urticaria and atopic dermatitis. Treatment of infection demonstrated by reduction in C-urea breath test and anti-H. pylori antibody titers resulted in partial improvement of clinical symptoms in patients with atopic dermatitis.


Development of spontaneous multisystem autoimmune disease and hypersensitivity to antibody-induced inflammation in Fcgamma receptor IIa-transgenic mice.
The major human Fc receptor, FcgammaRIIa, is the most widespread activating FcR. Our aim was to determine the role of FcgammaRIIa in a transgenic mouse model of immune complex-mediated autoimmunity and to characterize the development of spontaneous autoimmune disease. Arthritis was induced in normal and FcgammaRIIa-transgenic mice by immunization with type II collagen (CII) or by transfer of arthritogenic anti-CII antibodies. Also, mice that spontaneously developed autoimmune disease were assessed by clinical scoring of affected limbs, histology and serology, and measurement of autoantibody titers and cytokine production. FcgammaRIIa-transgenic mice developed collagen-induced arthritis (CIA) more rapidly than did archetypal CIA-sensitive DBA/1 (H-2q) mice, while nontransgenic C57BL/6 (H-2b) mice did not develop CIA when similarly immunized. Passive transfer of a single dose of anti-CII antibody induced a more rapid, severe arthritis in FcgammaRIIa-transgenic mice than in nontransgenic animals. In addition, most immune complex-induced production of tumor necrosis factor alpha by activated macrophages occurred via FcgammaRIIa, not the endogenous mouse FcR. A spontaneous, multisystem autoimmune disease developed in aging (>20 weeks) transgenic mice (n = 25), with a 32% incidence of arthritis, and by 45 weeks, all mice had developed glomerulonephritis and pneumonitis, and most had antihistone antibodies. Elevated IgG2a levels were seen in mice with CIA and in those with spontaneous disease. The presence of enhanced passive and induced autoimmunity, as well as the emergence of spontaneous autoimmune disease at 20-45 weeks of age, suggest that FcgammaRIIa is a very important factor in the pathogenesis of autoimmune inflammation and a possible target for therapeutic intervention.


Adjustment of speaker's referential expressions to an addressee's likely knowledge and link with theory of mind abilities.
To communicate cooperatively, speakers must determine what constitutes the common ground with their addressee and adapt their referential choices accordingly. Assessing another person's knowledge requires a social cognition ability termed theory of mind (ToM). This study relies on a novel referential communication task requiring probabilistic inferences of the knowledge already held by an addressee prior to the study. Forty participants were asked to present 10 movie characters and the addressee, who had the same characters in a random order, was asked to place them in order. ToM and other aspects of social cognition were also assessed. Participants used more information when presenting likely unknown than likely known movie characters. They particularly increased their use of physical descriptors, which most often accompanied movie-related information. Interestingly, a significant relationship emerged between our ToM test and the increased amount of information given for the likely unknown characters. These results suggest that speakers use ToM to infer their addressee's likely knowledge and accordingly adapt their referential expressions.


The impact of a laser-microtextured collar on crestal bone level and clinical parameters under various placement and loading protocols.
Physical attachment of connective tissue fibers to a laser-microtextured (8- and 12-μm grooves) surface on the collar of an implant has been demonstrated using human histology. Related clinical research has suggested that this microtextured surface may help to decrease initial bone loss after implant placement. The aim of this retrospective study was to compare crestal bone heights and clinical parameters between implants with laser-microtextured and machined collars placed and loaded with different protocols. This study evaluated 300 single implants in 300 patients (155 men, 145 women; mean age: 49.3 years; range: 45 to 75 years). One hundred sixty implants with laser-microtextured collars (L) and 140 with machined collars (M) were used. Implants were grouped into the treatment categories of immediate placement, delayed placement, immediate nonocclusal loading, and delayed loading. For all groups, crestal bone level, attachment level (CAL), Plaque Index, and bleeding on probing were recorded at baseline and 6, 12, and 24 months after loading with the definitive restoration. Nine implants were lost (four L and five M). The type of implant and timing of placement and loading showed no significant influence on survival rates. A mean CAL loss of 1.12 mm was observed during the first 2 years in the M group, while the mean CAL loss observed in the L group was 0.55 mm. Radiographically, L group implants showed a mean crestal bone loss of 0.58 mm, compared to 1.09 mm for the M group. A laser-microtextured surface on the implant collar may mitigate the negative sequelae associated with peri-implant bone loss, regardless of the placement and loading protocols used.


