SU-E-I-95: Multiscale Normalized Metal Artifact Reduction (MS-NMAR) in Computed Tomography.
Medical x-ray CT devices produce images of high quality; however, the metal artifacts may arise from the adverse effect of metal materials present in the imaged objects. Conventional methods for metal artifact reduction (MAR) substitute the contaminated projection data corresponding to metal traces by specific interpolations in the projection space. Nevertheless, these methods usually introduce new artifacts, because the prior information of the imaged object (i.e., the prior image) was not involved. Therefore, a normalized MAR (NMAR) has further been invented, in which the projection space interpolation was carried out with respect to the ratio (i.e., normalization) of projection raw data to re-projection of the prior image. This feature renders NMAR a very high accuracy. However, the normalization in NMAR is more reasonable for the larger scale contents rather than for the smaller scale details. The proposed MS-NMAR method is a generalization of NMAR with a dedicated multiscale framework. Both projection raw data and re-projection of the prior image ill be decomposed into each scale. The normalization followed by the linear interpolation is performed in each scale, in which the larger the scale is the wider the interval the linear interpolation is operated with. Composite projection data are acquired by summing the projection components in all scales. MS-NMAR corrected image is reconstructed from the composite projection data. Real CT data are used to verify the efficiency of the proposed method, and to compare with conventional MAR and NMAR. The quality of reconstructed image after MAR is evaluated by inspecting the region around the metal material. Both NMAR and MS-NMAR have a better performance as compared with the conventional MAR. Meanwhile, MS-NMAR outperforms NMAR, considering it hinders the productions of new artifacts while reducing the original metal artifacts. This work is partially supported by the US National Institute of Health through grants P50-AG025688 and 2P50AG025688.


Design of a 2D no-flow chamber to monitor hematopoietic stem cells.
Hematopoietic stem cells (HSCs) are the most commonly used cell type in cell-based therapy. However, the investigation of their behavior in vitro has been limited by the difficulty of monitoring these non-adherent cells under classical culture conditions. Indeed, fluid flow moves cells away from the video-recording position and prevents single cell tracking over long periods of time. Here we describe a large array of 2D no-flow chambers allowing the monitoring of single HSCs for several days. The chamber design has been optimized to facilitate manufacturing and routine use. The chip contains a single inlet and 800 chambers. The chamber medium can be renewed by diffusion within a few minutes. This allowed us to stain live human HSCs with fluorescent primary antibodies in order to reveal their stage in the hematopoiesis differentiation pathway. Thus we were able to correlate human HSCs' growth rate, polarization and migration to their differentiation stage.


Cardiac resynchronization therapy in heart failure: recent advances and new insights.
Newer non-pharmacological therapies for heart failure are being evaluated for patients of congestive heart failure (CHF). Mechanical support with left ventricular assist devices and heart transplantation are reserved for the minority of patients who have severely decompensated heart failure. Despite these therapeutic advances, it is generally accepted that current therapies do not adequately address the clinical need of patients with heart failure, and additional strategies are being developed. Cardiac resynchronization therapy (CRT) is a new modality that involves synchronization of ventricular contraction and has shown a lot of promise in managing symptomatic patients of CHF who are on optimal medical therapy and have interventricular conduction delay (IVCD). It has improved exercise tolerance and NYHA functional class in such patients in sinus rhythm and a recent meta-analysis has also shown mortality benefits in CHF. Recently benefits of CRT have also been observed in CHF patients who do not have wide QRS complexes on electrocardiogram (EKG). It has also been shown to benefit drug refractory angina in CHF. Recent studies have also focused on the combined use of CRT and implantable cardioverter defibrillator (ICD) and it has shown encouraging results. Our aim in this descriptive review is to define practice guidelines and to improve clinicians' knowledge of the available published clinical evidence, concentrating on few randomized controlled trials.


Surprising outcomes in cannabinoid CB1/CB2 receptor double knockout mice in two models of ischemia.
We tested the hypothesis that CB1/CB2 receptor double knockout would produce significant increases in infarct size and volume and significant worsening in clinical score, using two mouse models, one of permanent ischemia and one of ischemia/reperfusion. Focal cerebral infarcts were created using either photo induced permanent injury or transient middle cerebral artery occlusion. Infarct volume and motor function were evaluated in cannabinoid receptor 1/cannabinoid receptor 2 double knockout mice. The results surprisingly revealed that CB1/CB2 double knockout mice showed improved outcomes, with the most improvements in the mouse model of permanent ischemia. Although the number of individuals suffering from stroke in the United States and worldwide will continue to grow, therapeutic intervention for treatment following stroke remains frustratingly limited. Both the cannabinoid 1 receptor (CB1R) and the cannabinoid 2 receptor (CB2R) have been studied in relationship to stroke. Deletion of the CB2R has been shown to worsen outcome, while selective CB2R agonists have been demonstrated to be neuroprotective following stroke. Although initial studies of CB1R knockout mice demonstrated increased injury following stroke, indicating that activation of the CB1R was neuroprotective, later studies of selective antagonists of the CB1R also demonstrated a protective effect. Surprisingly the double knockout animals had improved outcome. Since the phenotype of the double knockout is not dramatically changed, significant changes in the contribution of other homeostatic pathways in compensation for the loss of these two important receptors may explain these apparently contradictory results.


Application of the three-component bidirectional reflectance distribution function model to Monte Carlo calculation of spectral effective emissivities of nonisothermal blackbody cavities.
We applied the bidirectional reflectance distribution function (BRDF) model consisting of diffuse, quasi-specular, and glossy components to the Monte Carlo modeling of spectral effective emissivities for nonisothermal cavities. A method for extension of a monochromatic three-component (3C) BRDF model to a continuous spectral range is proposed. The initial data for this method are the BRDFs measured in the plane of incidence at a single wavelength and several incidence angles and directional-hemispherical reflectance measured at one incidence angle within a finite spectral range. We proposed the Monte Carlo algorithm for calculation of spectral effective emissivities for nonisothermal cavities whose internal surface is described by the wavelength-dependent 3C BRDF model. The results obtained for a cylindroconical nonisothermal cavity are discussed and compared with results obtained using the conventional specular-diffuse model.


Fine particulate air pollution and human mortality: 25+ years of cohort studies.
Much of the key epidemiological evidence that long-term exposure to fine particulate matter air pollution (PM2.5) contributes to increased risk of mortality comes from survival studies of cohorts of individuals. Although the first two of these studies, published in the mid-1990s, were highly controversial, much has changed in the last 25 + years. The objectives of this paper are to succinctly compile and summarize the findings of these cohort studies using meta-analytic tools and to address several of the key controversies. Independent reanalysis and substantial extended analysis of the original cohort studies have been conducted and many additional studies using a wide variety of cohorts, including cohorts constructed from public data and leveraging natural experiments have been published. Meta-analytic estimates of the mean of the distribution of effects from cohort studies that are currently available, provide substantial evidence of adverse air pollution associations with all-cause, cardiopulmonary, and lung cancer mortality.


A systematic review and meta-analysis on the safety and efficacy of the frozen elephant trunk technique in aortic arch surgery.
The treatment of complex pathologies of the aortic arch and proximal descending aorta represents a significant challenge for cardiac surgeons. Various surgical techniques and prostheses have been implemented over the past several decades, all with varying degrees of success. The introduction of the frozen elephant trunk (FET) technique facilitates one-stage repair of such pathologies. The present systematic review and meta-analysis aims to assess the safety and efficacy of the FET approach in the current literature. Electronic searches were performed using six databases from their inception to July 2013. Relevant studies utilizing the FET technique were identified. Data were extracted and analyzed according to predefined clinical endpoints. Seventeen studies were identified for inclusion for qualitative and quantitative analyses, all of which were observational reports. Pooled mortality was 8.3%, while stroke and spinal cord injuries were 4.9% and 5.1% respectively. Cardiopulmonary bypass time, myocardial ischemia time, and circulatory arrest time strongly correlated with perioperative mortality in a linear relationship, while moderate correlations between cerebral perfusion time and mortality, and circulatory arrest time and spinal cord injury, were also identified. Five-year survival, reported in five studies, ranged between 63-88%. Overall, results of the present systematic review and meta-analysis suggest that the FET procedure can be performed with acceptable mortality and morbidity risks.


Suppression by Pt of CO adsorption and dissociation and methane formation on Fe5C2(100) surfaces.
To understand the chemical origin of platinum promotion effects on iron based Fischer-Tropsch synthesis catalysts, the effects of Pt on CO adsorption and dissociation as well as surface carbon hydrogenation on the Fe5C2(100) facet with different surface C* contents have been studied using the spin-polarized density functional theory method. CO dissociation initiating from diverse sites was calculated through both direct and H-assisted pathways via the CHO intermediate. On the perfect (100) surface, CO can hardly dissociate, and the surface carbon can be facially hydrogenated to CH4. On the C*-defect and C*-free (100) surfaces, CO can strongly adsorb on the C* vacant sites and direct dissociation is favored to occur. The activity is higher with the decrease of the surface carbon content. When platinum atoms are added on the surfaces, the C*-vacancies have a higher activity for CO dissociation than the new sites generated by Pt adsorption. However, both the CO dissociation and the surface carbon consumption through CH4 formation are hindered. The evolution of surface carbon is predicted to be suppressed by the addition of Pt on the Fe5C2(100) surface.