Network-Thinking: Graphs to Analyze Microbial Complexity and Evolution.
The tree model and tree-based methods have played a major, fruitful role in evolutionary studies. However, with the increasing realization of the quantitative and qualitative importance of reticulate evolutionary processes, affecting all levels of biological organization, complementary network-based models and methods are now flourishing, inviting evolutionary biology to experience a network-thinking era. We show how relatively recent comers in this field of study, that is, sequence-similarity networks, genome networks, and gene families-genomes bipartite graphs, already allow for a significantly enhanced usage of molecular datasets in comparative studies. Analyses of these networks provide tools for tackling a multitude of complex phenomena, including the evolution of gene transfer, composite genes and genomes, evolutionary transitions, and holobionts.


Tolerance of entomopathogenic fungi to ultraviolet radiation: a review on screening of strains and their formulation.
Ultraviolet radiation from sunlight is probably the most detrimental environmental factor affecting the viability of entomopathogenic fungi applied to solar-exposed sites (e.g., leaves) for pest control. Most entomopathogenic fungi are sensitive to UV radiation, but there is great inter- and intraspecies variability in susceptibility to UV. This variability may reflect natural adaptations of isolates to their different environmental conditions. Selecting strains with outstanding natural tolerance to UV is considered as an important step to identify promising biological control agents. However, reports on tolerance among the isolates used to date must be analyzed carefully due to considerable variations in the methods used to garner the data. The current review presents tables listing many studies in which different methods were applied to check natural and enhanced tolerance to UV stress of numerous entomopathogenic fungi, including several well-known isolates of these fungi. The assessment of UV tolerance is usually conducted with conidia using dose-response methods, wherein the UV dose is calculated simply by multiplying the total irradiance by the period (time) of exposure. Although irradiation from lamps seldom presents an environmentally realistic spectral distribution, laboratory tests circumvent the uncontrollable circumstances associated with field assays. Most attempts to increase field persistence of microbial agents have included formulating conidia with UV protectants; however, in many cases, field efficacy of formulated fungi is still not fully adequate for dependable pest control.


Confidence of probabilistic predictions modulates the cortical response to pain.
Pain typically evolves over time, and the brain needs to learn this temporal evolution to predict how pain is likely to change in the future and orient behavior. This process is termed temporal statistical learning (TSL). Recently, it has been shown that TSL for pain sequences can be achieved using optimal Bayesian inference, which is encoded in somatosensory processing regions. Here, we investigate whether the confidence of these probabilistic predictions modulates the EEG response to noxious stimuli, using a TSL task. Confidence measures the uncertainty about the probabilistic prediction, irrespective of its actual outcome. Bayesian models dictate that the confidence about probabilistic predictions should be integrated with incoming inputs and weight learning, such that it modulates the early components of the EEG responses to noxious stimuli, and this should be captured by a negative correlation: when confidence is higher, the early neural responses are smaller as the brain relies more on expectations/predictions and less on sensory inputs (and vice versa). We show that participants were able to predict the sequence transition probabilities using Bayesian inference, with some forgetting. Then, we find that the confidence of these probabilistic predictions was negatively associated with the amplitude of the N2 and P2 components of the vertex potential: the more confident were participants about their predictions, the smaller the vertex potential. These results confirm key predictions of a Bayesian learning model and clarify the functional significance of the early EEG responses to nociceptive stimuli, as being implicated in confidence-weighted statistical learning.


Pregnancy management and outcome in patients with four different tetrahydrobiopterin disorders.
Inborn errors of tetrahydrobiopterin (BH4) biosynthesis or recycling are a group of very rare neurometabolic diseases. Following growing awareness and improved availability of drug treatment the number of patients with BH4 disorders reaching adulthood is constantly increasing. Pregnancy care of patients with these disorders is therefore a new challenge for clinicians. This retrospective study summarises for the first time clinical and biochemical monitoring data of 16 pregnancies in seven women with different disorders of BH4 metabolism and evaluates treatment regimens before and during pregnancy in relation to the obstetrical outcome and paediatric follow-up. Worsening of pre-existing neurological symptoms or occurrence of new symptoms during pregnancy was not observed in most of the cases. Treatment regimens remained mostly unchanged. Pregnancies were not complicated by disease-specific features. Organ abnormalities, miscarriage, prematurity, IUGR and chromosomal changes were occasionally reported, without showing any association with the standard drug treatment for BH4 deficiencies. Although our data on 16 pregnancies in seven patients did not present any association of standard drug treatment with an increased rate of pregnancy complications, abnormal obstetrical or paediatric outcome, an intensive clinical and biochemical supervision by a multidisciplinary team before, during and after the pregnancy in any BH4 deficiency is essential since available data on pregnancies in patients with BH4 deficiencies is limited.


The iRhom homology domain is indispensable for ADAM17-mediated TNFα and EGF receptor ligand release.
Membrane-tethered signalling proteins such as TNFα and many EGF receptor ligands undergo shedding by the metalloproteinase ADAM17 to get released. The pseudoproteases iRhom1 and iRhom2 are important for the transport, maturation and activity of ADAM17. Yet, the structural and functional requirements to promote the transport of the iRhom-ADAM17 complex have not yet been thoroughly investigated. Utilising in silico and in vitro methods, we here map the conserved iRhom homology domain (IRHD) and provide first insights into its structure and function. By focusing on iRhom2, we identified different structural and functional factors within the IRHD. We found that the structural integrity of the IRHD is a key factor for ADAM17 binding. In addition, we identified a highly conserved motif within an unstructured region of the IRHD, that, when mutated, restricts the transport of the iRhom-ADAM17 complex through the secretory pathway in in vitro, ex vivo and in vivo systems and also increases the half-life of iRhom2 and ADAM17. Furthermore, the disruption of this IRHD motif was also reflected by changes in the yet undescribed interaction profile of iRhom2 with proteins involved in intracellular vesicle transport. Overall, we provide the first insights into the forward trafficking of iRhoms which is critical for TNFα and EGF receptor signalling.


Mucosal concentrations of N-acetyl-5-aminosalicylic acid related to endoscopic activity in ulcerative colitis patients with mesalamine.
5-Aminosalicylic acid (5-ASA) is a fundamental treatment for mild-to-moderate ulcerative colitis (UC). 5-ASA is taken up into the colonic mucosa and metabolized to N-acetyl-5-ASA (Ac-5-ASA). Few studies have assessed whether mucosal 5-ASA and Ac-5-ASA concentrations are associated with endoscopic remission. This study aimed to investigate differences in 5-ASA and Ac-5-ASA concentrations according to endoscopic activity. This single-center, prospective, cross-sectional study was conducted between March 2018 and February 2019. UC patients who were administered with 5-ASA medication for at least 8 weeks before sigmoidoscopy were enrolled. Mucosal 5-ASA and Ac-5-ASA concentrations were measured using liquid chromatography with tandem mass spectrometry. The primary endpoint was defined as the difference in mucosal concentrations of 5-ASA and Ac-5-ASA, according to the Mayo endoscopic subscore (MES). Mucosal concentrations were analyzed in 50 patients. In the sigmoid colon, the median 5-ASA concentration in patients with MES of 0 (17.3 ng/mg) was significantly higher than MES ≥ 1 (6.4 ng/mg) (P = 0.019). The median 5-ASA concentrations in patients with Ulcerative Colitis Endoscopic Index of Severity ≤ 1 (16.4 ng/mg) were also significantly higher than in patients with Ulcerative Colitis Endoscopic Index of Severity ≥ 2 (4.63 ng/mg) (P = 0.047). In the sigmoid colon, the concentration of Ac-5-ASA was higher in patients with MES of 0 (21.2 ng/mg) than in patients with MES ≥ 1 (5.81 ng/mg) (P = 0.022). The present study showed that mucosal Ac-5-ASA concentrations, as well as 5-ASA concentrations, are higher in UC patients with endoscopic remission. Ac-5-ASA may be useful for a biomarker of 5-ASA efficacy.