The majority of inducible DNA repair genes in Mycobacterium tuberculosis are induced independently of RecA.
In many species of bacteria most inducible DNA repair genes are regulated by LexA homologues and are dependent on RecA for induction. We have shown previously by analysing the induction of recA that two mechanisms for the induction of gene expression following DNA damage exist in Mycobacterium tuberculosis. Whereas one of these depends on RecA and LexA in the classical way, the other mechanism is independent of both of these proteins and induction occurs in the absence of RecA. Here we investigate the generality of each of these mechanisms by analysing the global response to DNA damage in both wild-type M. tuberculosis and a recA deletion strain of M. tuberculosis using microarrays. This revealed that the majority of the genes that were induced remained inducible in the recA mutant stain. Of particular note most of the inducible genes with known or predicted functions in DNA repair did not depend on recA for induction. Amongst these are genes involved in nucleotide excision repair, base excision repair, damage reversal and recombination. Thus, it appears that this novel mechanism of gene regulation is important for DNA repair in M. tuberculosis.


Plasma triglyceride level is a risk factor for cardiovascular disease independent of high-density lipoprotein cholesterol level: a meta-analysis of population-based prospective studies.
Despite nearly 40 years of research, the role of plasma triglyceride as a risk factor for cardiovascular disease remains elusive. The objectives of the present study were to quantify the magnitude of the association between triglyceride and cardiovascular disease in the general population, and to determine whether this relationship is independent of high-density lipoprotein (HDL) cholesterol, using the semi-quantitative techniques of metaanalysis. Seventeen studies were selected for the analysis based on published reports of population-based, prospective studies, including 46413 men and 10864 women. To insure comparability, only studies reporting the association between fasting triglyceride levels and incident cardiovascular endpoints were included. Using standard meta-analysis calculations, relative risks (RR) and 95% confidence intervals (CI) were calculated and standardized with respect to a 1 mmol/l increase in triglyceride. Multivariable-adjusted RRs were determined for the six studies in men and two studies in women that reported adjustments for HDL cholesterol. For men and women, the univariate RRs for triglyceride were 1.32 (95% Cl 1.26-1.39) and 1.76 (95% Cl 1.50-2.07), respectively, indicating an approximately 30% increased risk in men and a 75% increase in women. Adjustment of HDL cholesterol and other risk factors attenuated these RRs to 1.14 (95% Cl 1.05-1.28) and 1.37 (95% Cl 1.13-1.66), respectively, which were still statistically significant values. Based on combined data from prospective studies, triglyceride is a risk factor for cardiovascular disease for both men and women in the general population, independent of HDL cholesterol. These finding demonstrate the necessity for clinical trials to evaluate whether lowering plasma triglyceride decreases the risk of cardiovascular disease.


Urinary tract infection in the setting of vesicoureteral reflux.
Vesicoureteral reflux (VUR) is the most common underlying etiology responsible for febrile urinary tract infections (UTIs) or pyelonephritis in children. Along with the morbidity of pyelonephritis, long-term sequelae of recurrent renal infections include renal scarring, proteinuria, and hypertension. Treatment is directed toward the prevention of recurrent infection through use of continuous antibiotic prophylaxis during a period of observation for spontaneous resolution or by surgical correction. In children, bowel and bladder dysfunction (BBD) plays a significant role in the occurrence of UTI and the rate of VUR resolution. Effective treatment of BBD leads to higher rates of spontaneous resolution and decreased risk of UTI.


Thermal capsulorrhaphy in the treatment of multidirectional instability of the shoulder.
The treatment of symptomatic multidirectional instability (MDI) of the shoulder has proven a challenge to orthopaedic surgeons. Patients who refuse activity modification often fail nonsurgical management, and traditional surgical approaches may trade the dysfunction of instability for that of decreased motion and possible future arthrosis. Recently, several methods of arthroscopic thermal capsulorrhaphy have gained popularity. Studies have shown that capsular shrinkage occurs due to the denaturation of type I collagen with maintenance of heat-stable intermolecular cross-links. Thermally treated tissues undergo a predictable healing response. The primary advantages of this procedure are the decreased surgical morbidity and ease of procedure compared with open capsular shift. Additionally, it may provide improved stability and decreased pain while maintaining range of motion. When radiofrequency treatment provides inadequate visualized capsular contracture, arthroscopic rotator interval closure can improve stability without resorting to an open procedure. While initial results in certain clinical series are promising, the long-term efficacy remains to be seen.


Tumor necrosis factor genetic polymorphisms and response to antiviral therapy in patients with chronic hepatitis C.
Hepatitis C virus (HCV) is the major causal agent of non-A, non-B hepatitis and the leading indication for liver transplantation worldwide. The emerging field of immunogenetics has confirmed the significant role of heritability in host immune responses to infectious pathogens. Both the major and non-major histocompatibility complex genes are increasingly identified as candidate genes hypothesized to influence the susceptibility to, or the course of, a particular disease. We hypothesized that polymorphisms within the major histocompatibility complex class III region that encode for tumor necrosis factors (TNF)-alpha and TNF-beta might be predictive of response to antiviral therapy in patients with chronic hepatitis C. A total of 155 subjects, including 110 HCV-seropositive individuals undergoing antiviral therapy and 45 ethnically similar HCV-negative controls, were studied. The HCV-positive patients had undergone antiviral treatment with either interferon monotherapy (n = 73) or in combination with ribavirin (n = 37) and were categorized as either nonresponders, sustained responders, or relapsers. Sixty (55%) patients had genotype 1 (1a or 1b). Genomic DNA was extracted, followed by polymerase chain reaction amplification and sequencing for two promoter TNF-alpha variants (at positions -238 and -308), as well as restriction fragment length analysis for four polymorphic loci within the TNF-beta gene (NcoI, TNFc, aa13, aa26). Although there was a trend toward higher frequency of the A allele in the TNF 238 promoter among HCV-infected patients (12% vs 4%), there were no significant differences in the distribution of the genotypic polymorphisms between patients and controls. Patients with the TNF 238 A allele had higher pretreatment viral loads as compared with patients homozygous for the wild type allele (7.2 x 10(6) +/- 4.2 x 10(6) copies/ml vs 3.8 x 10(6) +/- 0.34 x 10(6) copies/ml, p = 0.03). However, there was no association between TNF genetic markers, including multiple haplotypic combinations, and response to therapy. In addition, there was no correlation with these polymorphic loci and histological severity of liver disease. Although previous work has suggested potential roles for TNF in the pathogenesis of HCV infection, we were unable to identify any link between TNF genetic polymorphisms and histological severity or response to antiviral therapy.


Lateralization in response to social stimuli in a cooperatively breeding cichlid fish.
Cerebral lateralization, an evolutionarily ancient and widespread phenomenon among vertebrates, is thought to bestow cognitive advantages. The advantages of lateralization at the individual-level do not necessarily require that the entire population share the same pattern of lateralization. In fact, directional bias in lateralization may lead to behavioural predictability and enhanced predator success or prey evasion. Recent theory has suggested that population-level lateralization may be favored if individuals are better able to perform coordinated behaviours, providing a distinct advantage in cooperative contexts. Here we test whether the highly social, cooperatively breeding cichlid fish Neolamprologus pulcher shows lateralized responses to a social stimulus. We found population-level biases in males; on average male N. pulcher use their right eye/left hemisphere to view their mirror image. Individual females had a preferred hemisphere, but these preferences appeared not to be directionally aligned among females. We discuss these results in the context of coordinated social behaviour and suggest future research directions.


Functional analysis of gene duplications in Saccharomyces cerevisiae.
Gene duplication can occur on two scales: whole-genome duplications (WGD) and smaller-scale duplications (SSD) involving individual genes or genomic segments. Duplication may result in functionally redundant genes or diverge in function through neofunctionalization or subfunctionalization. The effect of duplication scale on functional evolution has not yet been explored, probably due to the lack of global knowledge of protein function and different times of duplication events. To address this question, we used integrated Bayesian analysis of diverse functional genomic data to accurately evaluate the extent of functional similarity and divergence between paralogs on a global scale. We found that paralogs resulting from the whole-genome duplication are more likely to share interaction partners and biological functions than smaller-scale duplicates, independent of sequence similarity. In addition, WGD paralogs show lower frequency of essential genes and higher synthetic lethality rate, but instead diverge more in expression pattern and upstream regulatory region. Thus, our analysis demonstrates that WGD paralogs generally have similar compensatory functions but diverging expression patterns, suggesting a potential of distinct evolutionary scenarios for paralogs that arose through different duplication mechanisms. Furthermore, by identifying these functional disparities between the two types of duplicates, we reconcile previous disputes on the relationship between sequence divergence and expression divergence or essentiality.


A Human IPS Model Implicates Embryonic B-Myeloid Fate Restriction as Developmental Susceptibility to B Acute Lymphoblastic Leukemia-Associated ETV6-RUNX1.
ETV6-RUNX1 is associated with childhood acute B-lymphoblastic leukemia (cALL) functioning as a first-hit mutation that initiates a clinically silent pre-leukemia in utero. Because lineage commitment hierarchies differ between embryo and adult, and the impact of oncogenes is cell-context dependent, we hypothesized that the childhood affiliation of ETV6-RUNX1 cALL reflects its origins in a progenitor unique to embryonic life. We characterize the first emerging B cells in first-trimester human embryos, identifying a developmentally restricted CD19-IL-7R+ progenitor compartment, which transitions from a myeloid to lymphoid program during ontogeny. This developmental series is recapitulated in differentiating human pluripotent stem cells (hPSCs), thereby providing a model for the initiation of cALL. Genome-engineered hPSCs expressing ETV6-RUNX1 from the endogenous ETV6 locus show expansion of the CD19-IL-7R+ compartment, show a partial block in B lineage commitment, and produce proB cells with aberrant myeloid gene expression signatures and potential: features (collectively) consistent with a pre-leukemic state.


The ever-changing landscape of rotavirus serotypes.
Rotavirus is a double-stranded RNA virus that is characterized by substantial genetic diversity. The various serotypes of rotavirus have been determined by the presence of neutralizing epitopes on the outer capsid of the protein shell. At present, 5 rotavirus serotypes (G1, G2, G3, G4, G9) are the predominant circulating strains, accounting for approximately 95% of strains worldwide, although there is considerable geographic variability. Incidence rates for various serotypes also vary temporally with seasonal and year-to-year fluctuations. Unusual serotypes are generally uncommon, but new serotypes can emerge. In particular, G9[P8], a reassortment virus, was first identified in 1983 and in the last 10 to 15 years has become widely distributed worldwide. Indeed, G9[P8] has become highly prevalent in many countries in Europe and Australia, with somewhat lower incidence rates in South America, Africa, and Asia. The heterogeneity and ever-changing epidemiology of rotavirus underscores the need for continued surveillance to ensure that vaccination programs provide optimal protection.


Functional imaging of stroke recovery: what have we learnt and where do we go from here?
Functional brain imaging techniques have been used to visualise patterns of activity following stroke and to characterise how these patterns change with recovery or rehabilitation. Some consensus is now emerging on patterns that are predictive of improved outcome, and therapeutic strategies are beginning to be guided by such findings. However, patient heterogeneity predicts that the same approach will not be appropriate for all. Future studies should aim to characterise the factors that influence this heterogeneity, and to individualise rehabilitation strategies based in part on early imaging findings. Functional imaging studies of stroke should also embrace recent methodological and conceptual advances that allow for fuller characterisation of the structural and functional properties of distributed brain networks.


Stem cell therapy. Use of differentiated pluripotent stem cells as replacement therapy for treating disease.
Pluripotent stem cells (PSCs) directed to various cell fates holds promise as source material for treating numerous disorders. The availability of precisely differentiated PSC-derived cells will dramatically affect blood component and hematopoietic stem cell therapies and should facilitate treatment of diabetes, some forms of liver disease and neurologic disorders, retinal diseases, and possibly heart disease. Although an unlimited supply of specific cell types is needed, other barriers must be overcome. This review of the state of cell therapies highlights important challenges. Successful cell transplantation will require optimizing the best cell type and site for engraftment, overcoming limitations to cell migration and tissue integration, and occasionally needing to control immunologic reactivity, as well as a number of other challenges. Collaboration among scientists, clinicians, and industry is critical for generating new stem cell-based therapies.


Role of human cripto-1 in tumor angiogenesis.
Human cripto-1 (CR-1) promotes cell transformation and increases migration and invasion of various mouse and human epithelial cell lines. We investigated whether CR-1 also stimulates angiogenesis. We used human umbilical vein endothelial cells (HUVECs) to measure in vitro migration with fibronectin-coated Boyden chambers, invasion with Matrigel-coated Boyden chambers, proliferation with a tetrazolium salt, and differentiation with an in vitro Matrigel assay. We investigated new blood vessel formation in vivo by use of Matrigel-filled silicone cylinders implanted under the skin of nude mice and by use of a breast cancer xenograft model with CR-1-transfected or control Neo-transfected MCF-7 human breast cancer cells. We also used a blocking anti-CR-1 monoclonal antibody to investigate the role of CR-1 in angiogenesis in vivo and in vitro. All statistical tests were two-sided. CR-1 stimulated HUVEC proliferation, migration, and invasion and induced HUVEC differentiation into vascular-like structures on Matrigel. In vivo, recombinant CR-1 protein induced microvessel formation in Matrigel-filled silicone cylinders, and microvessel formation was statistically significantly inhibited with a blocking anti-CR-1 monoclonal antibody (CR-1 and antibody = 127% of microvessel formation compared with that in untreated control cylinders and CR-1 alone = 259%; difference = 132%, 95% confidence interval [CI] = 123% to 140%; P<.001). Tumors formed by CR-1-transfected MCF-7 cells in the cleared mammary fat pad of nude mice had higher microvessel density than tumors formed by control Neo-transfected MCF-7 cells (CR-1-transfected cells = 4.66 vessels per field and Neo-transfected cells = 2.33 vessels per field; difference = 2.33 vessels per field, 95% CI = 1.2 to 2.8; P = .004). CR-1 appears to have an important role in the multistep process of angiogenesis.


Anger and sadness in response to an emotionally neutral film: evidence for age-specific associations with well-being.
When the association between emotion and well-being is being considered, positive emotions usually come to mind. However, negative emotions serve important adaptive functions and particular negative emotions may be especially adaptive at different stages of adult development. We examined the associations between self-reported negative emotions in response to an emotionally neutral, thematically ambiguous film and subjective well-being among 76 young (age 20-29), 73 middle-aged (age 40-49), and 73 older (age 60-69) adults. Results indicated that higher self-reported anger in response to the film was associated with higher well-being for middle-aged adults, but not for young and older adults. Higher self-reported sadness in response to the film was associated with higher well-being for older adults, but not for young and middle-aged adults. These findings were stronger for cognitive well-being (i.e., satisfaction with life) than for affective well-being (i.e., ratio of dispositional positive to negative affect) and were specific to these emotions (not found for self-reported disgust or fear) and to the emotionally neutral film (not found for sad or disgusting films). Results are discussed in terms of the functions that anger and sadness are thought to serve and the control opportunities afforded in midlife and late life that render these functions differentially adaptive.


Identification of neonatal hearing impairment: distortion product otoacoustic emissions during the perinatal period.
1) To describe distortion product otoacoustic emission (DPOAE) levels, noise levels and signal to noise ratios (SNRs) for a wide range of frequencies and two stimulus levels in neonates and infants. 2) To describe the relations between these DPOAE measurements and age, test environment, baby state, and test time. DPOAEs were measured in 2348 well babies without risk indicators, 353 well babies with at least one risk indicator, and 4478 graduates of neonatal intensive care units (NICUs). DPOAE and noise levels were measured at f2 frequencies of 1.0, 1.5, 2.0, 3.0, and 4.0 kHz, and for primary levels (L1/L2) of 65/50 dB SPL and 75/75 dB SPL. Measurement-based stopping rules were used such that a test did not terminate unless the response was at least 3 dB above the mean noise floor + 2 SDs (SNR) for at least four of five test frequencies. The test would terminate, however, if these criteria were not met after 360 sec. Baby state, test environment, and other test factors were captured at the time of each test. DPOAE levels, noise levels and SNRs were similar for well babies without risk indicators, well babies with risk indicators, and NICU graduates. There was a tendency for larger responses at f2 frequencies of 1.5 and 2.0 Hz, compared with 3.0 and 4.0 kHz; however, the noise levels systematically decreased as frequency increased, resulting in the most favorable SNRs at 3.0 and 4.0 kHz. Response levels were least and noise levels highest for an f2 frequency of 1.0 kHz. In addition, test time to achieve automatic stopping criteria was greatest for 1.0 kHz. With the exception of "active/alert" and "crying" babies, baby state had little influence on DPOAE measurements. Additionally, test environment had little impact on these measurements, at least for the environments in which babies were tested in this study. However, the lowest SNRs were observed for infants who were tested in functioning isolettes. Finally, there were some subtle age affects on DPOAE levels, with the infants born most prematurely producing the smallest responses, regardless of age at the time of test. DPOAE measurements in neonates and infants result in robust responses in the vast majority of ears for f2 frequencies of at least 2.0, 3.0 and 4.0 kHz. SNRs decrease as frequency decreases, making the measurements less reliable at 1.0 kHz. When considered along with test time, there may be little justification for including an f2 frequency at 1.0 kHz in newborn screening programs. It would appear that DPOAEs result in reliable measurements when tests are conducted in the environments in which babies typically are found. Finally, these data suggest that babies can be tested in those states of arousal that are most commonly encountered in the perinatal period.


Quality and labeling information of Moringa oleifera products marketed for HIV-infected people in Zimbabwe.
Labeling information and quality of marketed Moringa oleifera products were assessed. Personnel in 60 pharmacies and 11 herbal shops were interviewed about the sources, dosages, indications and counseling information of Moringa oleifera products. Content analysis of written information provided on Moringa oleifera products was also done. Three samples of Moringa from popular sources were acquired to determine heavy metal content and microbial contamination. The results were compared to specified limits in the European and Chinese pharmacopeia, World Health Organization guidelines and Bureau of Indian Standards. Moringa was available as capsules or powder in 73% of the premises. Moringa was recommended for seven different disease conditions. Four different dosage regimens were prescribed. The main references cited for the counseling information were unscientific literature (62%). The selected Moringa samples were contaminated with bacteria and fungi above the European Pharmacopeia specified limits. Escherichia coli and Salmonella species were present in all three samples. All three samples contained arsenic, nickel and cadmium above the permissible limits. Moringa oleifera with variable labeling information and poor microbial and heavy metal quality is widely available in Zimbabwe.