Population-Based Assessment of a Biomarker-Based Screening Pathway to Aid Diagnosis of Monogenic Diabetes in Young-Onset Patients.
Monogenic diabetes, a young-onset form of diabetes, is often misdiagnosed as type 1 diabetes, resulting in unnecessary treatment with insulin. A screening approach for monogenic diabetes is needed to accurately select suitable patients for expensive diagnostic genetic testing. We used C-peptide and islet autoantibodies, highly sensitive and specific biomarkers for discriminating type 1 from non-type 1 diabetes, in a biomarker screening pathway for monogenic diabetes. We studied patients diagnosed at age 30 years or younger, currently younger than 50 years, in two U.K. regions with existing high detection of monogenic diabetes. The biomarker screening pathway comprised three stages: 1) assessment of endogenous insulin secretion using urinary C-peptide/creatinine ratio (UCPCR); 2) if UCPCR was ≥0.2 nmol/mmol, measurement of GAD and IA2 islet autoantibodies; and 3) if negative for both autoantibodies, molecular genetic diagnostic testing for 35 monogenic diabetes subtypes. A total of 1,407 patients participated (1,365 with no known genetic cause, 34 with monogenic diabetes, and 8 with cystic fibrosis-related diabetes). A total of 386 out of 1,365 (28%) patients had a UCPCR ≥0.2 nmol/mmol, and 216 out of 386 (56%) were negative for GAD and IA2 and underwent molecular genetic testing. Seventeen new cases of monogenic diabetes were diagnosed (8 common Maturity Onset Diabetes of the Young [Sanger sequencing] and 9 rarer causes [next-generation sequencing]) in addition to the 34 known cases (estimated prevalence of 3.6% [51/1,407] [95% CI 2.7-4.7%]). The positive predictive value was 20%, suggesting a 1-in-5 detection rate for the pathway. The negative predictive value was 99.9%. The biomarker screening pathway for monogenic diabetes is an effective, cheap, and easily implemented approach to systematically screening all young-onset patients. The minimum prevalence of monogenic diabetes is 3.6% of patients diagnosed at age 30 years or younger.


International obligations through collective rights: Moving from foreign health assistance to global health governance.
This article analyzes the growing chasm between international power and state responsibility in health rights, proposing an international legal framework for collective rights - rights that can reform international institutions and empower developing states to realize the determinants of health structured by global forces. With longstanding recognition that many developing state governments cannot realize the health of their peoples without international cooperation, scholars have increasingly sought to codify international obligations under the purview of an evolving human right to health, applying this rights-based approach as a foundational framework for reducing global health inequalities through foreign assistance. Yet the inherent limitations of the individual human rights framework stymie the right to health in impacting the global institutions that are most crucial for realizing underlying determinants of health through the strengthening of primary health care systems. Whereas the right to health has been advanced as an individual right to be realized by a state duty-bearer, the authors find that this limited, atomized right has proven insufficient to create accountability for international obligations in global health policy, enabling the deterioration of primary health care systems that lack the ability to address an expanding set of public health claims. For rights scholars to advance disease protection and health promotion through national primary health care systems - creating the international legal obligations necessary to spur development supportive of the public's health - the authors conclude that scholars must look beyond the individual right to health to create collective international legal obligations commensurate with a public health-centered approach to primary health care. Through the development and implementation of these collective health rights, states can address interconnected determinants of health within and across countries, obligating the international community to scale-up primary health care systems in the developing world and thereby reduce public health inequities through global health governance.