An innovative mobile data collection technology for public health in a field setting.
The Canadian Network for Public Health Intelligence (CNPHI) is a secure, web-based scientific informatics and biosurveillance platform that leverages disparate public health information resources and expertise for the direct benefit of local, regional and national decision makers. CNPHI fosters collaboration and consultation through innovation in disease surveillance, intelligence exchange, research and response to protect, promote and support public health. The objective of this article is to present the CNPHI 'on the go' mobile application, and to discuss preliminary evaluation of the technology. The mobile application is intended to enable rapid mobile data collection using both online and offline modes supporting various stages of surveillance and response through the extension of data collection and analysis to the mobile environment. Two needs assessment meetings were held with stakeholders representing individuals from government, academia and research institutions, to inform the development of the CNPHI "on the go" mobile application. An initial version of the mobile technology (an "app") was developed and piloted by end-users with expertise in the field. Two focused pilots were conducted to test the technology: Pilot 1: 17-7-2017 to 21-11-2017 (6 participants); Pilot 2: 25-7-2017 to 15-9-2017 (2 participants). An initial consultation was held with the project leads to identify data elements for mobile data collection. A custom data collection form was designed using CNPHI's Web Data technology for each pilot, which was then made available through the mobile app. The technology was assessed using feedback received during each pilot as well as through a survey that was conducted at the conclusion of pilots. Pilot participants reported that the mobile technology allowed seamless data collection, data management and rapid information sharing. Participants also reported that the entire process was seamless, simple, efficient, and that fewer steps were required for data collection and management. Further, significant efficiencies were gained by directly entering information using the mobile app without having to transfer handwritten information into an electronic database. An overall positive experience was reported by participants from both pilots. Literature suggests that traditional methods of surveillance and data collection using a paper based methodology pose many challenges such as data loss and duplication, difficulty in managing the database, and lack of timely access to the data. Accurate and rapid access is critical for public health professionals in order to effectively make decisions and respond to public health emergencies. Results show that the CNPHI "on the go" app is well poised to address some of the suggested challenges. A limitation of this study was that sample size for pilot participation was small for capturing overall feedback on the readiness of the technology for integration into regular surveillance activities and response procedures. CNPHI "on the go" is a customizable technology developed within an already thriving collaborative CNPHI platform used by public health professionals, and performs well as a tool for rapid data collection and secure information sharing.


The homing of exogenous hair follicle mesenchymal stem cells into hair follicle niches.
Hair loss is a debilitating condition associated with the depletion of dermal papilla cells (DPCs), which can be replenished by dermal sheath cells (DSCs). Hence, strategies aimed at increasing the population of DPCs and DSCs hold great promise for the treatment of hair loss. In this study, we demonstrated that introducing exogenous DPCs and DSCs (hair follicle mesenchymal stem cells) could effectively migrate and integrate into the dermal papilla and dermal sheath niches, leading to enhanced hair growth and prolonged anagen phases. However, the homing rates of DPCs and DSCs were influenced by various factors, including recipient mouse depilation, cell passage number, cell dose, and immune rejection. Through in vitro and in vivo experiments, we further discovered that the CXCL13/CXCR5 pathway mediated the homing of DPCs and DSCs into hair follicle niches. This study underscores the potential of cell-based therapies for hair loss by targeted delivery of DPCs and DSCs to their respective niches, and sheds light on the intriguing concept that isolated mesenchymal stem cells can home back to their original niche microenvironment.


Comparative genomics yields insights into niche adaptation of plant vascular wilt pathogens.
The vascular wilt fungi Verticillium dahliae and V. albo-atrum infect over 200 plant species, causing billions of dollars in annual crop losses. The characteristic wilt symptoms are a result of colonization and proliferation of the pathogens in the xylem vessels, which undergo fluctuations in osmolarity. To gain insights into the mechanisms that confer the organisms' pathogenicity and enable them to proliferate in the unique ecological niche of the plant vascular system, we sequenced the genomes of V. dahliae and V. albo-atrum and compared them to each other, and to the genome of Fusarium oxysporum, another fungal wilt pathogen. Our analyses identified a set of proteins that are shared among all three wilt pathogens, and present in few other fungal species. One of these is a homolog of a bacterial glucosyltransferase that synthesizes virulence-related osmoregulated periplasmic glucans in bacteria. Pathogenicity tests of the corresponding V. dahliae glucosyltransferase gene deletion mutants indicate that the gene is required for full virulence in the Australian tobacco species Nicotiana benthamiana. Compared to other fungi, the two sequenced Verticillium genomes encode more pectin-degrading enzymes and other carbohydrate-active enzymes, suggesting an extraordinary capacity to degrade plant pectin barricades. The high level of synteny between the two Verticillium assemblies highlighted four flexible genomic islands in V. dahliae that are enriched for transposable elements, and contain duplicated genes and genes that are important in signaling/transcriptional regulation and iron/lipid metabolism. Coupled with an enhanced capacity to degrade plant materials, these genomic islands may contribute to the expanded genetic diversity and virulence of V. dahliae, the primary causal agent of Verticillium wilts. Significantly, our study reveals insights into the genetic mechanisms of niche adaptation of fungal wilt pathogens, advances our understanding of the evolution and development of their pathogenesis, and sheds light on potential avenues for the development of novel disease management strategies to combat destructive wilt diseases.


[Knowledge level for prescription of nonsteroidal anti-inflammatory by dentists in São José dos Campos, São Paulo State, Brazil].
The abusive use of drugs in clinical routine of dentistry represents a problem of public health. Studies have demonstrated that the effectiveness of anti-hypertensive drugs was decreased by the simultaneous use of Nonsteroidal Anti-Inflammatory Drugs (NAIDs), mainly in elderly patients with primary hypertension. This study was to evaluate the scientific knowledge of the dentists working at the public health service in São José dos Campos, São Paulo, Brazil. This research characterized the scientific upgrade of the dentists concerning the prescription of NAIDs to hypertensive patients. The methods included a questionnaire filled up by the 92 dentists of the public health service in São José dos Campos. The results were statistically analyzed by the Wald and Fisher Tests to evaluate the probability of statistical associations between the studied issues (p-value<0.05). The results showed that the professional knowledge for prescription of NAIDs for hypertensive patients was insufficient. The pharmacological interactions between NAIDs and anti-hypertensive drugs were almost totally unknown. The conclusion was that the update courses in dentistry skills do not offer enough knowledge for pharmacological practices in dentistry.


An evolutionarily stable strategy model for the evolution of dimorphic development in the butterfly Maculinea rebeli, a social parasite of Myrmica ant colonies.
Caterpillars of the butterfly Maculinea rebeli develop as parasites inside ant colonies. In intensively studied French populations, about 25% of caterpillars mature within 1 year (fast-developing larvae [FDL]) and the others after 2 years (slow-developing larvae [SDL]); all available evidence indicates that this ratio is under the control of egg-laying females. We present an analytical model to predict the evolutionarily stable fraction of FDL (pESS). The model accounts for added winter mortality of SDL, general and kin competition among caterpillars, a competitive advantage of SDL over newly entering FDL (priority effect), and the avoidance of renewed infection of ant nests by butterflies in the coming season (segregation). We come to the following conclusions: (1) all factors listed above can promote the evolution of delayed development; (2) kin competition and segregation stabilize pESS near 0.5; and (3) a priority effect is the only mechanism potentially selecting for pESS < 0.5. However, given the empirical data, pESS is predicted to fall closer to 0.5 than to the 0.25 that has been observed. In this particular system, bet hedging cannot explain why more than 50% of larvae postpone growth. Presumably, other fitness benefits for SDL, for example, higher fertility or longevity, also contribute to the evolution of delayed development. The model presented here may be of general applicability for systems where maturing individuals compete in small subgroups.


Effect of short term graded withdrawal of nasal continuous positive airway pressure on systemic blood pressure in patients with obstructive sleep apnoea.
It is debated whether obstructive sleep apnoea (OSA) is a significant independent risk factor for sustained hypertension or cardiovascular morbidity and mortality. In an attempt to avoid the problem of confounding variables we have investigated whether withdrawing nasal continuous positive airway pressure (NCPAP) from patients with OSA for different proportions of the night leads to a subsequent rise in their morning blood pressures. Six patients with treated OSA had their NCPAP automatically varied between 3 cms H2O and a therapeutic pressure over 5 successive nights. The proportion of therapeutic NCPAP given was kept constant over the 5 nights and blood pressure measured the morning after the 5th night. Each patient had 5 different levels of sleep disruption, from no therapeutic NCPAP at all, through to 100% NCPAP. The nocturnal consequences of these different proportions of NCPAP were quantified both by oximetry and by a new EEG analysis that provides an objective estimate of the periodicity (fluctuations in the EEG depth) of the time course seen in patients with OSA. Increasing degrees of nocturnal hypoxic dipping and EEG periodicity were positively correlated with the subsequent morning systolic and diastolic blood pressures (p < 0.02). About 20% of the variance in systolic and diastolic blood pressure could be accounted for by the amount of either hypoxic dipping or EEG periodicity. The results of this study suggest that acute changes in awake blood pressure can be caused by sleep apnoea. It agrees with other data suggesting that OSA can have an independent influence on morning BP, but that this effect may have worn off by the afternoon and evening. Some of the discrepancies between the numerous studies in this area may be due to the timing of blood pressure measurements.