Intentional response distortion on personality tests: using eye-tracking to understand response processes when faking.
Intentional response distortion or faking among job applicants completing measures such as personality and integrity tests is a concern in personnel selection. The present study aimed to investigate whether eye-tracking technology can improve our understanding of the response process when faking. In an experimental within-participants design, a Big Five personality test and an integrity measure were administered to 129 university students in 2 conditions: a respond honestly and a faking good instruction. Item responses, response latencies, and eye movements were measured. Results demonstrated that all personality dimensions were fakeable. In support of the theoretical position that faking involves a less cognitively demanding process than responding honestly, we found that response times were on average 0.25 s lower and participants had less eye fixations in the fake good condition [corrected]. However, in the fake good condition, participants had more fixations on the 2 extreme response options of the 5-point answering scale, and they fixated on these more directly after having read the question. These findings support the idea that faking leads to semantic rather than self-referenced item interpretations. Eye-tracking was demonstrated to be potentially useful in detecting faking behavior, improving detecting rates over and beyond response extremity and latency metrics.


Targeting IGF Perturbs Global Replication through Ribonucleotide Reductase Dysfunction.
Inhibition of IGF receptor (IGF1R) delays repair of radiation-induced DNA double-strand breaks (DSB), prompting us to investigate whether IGF1R influences endogenous DNA damage. Here we demonstrate that IGF1R inhibition generates endogenous DNA lesions protected by 53BP1 bodies, indicating under-replicated DNA. In cancer cells, inhibition or depletion of IGF1R delayed replication fork progression accompanied by activation of ATR-CHK1 signaling and the intra-S-phase checkpoint. This phenotype reflected unanticipated regulation of global replication by IGF1 mediated via AKT, MEK/ERK, and JUN to influence expression of ribonucleotide reductase (RNR) subunit RRM2. Consequently, inhibition or depletion of IGF1R downregulated RRM2, compromising RNR function and perturbing dNTP supply. The resulting delay in fork progression and hallmarks of replication stress were rescued by RRM2 overexpression, confirming RRM2 as the critical factor through which IGF1 regulates replication. Suspecting existence of a backup pathway protecting from toxic sequelae of replication stress, targeted compound screens in breast cancer cells identified synergy between IGF inhibition and ATM loss. Reciprocal screens of ATM-proficient/deficient fibroblasts identified an IGF1R inhibitor as the top hit. IGF inhibition selectively compromised growth of ATM-null cells and spheroids and caused regression of ATM-null xenografts. This synthetic-lethal effect reflected conversion of single-stranded lesions in IGF-inhibited cells into toxic DSBs upon ATM inhibition. Overall, these data implicate IGF1R in alleviating replication stress, and the reciprocal IGF:ATM codependence we identify provides an approach to exploit this effect in ATM-deficient cancers. SIGNIFICANCE: This study identifies regulation of ribonucleotide reductase function and dNTP supply by IGFs and demonstrates that IGF axis blockade induces replication stress and reciprocal codependence on ATM. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/81/8/2128/F1.large.jpg.


News coverage of the E-cigarette, or Vaping, product use Associated Lung Injury (EVALI) outbreak and internet searches for vaping cessation.
In the latter half of 2019, an outbreak of pulmonary disease in the USA resulted in 2807 hospitalisations and 68 deaths, as of 18 February 2020. Given the severity of the outbreak, we assessed whether articles during the outbreak era more frequently warned about the dangers of vaping and whether internet searches for vaping cessation increased. Using Tobacco Watcher, a media monitoring platform that automatically identifies and categorises news articles from sources across the globe, we obtained all articles that (a) discussed the outbreak and (b) primarily warned about the dangers of vaping. We obtained internet search trends originating from the USA that mentioned 'quit' or 'stop' and 'e cig(s),' 'ecig(s),' 'e-cig(s),' 'e cigarette(s),' 'e-cigarette(s),' 'electronic cigarette(s),' 'vape(s),' 'vaping' or 'vaper(s)' from Google Trends (eg, 'how do I quit vaping?'). All data were obtained from 1 January 2014 to 18 February 2020 and ARIMA models were used with historical trends to forecast the ratio of observed to expected search volumes during the outbreak era. News of the vaping-induced pulmonary disease outbreak was first reported on 25 July 2019 with 195 articles, culminating in 44 512 articles by 18 February 2020. On average, news articles warning about the dangers of vaping were 130% (95% prediction interval (PI): -15 to 417) and searches for vaping cessation were 76% (95% PI: 28 to 182) higher than expected levels for the days during the period when the sources of the outbreak were unknown (25 July to 27 September 2019). News and searches stabilised just after the US Centers for Disease Control and Prevention reported that a primary source of the outbreak was an additive used in marijuana vapes on 27 September 2019. In sum, there were 12 286 articles archived in Tobacco Watcher primarily warning about the dangers of vaping and 1 025 000 cessation searches following the outbreak. The vaping-induced pulmonary disease outbreak spawned increased coverage about the dangers of vaping and internet searches for vaping cessation. Resources and strategies that respond to this elevated interest should become a priority among public health leaders.