Bullies, gangs, drugs, and school: understanding the overlap and the role of ethnicity and urbanicity.
Recent media attention has increased interest in behavioral, mental health, and academic correlates of involvement in bullying. Yet, there has not been much interest in investigating the co-occurrence of other health-risk behaviors, such as gang membership, weapon carrying, and substance use. The potential influence of contextual factors, such as youth ethnicity, urbanicity, and school characteristics, also has been overlooked in previous research. The current study examined different subtypes of involvement in bullying-as primarily a victim, as primarily a bully, as both a victim and bully, and no involvement-and the association with significant health-risk behaviors, including engaging in violence and substance use, as well as academic problems. The analyses use self-report data from 16,302 adolescents (50.3 % female, 62.2 % Caucasian, 37.8 % African American) enrolled in 52 high schools. A series of three-level HLM analyses revealed that bullies and bully/victims were generally at greatest of risk of being involved in violence, engaging in multiple types of substance use, and having academic problems. These findings extend prior research by emphasizing a potential link between involvement in bullying and multiple health-risk behaviors, particularly among urban and African American high school youth.


Do medicines OSCEs improve drug administration ability?
To examine the efficacy of the early introduction of a medicines management 'objective structured clinical examination' (OSCE) into an undergraduate adult nursing students' programme of education and to determine if the acquisition of applied drug/pharmacology knowledge and drug administration of medicines had improved prior to qualification. A longitudinal comparative design was selected for this study. A convenience random samples method of three cohorts of adult nursing students (N=90) undertaking a pre-registration nursing programme was used to identify participants. Participants were assessed at the following points during their preparatory educational programmes. Cohort one (N=30) had completed 8 months of their programme, cohort two (N=30) had completed 20 months of their programme and cohort three (N=30) had completed 30 months of education. Students were asked to undertake a drug administration simulated activity (DASA) to generate the data for this study and differences between the students' performance was compared against the criteria of the DASA. There were statistical differences observed on almost every criteria of the DASA between the three groups of students. Students who had been previously exposed to medicines management OSCEs as an assessment method demonstrated superior medicines management skills and pharmacology knowledge than the control group (group three). The authors of this study conclude that early introduction of clinical examinations, namely OSCEs with an integrated approach to pharmacology and medicines management teaching, does facilitate and improve students drug administration and applied pharmacology ability.


Estimation of the monthly average daily solar radiation using geographic information system and advanced case-based reasoning.
The photovoltaic (PV) system is considered an unlimited source of clean energy, whose amount of electricity generation changes according to the monthly average daily solar radiation (MADSR). It is revealed that the MADSR distribution in South Korea has very diverse patterns due to the country's climatic and geographical characteristics. This study aimed to develop a MADSR estimation model for the location without the measured MADSR data, using an advanced case based reasoning (CBR) model, which is a hybrid methodology combining CBR with artificial neural network, multiregression analysis, and genetic algorithm. The average prediction accuracy of the advanced CBR model was very high at 95.69%, and the standard deviation of the prediction accuracy was 3.67%, showing a significant improvement in prediction accuracy and consistency. A case study was conducted to verify the proposed model. The proposed model could be useful for owner or construction manager in charge of determining whether or not to introduce the PV system and where to install it. Also, it would benefit contractors in a competitive bidding process to accurately estimate the electricity generation of the PV system in advance and to conduct an economic and environmental feasibility study from the life cycle perspective.


Estimating Catastrophic Costs due to Pulmonary Tuberculosis in Bangladesh.
To eliminate TB from the country by the year 2030, the Bangladesh National Tuberculosis (TB) Program is providing free treatment to the TB patients since 1993. However, the patients are still to make Out-of-their Pocket (OOP) payment, particularly before their enrollment Directly Observed Treatment Short-course (DOTS). This places a significant economic burden on poor-households. We, therefore, aimed to estimate the Catastrophic Health Expenditure (CHE) due to TB as well as understand associated difficulties faced by the families when a productive family member age (15-55) suffers from TB. The majority of the OOP expenditures occur before enrolling in. We conducted a cross-sectional study using multistage sampling in the areas of Bangladesh where Building Resources Across Communities (BRAC) provided TB treatment during June 2016. In total, 900 new TB patients, aged 15-55 years, were randomly selected from a list collected from BRAC program. CHE was defined as the OOP payments that exceeded 10% of total consumption expenditure of the family and 40% of total non-food expenditure/capacity-to-pay. Regular and Bayesian simulation techniques with 10,000 replications of re-sampling with replacement were used to examine robustness of the study findings. We also used linear regression and logit model to identify the drivers of OOP payments and CHE, respectively. The average total cost-of-illness per patient was 124 US$, of which 68% was indirect cost. The average CHE was 4.3% of the total consumption and 3.1% of non-food expenditure among the surveyed households. The poorest quintile of the households experienced higher CHE than their richest counterpart, 5% vs. 1%. Multiple regression model showed that the risk of CHE increased among male patients with smear-negative TB and delayed enrolling in the DOTS. Findings suggested that specific groups are more vulnerable to CHE who needs to be brought under innovative safety-net schemes.


Clostridium difficile contamination of public tap water distribution system during a waterborne outbreak in Finland.
In November through December 2007, the drinking water distribution system in the town of Nokia, Finland, was contaminated with treated sewage effluent that resulted in a large gastroenteritis outbreak in the community. The aim of the present study was to investigate if the contaminated water in this outbreak was also a potential source of Clostridium difficile infections. Samples from the contaminated tap water and treated sewage effluent were collected. Stool samples from a portion of patients that fell ill during the outbreak were examined for C. difficile. PCR ribotyping was performed on toxin positive C. difficile isolates and the genetic profiles of the water and patient isolates were compared. Twelve toxin-positive C. difficile isolates were found in water samples: five from contaminated tap water and seven from treated sewage effluent. Among these, four and five distinct PCR ribotype profiles were identified, respectively. Four PCR ribotype profiles were found among nine human faecal C. difficile isolates. Two isolates, one from tap water and one from a patient, had an indistinguishable PCR ribotype profile. Our findings demonstrate for the first time C. difficile contamination of a tap water distribution system and waterborne transmission of toxigenic C. difficile seems possible.


Effects of precursor topology and synthesis under crowding conditions on the structure of single-chain polymer nanoparticles.
By means of molecular dynamics simulations, we investigate the formation of single-chain nanoparticles through intramolecular cross-linking of polymer chains, in the presence of their precursors acting as purely steric crowders in concentrated solution. In the case of linear precursors, the structure of the resulting SCNPs is weakly affected by the density at which the synthesis is performed. Crowding has significant effects if ring precursors are used: higher concentrations lead to the formation of SCNPs with more compact and spherical morphologies. Such SCNPs retain in the swollen state (high dilution) the crumpled globular conformations adopted by the ring precursors in the crowded solutions. Increasing the concentration of both the linear and ring precursors up to 30% leads to faster formation of the respective SCNPs, prior to deceleration expected at higher densities. The results presented here propose promising new routes for the synthesis of globular SCNPs, which are usually elusive by conventional methods.


Chemical insights into bioinks for 3D printing.
3D printing has triggered the acceleration of numerous research areas in health sciences, which traditionally used cells as starting materials, in particular tissue engineering, regenerative medicine and also in the design of more relevant bioassays for drug discovery and development. While cells can be successfully printed in 2D layers without the help of any supporting biomaterial, the obtainment of more complex 3D architectures requires a specific bioink, i.e. a material in which the cells are embedded during and after the printing process helping to support them while they are arranged in superimposed layers. The bioink plays a critical role in bioprinting: first, it must be adapted to the 3D printing technology; then, it must fulfil the physicochemical and mechanical characteristics of the target construct (e.g. stiffness, elasticity, robustness, transparency); finally it should guarantee cell viability and eventually induce a desired behaviour. This review focuses on the nature of bioink components of natural or synthetic origin, and highlights the chemistry required for the establishment of the 3D network in conditions compatible with the selected 3D printing technique and cell survival.


Chronic Rhodiola rosea extract supplementation enforces exhaustive swimming tolerance.
We explored the effects and mechanisms of Rhodiola rosea extract supplementation on swimming-induced fatigue in rats. The concentrations of active components in Rhodiola rosea have been determined by high performance liquid chromatography-mass spectrometer. The Rhodiola rosea extract supplementation in water for 2-4 weeks was evaluated in male Wistar rats with 90-min unloaded swimming exercise and 5% body weight loaded swimming up to fatigue. We measured the fatigue biomarkers, including blood urea nitrogen (BUN), glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT), lactate dehydrogenase (LDH), hepatic glycogen content, the activity of fat metabolism enzymes, sterol regulatory element-binding protein-1 (SREBP-1) and fatty acid synthase (FAS), the tissue oxygen content and ratio of red and white skeletal muscle fibers in rats. Rhodiola rosea significantly increased liver glycogen, SREBP-1, FAS, heat shock protein 70 expression, Bcl-2/Bax ratio and oxygen content before swimming. Rhodiola rosea supplementation significantly increased the swimming time in a dose-dependent manner and reduced swimming-enhanced serum BUN, GOT and GPT levels. The ratio of red and white muscle fibers was not altered after chronic Rhodiola rosea extract supplementation. Chronic Rhodiola rosea supplementation significantly improved exhaustive swimming-induced fatigue by the increased glycogen content, energy supply of lipogenic enzyme expressions and protective defense mechanisms.


miRNA Profile in Three Different Normal Human Ocular Tissues by miRNA-Seq.
Because microRNAs (miRNAs) have been associated with eye diseases, our study aims to profile ocular miRNA expression in normal human ciliary body (CB), cornea, and trabecular meshwork (TM) using miRNA-Seq to provide a foundation for better understanding of miRNA function and disease involvement in these tissues. Total RNAs were extracted from seven normal human CB, seven cornea, and seven TM samples using mirVana total RNA isolation kit. miRNA-Seq was done with Illumina MiSeq. Bowtie software was used to trim and align generated sequence reads, and only exact matches to mature miRNAs from miRBase were included. The miRTarBase database was used to analyze miRNA target interactions, and the expression of five selected miRNAs was validated using droplet digital PCR (ddPCR). Using the miRNA extracted from 21 human samples, we found 378 miRNAs collectively expressed, of which the 11 most abundant miRNAs represented 80% of the total normalized reads. We also identified uniquely expressed miRNAs, of which five share 18 highly validated gene targets, and created a profile of miRNAs known to target genes associated with keratoconus and glaucoma. Using ddPCR, we validated the expression profile of five miRNAs from miRNA-Seq. For the first time, we profiled miRNA expression in three human ocular tissues using miRNA-Seq, identifying many miRNAs that had not been previously reported in ocular tissue. Defining the relative expression of miRNAs in nondiseased eye tissues could help uncover changes in miRNA expression that accompany diseases such as glaucoma and keratoconus.


Circuit Quantum Electrodynamics in Hyperbolic Space: From Photon Bound States to Frustrated Spin Models.
Circuit quantum electrodynamics is one of the most promising platforms for efficient quantum simulation and computation. In recent groundbreaking experiments, the immense flexibility of superconducting microwave resonators was utilized to realize hyperbolic lattices that emulate quantum physics in negatively curved space. Here we investigate experimentally feasible settings in which a few superconducting qubits are coupled to a bath of photons evolving on the hyperbolic lattice. We compare our numerical results for finite lattices with analytical results for continuous hyperbolic space on the Poincaré disk. We find good agreement between the two descriptions in the long-wavelength regime. We show that photon-qubit bound states have a curvature-limited size. We propose to use a qubit as a local probe of the hyperbolic bath, for example, by measuring the relaxation dynamics of the qubit. We find that, although the boundary effects strongly impact the photonic density of states, the spectral density is well described by the continuum theory. We show that interactions between qubits are mediated by photons propagating along geodesics. We demonstrate that the photonic bath can give rise to geometrically frustrated hyperbolic quantum spin models with finite-range or exponentially decaying interaction.


Structure of human cystathionine beta-synthase: a unique pyridoxal 5'-phosphate-dependent heme protein.
Cystathionine beta-synthase (CBS) is a unique heme- containing enzyme that catalyzes a pyridoxal 5'-phosphate (PLP)-dependent condensation of serine and homocysteine to give cystathionine. Deficiency of CBS leads to homocystinuria, an inherited disease of sulfur metabolism characterized by increased levels of the toxic metabolite homocysteine. Here we present the X-ray crystal structure of a truncated form of the enzyme. CBS shares the same fold with O-acetylserine sulfhydrylase but it contains an additional N-terminal heme binding site. This heme binding motif together with a spatially adjacent oxidoreductase active site motif could explain the regulation of its enzyme activity by redox changes.


A secondary kinetic isotope effect study of the 1-deoxy-D-xylulose-5-phosphate reductoisomerase-catalyzed reaction: evidence for a retroaldol-aldol rearrangement.
1-Deoxy-d-xylulose 5-phosphate (DXP) reductoisomerase (DXR, also known as methyl-d-erythritol 4-phosphate (MEP) synthase) is a NADPH-dependent enzyme, which catalyzes the conversion of DXP to MEP in the nonmevalonate pathway of isoprene biosynthesis. Two mechanisms have been proposed for the DXR-catalyzed reaction. In the alpha-ketol rearrangement mechanism, the reaction begins with deprotonation of the C-3 hydroxyl group followed by a 1,2-migration to give methylerythrose phosphate, which is then reduced to MEP by NADPH. In the retroaldol/aldol rearrangement mechanism, DXR first cleaves the C3-C4 bond of DXP in a retroaldol manner to generate a three-carbon and a two-carbon phosphate bimolecular intermediate. These two species are then reunited by an aldol reaction to form a new C-C bond, yielding an aldehyde intermediate. Subsequent reduction by NADPH affords MEP. To differentiate these mechanisms, we have prepared [3-(2)H]- and [4-(2)H]-DXP and carried out a competitive secondary kinetic isotope effect (KIE) study of the DXR reaction. The normal 2 degrees KIEs observed for [3-(2)H]- and [4-(2)H]-DXP provide compelling evidence supporting a retroaldol/aldol mechanism for the rearrangement catalyzed by DXR, with the rate-limiting step being cleavage of the C3-C4 bond of DXP.


A strategy for the comparative analysis of serum proteomes for the discovery of biomarkers for hepatocellular carcinoma.
Many of the emerging technologies for the global evaluation of gene expression, at both the RNA and protein level, are being applied to the problem of finding biomarkers for human disease progression. These analyses can be made difficult, however, by variation between samples that arises from both technical and nondisease related physiological or genetic causes. In an effort to identify serum polypeptides whose presence or absence correlates with the clinical status of patients at high risk for hepatocellular carcinoma (HCC), we have developed a strategy that helps to focus the analysis on meaningful changes in protein levels above the background of variation. For the current study we divided the patient population into four clinically defined diagnostic groups that represent a generally increasing risk for HCC. Chronic infection with hepatitis B virus (HBV) is a major risk factor for HCC and our groups included patients with no indication of liver disease (healthy), those with inactive chronic HBV, those with active chronic HBV, and patients with a diagnosis of HCC and history of chronic HBV infection. Serum polypeptides from these patients were first analyzed in two-dimensional gels by combining the serum from patients in each of the four groups to generate composite gel profiles. Analysis of these composite gels allowed us to identify two relatively abundant features that were reduced in the HCC group as compared to the healthy group. Tryptic fragment mass fingerprinting identified the features as a carboxy terminal fragment of complement C3 and an isoform of apolipoprotein A1. These two features were examined by two-dimensional gel electrophoresis of serum from each individual in the four groups in order to verify that the inter-group differences seen in composite gels reported changes in abundance for most members of the group, rather than extreme changes for a small fraction of the group. These preliminary studies suggest that a proteomic methodology can be used for the identification of serum biomarkers for HCC and other liver disease.


Angiotensin II is mitogenic in neonatal rat cardiac fibroblasts.
Angiotensin II has been reported to be a hormonal stimulus of cardiac growth, a response that may involve myocyte hypertrophy as well as growth of nonmyocytes. This study was designed to determine whether neonatal rat cardiac fibroblasts have an angiotensin II receptor that is coupled with hypertrophic and/or proliferative growth. Competitive radioligand binding studies showed that cardiac fibroblasts have a single class of high-affinity (IC50, 1.0 nM) angiotensin II binding sites (Bmax, 778 fmol/mg protein) that are sensitive to the competitive nonpeptide AT1 receptor antagonist losartan (IC50, 13 nM). Other angiotensin peptides competed for [125I]angiotensin II binding in the following rank order: angiotensin II > angiotensin III > angiotensin I > > [des-Asp1-des-Arg2]angiotensin II. A nonhydrolyzable analogue of guanosine triphosphate increased the dissociation rate of bound [125I]angiotensin II and decreased hormone binding to the receptor at equilibrium. The angiotensin II receptor was coupled with increases in intracellular calcium. Incorporation of precursors into protein, DNA, and RNA in response to angiotensin II was determined. In serum-deprived cultures, a 24-hour exposure to 1 microM [Sar1]angiotensin II increased rates of phenylalanine, thymidine, and uridine incorporation by 58%, 103%, and 118%, respectively. These increases were blocked by the noncompetitive AT1 receptor antagonist EXP3174. After 48 hours, [Sar1]angiotensin II increased total protein and DNA of cardiac fibroblasts by 23% and 15%, respectively, with no change in the protein/DNA ratio. [Sar1]Angiotensin II increased cell number by 138% after a 24-hour exposure, without affecting cell area. In summary, cardiac fibroblasts have G protein-linked AT1 receptors that are coupled with proliferative growth. These results suggest that angiotensin II-induced cardiac hypertrophy is, in part, secondary to stimulated increases in nonmyocyte cellular growth.