Genome-wide association analysis identifies six new loci associated with forced vital capacity.
Forced vital capacity (FVC), a spirometric measure of pulmonary function, reflects lung volume and is used to diagnose and monitor lung diseases. We performed genome-wide association study meta-analysis of FVC in 52,253 individuals from 26 studies and followed up the top associations in 32,917 additional individuals of European ancestry. We found six new regions associated at genome-wide significance (P < 5 × 10(-8)) with FVC in or near EFEMP1, BMP6, MIR129-2-HSD17B12, PRDM11, WWOX and KCNJ2. Two loci previously associated with spirometric measures (GSTCD and PTCH1) were related to FVC. Newly implicated regions were followed up in samples from African-American, Korean, Chinese and Hispanic individuals. We detected transcripts for all six newly implicated genes in human lung tissue. The new loci may inform mechanisms involved in lung development and the pathogenesis of restrictive lung disease.


FL118, a novel camptothecin analogue, suppressed migration and invasion of human breast cancer cells by inhibiting epithelial-mesenchymal transition via the Wnt/β-catenin signaling pathway.
The aim of the current study was to investigate the effects of FL118, a novel camptothecin analogue, on migration and invasion of human breast cancer cells and the underlying mechanisms of those effects. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and a plate clone formation assay were used to examine inhibition of the proliferation of MDA-MB-231 cells by FL118. Cell cycle distribution was detected using flow cytometry. A wound healing assay and a transwell assay were performed to detect the effects of FL118 on migration and invasion of MDA-MB-231 cells, respectively. qRT-PCR, Western blotting, and immunocytochemistry were used to study the effects of FL118 on expression of epithelial-mesenchymal transition (EMT)-related molecules and Wnt/ β-catenin signaling components in MDA-MB-231 cells. The current results indicated that FL118 inhibited the proliferation, migration and invasion of MDA-MB-231 cells in a dose- and time-dependent manner. FL118 caused MDA-MB-231 cells to accumulate in the S phase. FL118 significantly suppressed the expression of vimentin while enhancing the expression of E-cadherin. Moreover, decreased expression of β-catenin and its targets survivin and cyclin Dl was detected in the nucleus of MDA-MB-231 cells. Taken together, the current results suggest that FL118 inhibited Wnt/β-catenin signaling, leading to suppressed EMT and decreased migration and invasion of breast cancer cells.


Normal hypermutation in antibody genes from congenic mice defective for DNA polymerase iota.
Several low fidelity DNA polymerases participate in generating mutations in immunoglobulin genes. Polymerase eta is clearly involved in the process by causing substitutions of A:T base pairs, whereas polymerase iota has a controversial role. Although the frequency of mutations was decreased in the BL2 cell line deficient for polymerase iota, hypermutation was normal in the 129 strain of mice, which has a natural nonsense mutation in the Poli gene. It is possible that the mice compensated for the defect over time, or that polymerase eta substituted in the absence of polymerase iota. To examine polymerase iota in a genetically defined background, we backcrossed the 129 nonsense mutation to the C57BL/6 strain for six generations. Class switch recombination and hypermutation were studied in these mice and in congenic mice doubly deficient for both polymerases iota and eta. The absence of both polymerases did not affect production of IgG1, indicating that these enzymes are not involved in switch recombination. Poli(-/-F6) mice had the same types of nucleotide substitutions in variable genes as their C57BL/6 counterparts, and mice doubly deficient for polymerases iota and eta had the same mutational spectrum as Polh-/- mice. Thus, polymerase iota did not contribute to the mutational spectra, even in the absence of polymerase eta.