Adaptive plasticity in wild field cricket's acoustic signaling.
Phenotypic plasticity can be adaptive when phenotypes are closely matched to changes in the environment. In crickets, rhythmic fluctuations in the biotic and abiotic environment regularly result in diel rhythms in density of sexually active individuals. Given that density strongly influences the intensity of sexual selection, we asked whether crickets exhibit plasticity in signaling behavior that aligns with these rhythmic fluctuations in the socio-sexual environment. We quantified the acoustic mate signaling behavior of wild-caught males of two cricket species, Gryllus veletis and G. pennsylvanicus. Crickets exhibited phenotypically plastic mate signaling behavior, with most males signaling more often and more attractively during the times of day when mating activity is highest in the wild. Most male G. pennsylvanicus chirped more often and louder, with shorter interpulse durations, pulse periods, chirp durations, and interchirp durations, and at slightly higher carrier frequencies during the time of the day that mating activity is highest in the wild. Similarly, most male G. veletis chirped more often, with more pulses per chirp, longer interpulse durations, pulse periods, and chirp durations, shorter interchirp durations, and at lower carrier frequencies during the time of peak mating activity in the wild. Among-male variation in signaling plasticity was high, with some males signaling in an apparently maladaptive manner. Body size explained some of the among-male variation in G. pennsylvanicus plasticity but not G. veletis plasticity. Overall, our findings suggest that crickets exhibit phenotypically plastic mate attraction signals that closely match the fluctuating socio-sexual context they experience.


Mucosal Immune Responses to Respiratory Syncytial Virus.
Despite over half a century of research, respiratory syncytial virus (RSV)-induced bronchiolitis remains a major cause of hospitalisation in infancy, while vaccines and specific therapies still await development. Our understanding of mucosal immune responses to RSV continues to evolve, but recent studies again highlight the role of Type-2 immune responses in RSV disease and hint at the possibility that it dampens Type-1 antiviral immunity. Other immunoregulatory pathways implicated in RSV disease highlight the importance of focussing on localised mucosal responses in the respiratory mucosa, as befits a virus that is essentially confined to the ciliated respiratory epithelium. In this review, we discuss studies of mucosal immune cell infiltration and production of inflammatory mediators in RSV bronchiolitis and relate these studies to observations from peripheral blood. We also discuss the advantages and limitations of studying the nasal mucosa in a disease that is most severe in the lower airway. A fresh focus on studies of RSV pathogenesis in the airway mucosa is set to revolutionise our understanding of this common and important infection.


Time Series Analysis of the Microbiota of Children Suffering From Acute Infectious Diarrhea and Their Recovery After Treatment.
Gut microbiota is closely related to acute infectious diarrhea, one of the leading causes of mortality and morbidity in children worldwide. Understanding the dynamics of the recovery from this disease is of clinical interest. This work aims to correlate the dynamics of gut microbiota with the evolution of children who were suffering from acute infectious diarrhea caused by a rotavirus, and their recovery after the administration of a probiotic, Saccharomyces boulardii CNCM I-745. The experiment involved 10 children with acute infectious diarrhea caused by a rotavirus, and six healthy children, all aged between 3 and 4 years. The children who suffered the rotavirus infection received S. boulardii CNCM I-745 twice daily for the first 5 days of the experiment. Fecal samples were collected from each participant at 0, 3, 5, 10, and 30 days after probiotic administration. Microbial composition was characterized by 16S rRNA gene sequencing. Alpha and beta diversity were calculated, along with dynamical analysis based on Taylor's law to assess the temporal stability of the microbiota. All children infected with the rotavirus stopped having diarrhea at day 3 after the intervention. We observed low alpha diversities in the first 5 days (p-value < 0.05, Wilcoxon test), larger at 10 and 30 days after probiotic treatment. Canonical correspondence analysis (CCA) showed differences in the gut microbiota of healthy children and of those who suffered from acute diarrhea in the first days (p-value < 0.05, ADONIS test), but not in the last days of the experiment. Temporal variability was larger in children infected with the rotavirus than in healthy ones. In particular, Gammaproteobacteria class was found to be abundant in children with acute diarrhea. We identified the microbiota transition from a diseased state to a healthy one with time, whose characterization may lead to relevant clinical data. This work highlights the importance of using time series for the study of dysbiosis related to diarrhea.


Diagnosis and kidney-sparing treatments for upper tract urothelial carcinoma: state of the art.
Conservative management of upper tract urothelial cancer (UTUC) is becoming increasingly popular: the key to success is correct selection of patients with low-risk UTUC based on size (≤2 cm), focality (single lesion), stage (< T2), and grade (low grade). Despite the recent growing interest in the conservative approach to UTUC, the diagnostic process is still a challenge, and kidney-sparing surgery (KSS) is traditionally reserved for patients with contraindications to radical nephroureterectomy. In order to explore the "state of the art" in the diagnosis and conservative treatment of UTUC, a systematic review of the literature was performed. A PubMed, Scopus, and Cochrane search for peer-reviewed studies was performed using the keywords "upper tract urothelial carcinoma" OR "UTUC" OR "upper urinary tract" AND "biopsy" OR "diagnosis" OR "endomicroscopy" OR "imaging" AND "URS" OR "ureteroscopy" OR "kidney-sparing surgery" OR "laser ablation" OR "ureterectomy". We considered as relevant comparative prospective studies (randomized, quasi-randomized, no randomized), retrospective studies, meta-analyses, systematic reviews, and case report series written in the English language. Letters to the editor and contributions written in languages other than English were not considered of value for this review. Eligible articles were reviewed according to the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) criteria. Two hundred and sixty-three (263) records were identified using the above-mentioned keywords. Overall, 30 studies were considered relevant for the purpose of this systematic review and for the evidence evaluation process during qualitative synthesis. The outcomes evaluated in this review were the current diagnostic methods and the KSS approaches in UTUC. Furthermore, we included in the review the emerging technology for distinguishing between normal tissue, low-grade UTUC, and high-grade UTUC. Conclusive diagnosis is fundamental to the decision-making process in patients who could benefit from conservative treatment of UTUC. The most relevant diagnostic modalities are computed tomography urography, local urine cytology, and ureteroscopy with acquisition of an adequate biopsy sample for histology. KSS includes the endourological approach and segmental ureterectomy. Promising technology in the endourological management of UTUC helps in providing intraoperative information on UTUC grading and staging, with a high accuracy. Patients treated conservatively have to undergo stringent postoperative follow-up in order to detect and, if necessary, treat any recurrence promptly. Further larger and multicenter studies are needed to confirm these findings.


Developmental self-construction and -configuration of functional neocortical neuronal networks.
The prenatal development of neural circuits must provide sufficient configuration to support at least a set of core postnatal behaviors. Although knowledge of various genetic and cellular aspects of development is accumulating rapidly, there is less systematic understanding of how these various processes play together in order to construct such functional networks. Here we make some steps toward such understanding by demonstrating through detailed simulations how a competitive co-operative ('winner-take-all', WTA) network architecture can arise by development from a single precursor cell. This precursor is granted a simplified gene regulatory network that directs cell mitosis, differentiation, migration, neurite outgrowth and synaptogenesis. Once initial axonal connection patterns are established, their synaptic weights undergo homeostatic unsupervised learning that is shaped by wave-like input patterns. We demonstrate how this autonomous genetically directed developmental sequence can give rise to self-calibrated WTA networks, and compare our simulation results with biological data.


Tumor regression induced by intratumoral injection of DNA coding for human interleukin 12 into melanoma metastases in gray horses.
Preclinical studies investigating new therapeutic principles against melanoma are presently being carried out in mouse models; however, these are not optimal. Here we describe a novel animal model using gray horses. These animals spontaneously develop metastatic melanoma that resembles human disease and is thus highly relevant for preclinical studies testing new immunotherapy protocols. We found that injection of plasmid DNA coding for the human cytokine interleukin 12 into established metastases induced significant regression in all 12 treated lesions in a total of 7 horses. Complete disappearance was observed in one treated lesion, with no recurrence after 6 months. No adverse events have been observed in any of the animals during and after treatment. These results demonstrate the effectiveness and safety of interleukin 12 encoding plasmid DNA therapy against established metastatic disease in a large animal model and serve as a basis for a clinical trial.


The tolerance and efficacy of interferon-alpha in haemodialysis patients with HCV infection: a multicentre, prospective study.
A prospective multicentre study was initiated in HCV-infected haemodialysis patients to assess the tolerance and efficacy of alpha-2b interferon. We had planned to include 120 patients with HCV RNA detectable by polymerase chain reaction (PCR) (Amplicor Roche) and histologically documented chronic hepatitis. The dose of alpha-interferon was 3 million units (MU) three times weekly (TTW), to be reduced to 1.5 MU TTW in case of side-effects. Tolerance was evaluated monthly; virological efficacy was evaluated by PCR. A liver biopsy was performed at month 18 (M18). (a) After 37 patients had been included, the study was discontinued by the promoting institution because of severe side-effects requiring that treatment be stopped in 19 patients. The side-effects were: cardiac (4) neuropsychiatric (2), digestive (3), acute necrosis of the graft (1), severe asthenia (9), minor side-effects were observed in 22 patients. A complete 12-month course was completed in 12 patients for the 3 MU TTW dose and in six patients for the 1.5 MU TTW reduced dose. Normal ALT level (OR, 0.16; CI 95%, 0.03-0.89) at inclusion was associated with interruption of treatment (univariate analysis). (b) Sustained virological response was observed in only seven (18.9%), of the 18 patients who completed the treatment (38%). Increased ALT at inclusion (OR, 1.04; CI 95%, 1.01-1.09) and cumulated doses of interferon (OR, 1.01; CI 95%, 1.004-1.026) were jointly associated with a sustained response, while positive PCR at M2 was strongly predictive of treatment failure. Tolerance of interferon is poor in haemodialysis patients. Sustained response is fairly high in patients who have 12 months of treatment and seems to be based on the immune status of the patients (ALT) and the cumulative doses of interferon.