Asymmetric cell division shapes naive and virtual memory T-cell immunity during ageing.
Efficient immune responses rely on heterogeneity, which in CD8+ T cells, amongst other mechanisms, is achieved by asymmetric cell division (ACD). Here we find that ageing, known to negatively impact immune responses, impairs ACD in murine CD8+ T cells, and that this phenotype can be rescued by transient mTOR inhibition. Increased ACD rates in mitotic cells from aged mice restore the expansion and memory potential of their cellular progenies. Further characterization of the composition of CD8+ T cells reveals that virtual memory cells (TVM cells), which accumulate during ageing, have a unique proliferation and metabolic profile, and retain their ability to divide asymmetrically, which correlates with increased memory potential. The opposite is observed for naive CD8+ T cells from aged mice. Our data provide evidence on how ACD modulation contributes to long-term survival and function of T cells during ageing, offering new insights into how the immune system adapts to ageing.


Morphological and genomic characterization of Filobasidiella depauperata: a homothallic sibling species of the pathogenic cryptococcus species complex.
The fungal species Cryptococcus neoformans and Cryptococcus gattii cause respiratory and neurological disease in animals and humans following inhalation of basidiospores or desiccated yeast cells from the environment. Sexual reproduction in C. neoformans and C. gattii is controlled by a bipolar system in which a single mating type locus (MAT) specifies compatibility. These two species are dimorphic, growing as yeast in the asexual stage, and producing hyphae, basidia, and basidiospores during the sexual stage. In contrast, Filobasidiella depauperata, one of the closest related species, grows exclusively as hyphae and it is found in association with decaying insects. Examination of two available strains of F. depauperata showed that the life cycle of this fungal species shares features associated with the unisexual or same-sex mating cycle in C. neoformans. Therefore, F. depauperata may represent a homothallic and possibly an obligately sexual fungal species. RAPD genotyping of 39 randomly isolated progeny from isolate CBS7855 revealed a new genotype pattern in one of the isolated basidiospores progeny, therefore suggesting that the homothallic cycle in F. depauperata could lead to the emergence of new genotypes. Phylogenetic analyses of genes linked to MAT in C. neoformans indicated that two of these genes in F. depauperata, MYO2 and STE20, appear to form a monophyletic clade with the MATa alleles of C. neoformans and C. gattii, and thus these genes may have been recruited to the MAT locus before F. depauperata diverged. Furthermore, the ancestral MATa locus may have undergone accelerated evolution prior to the divergence of the pathogenic Cryptococcus species since several of the genes linked to the MATa locus appear to have a higher number of changes and substitutions than their MATalpha counterparts. Synteny analyses between C. neoformans and F. depauperata showed that genomic regions on other chromosomes displayed conserved gene order. In contrast, the genes linked to the MAT locus of C. neoformans showed a higher number of chromosomal translocations in the genome of F. depauperata. We therefore propose that chromosomal rearrangements appear to be a major force driving speciation and sexual divergence in these closely related pathogenic and saprobic species.


The effects of graded caloric restriction: XII. Comparison of mouse to human impact on cellular senescence in the colon.
Calorie restriction (CR) is an effective strategy to delay the onset and progression of aging phenotypes in a variety of organisms. Several molecular players are involved in the anti-aging effects of CR, but mechanisms of regulation are poorly understood. Cellular senescence-a cellular state of irreversible growth arrest-is considered a basic mechanism of aging. Senescent cells accumulate with age and promote a number of age-related pathologies. Whether environmental conditions such as diet affect the accumulation of cellular senescence with age is still unclear. Here, we show that a number of classical transcriptomic markers of senescent cells are reduced in adult but relatively young mice under CR. Moreover, we demonstrate that such senescence markers are not induced in the colon of middle-age human volunteers under CR in comparison with age-matched volunteers consuming normal Western diets. Our data support the idea that the improvement in health span observed in different organisms under CR might be partly due to a reduction in the number of senescent cells.