Primary carcinoma of the gallbladder.
The clinical and pathological features of 100 consecutive cases of primary carcinoma of the gallbladder, treated by a single surgical unit, are described. Females out numbered males by 2 to 1. The maximum incidence was seen in the fifth decade. Pain and the presence of a mass were the two most common clinical features. Gallstones coexisted in 45 per cent of cases. Ancillary investigations were of little value in preoperative diagnosis. At laparotomy biopsy alone was possible in a large number of cases owing to the advanced disease process. Two patients were alive 5 years after surgery. Prophylactic cholecystectomy for all diseased gallbladders, whether symptomatic or not, is advocated.


Postoperative Analgesic Efficacy of Preemptive and Postoperative Lornoxicam or Tramadol in Lumbar Disc Surgery.
To compare preemptive and postoperative analgesic efficacy of tramadol and lornoxicam administered before anaesthesia induction in lumbar discectomy. This randomised, double-blind trial was conducted on 60 ASA I and II patients undergoing lumbar discectomy. Group L (n=30) received 3×8 mg day-1 lornoxicam, and Group T (n=30) received 3×1.5 mg kg-1 day-1 tramadol. A verbal rating scale (VRS), the duration of effective analgesia, the number of additional analgesics used, adverse effects and patient satisfaction were evaluated at the postoperative 30th minute and 1st, 2nd, 4th, 6th, 8th, 12th and 24th hours. There were no significant differences between Groups L and T regarding demographic and clinical characteristics, the number of additional analgesics and the duration of effective analgesia, adverse effects and patient satisfaction. VRS scores of the patients in Group T were significantly higher than those in Group L at the postoperative 30th minute (p=0.050) and the 1st hour (p=0.005). Lornoxicam, which was used for preemptive and postoperative analgesia in lumbar disc surgery, had provided adequate and effective analgesia such as tramadol. Moreover, preemptive analgesia was quite effective in prevention and treatment of postoperative pain.


Pro-angiogenic capacities of microvesicles produced by skin wound myofibroblasts.
Wound healing is a very highly organized process where numerous cell types are tightly regulated to restore injured tissue. Myofibroblasts are cells that produce new extracellular matrix and contract wound edges. We previously reported that the human myofibroblasts isolated from normal wound (WMyos) produced microvesicles (MVs) in the presence of the serum. In this study, MVs were further characterized using a proteomic strategy and potential functions of the MVs were determined. MV proteins isolated from six WMyo populations were separated using two-dimensional differential gel electrophoresis. Highly conserved spots were selected and analyzed using mass spectrometry resulting in the identification of 381 different human proteins. Using the DAVID database, clusters of proteins involved in cell motion, apoptosis and adhesion, but also in extracellular matrix production (21 proteins, enrichment score: 3.32) and in blood vessel development/angiogenesis (19 proteins, enrichment score: 2.66) were identified. Another analysis using the functional enrichment analysis tool FunRich was consistent with these results. While the action of the myofibroblasts on extracellular matrix formation is well known, their angiogenic potential is less studied. To further characterize the angiogenic activity of the MVs, they were added to cultured microvascular endothelial cells to evaluate their influence on cell growth and migration using scratch test and capillary-like structure formation in Matrigel®. The addition of a MV-enriched preparation significantly increased endothelial cell growth, migration and capillary formation compared with controls. The release of microvesicles by the wound myofibroblasts brings new perspectives to the field of communication between cells during the normal healing process.


Pathways to ischemic neuronal cell death: are sex differences relevant?
We have known for some time that the epidemiology of human stroke is sexually dimorphic until late in life, well beyond the years of reproductive senescence and menopause. Now, a new concept is emerging: the mechanisms and outcome of cerebral ischemic injury are influenced strongly by biological sex as well as the availability of sex steroids to the brain. The principal mammalian estrogen (17 beta estradiol or E2) is neuroprotective in many types of brain injury and has been the major focus of investigation over the past several decades. However, it is becoming increasingly clear that although hormones are a major contributor to sex-specific outcomes, they do not fully account for sex-specific responses to cerebral ischemia. The purpose of this review is to highlight recent studies in cell culture and animal models that suggest that genetic sex determines experimental stroke outcome and that divergent cell death pathways are activated after an ischemic insult. These sex differences need to be identified if we are to develop efficacious neuroprotective agents for use in stroke patients.


Polymorphisms of toll-like receptors and their pathways in viral hepatitis.
Toll-like receptors (TLRs) are an important part of the innate immune response to a variety of pathogens including hepatic viral infections. Activation of TLRs stimulates a complex intracellular signalling cascade that results in production of proinflammatory cytokines and interferons important for antiviral responses as well as induction of the adaptive arm of the immune system. There is substantial evidence for an important role for TLRs and TLR-mediated signalling in the pathogenesis and outcomes of hepatitis B and C in particular, but it might also influence responses to other viral hepatitis infections. Several single nucleotide polymorphisms (SNPs) of TLRs, relevant adaptor molecules and cytokines mediated by TLR signalling have been described that alter innate immune responses and have been implicated in a variety of human diseases including viral and other infections. There is now significant evidence that a number of TLR SNPs can affect various clinical outcomes in Caucasian patients with chronic HCV. However, the role of these polymorphisms in acute and other chronic hepatitis infections, including HBV as well as in non-Caucasian populations, has not been elucidated. In addition, results for SNPs downstream of TLR activation, such as in relevant cytokines, are inconsistent and their influence requires further investigation to determine the clinical significance of genetic variations in these mediators.


Fatty liver disease, an emerging etiology of hepatocellular carcinoma in Argentina.
To investigate any changing trends in the etiologies of hepatocellular carcinoma (HCC) in Argentina during the last years. A longitudinal cohort study was conducted by 14 regional hospitals starting in 2009 through 2016. All adult patients with newly diagnosed HCC either with pathology or imaging criteria were included. Patients were classified as presenting non-alcoholic fatty liver disease (NAFLD) either by histology or clinically, provided that all other etiologies of liver disease were ruled out, fatty liver was present on abdominal ultrasound and alcohol consumption was excluded. Complete follow-up was assessed in all included subjects since the date of HCC diagnosis until death or last medical visit. A total of 708 consecutive adults with HCC were included. Six out of 14 hospitals were liver transplant centers (n = 484). The prevalence of diabetes mellitus was 27.7%. Overall, HCV was the main cause of liver disease related with HCC (37%) including cirrhotic and non-cirrhotic patients, followed by alcoholic liver disease 20.8%, NAFLD 11.4%, cryptogenic 9.6%, HBV 5.4% infection, cholestatic disease and autoimmune hepatitis 2.2%, and other causes 9.9%. A 6-fold increase in the percentage corresponding to NAFLD-HCC was detected when the starting year, i.e., 2009 was compared to the last one, i.e., 2015 (4.3% vs 25.6%; P < 0.0001). Accordingly, a higher prevalence of diabetes mellitus was present in NAFLD-HCC group 61.7% when compared to other than NAFLD-HCC 23.3% (P < 0.0001). Lower median AFP values at HCC diagnosis were observed between NAFLD-HCC and non-NAFLD groups (6.6 ng/mL vs 26 ng/mL; P = 0.02). Neither NAFLD nor other HCC etiologies were associated with higher mortality. The growing incidence of NAFLD-HCC documented in the United States and Europe is also observed in Argentina, a confirmation with important Public Health implications.


Reading Instructions Influence Cognitive Processes of Illustrated Text Reading Not Subject Perception: An Eye-Tracking Study.
Scientific illustrations play an important role in scientific texts, however, young readers show limited ability to use illustration information and integrate it with the text in multimedia learning. The primary goal of the present study was to investigate if strategy instructions for illustrated text reading focused on scientific illustration reading and text-illustration integration can help young readers overcome their deficiencies and change their reading processes, learning outcomes, and subjective perceptions of article difficulty and enjoyment, illustration enjoyment, and self-evaluation of learning. Besides, is subjective perception of the article related to reading behavior? Sixty-two fourth-grade students read an illustrated science text while their eye movements were recorded, and then completed a reading test and questionnaire. The instruction group outperformed the control group on the reading test, but subjective perceptions of the article did not differ between groups. Eye-movements analysis showed that the instruction group spent twice as much time reading the illustrations and made more saccades between relevant text and illustration sections than the control group. These findings indicate that strategy instructions for reading illustrated text promoted reading comprehension and changed learning processes, not subjective perceptions. In addition, the result of this study showed that there was no relationship between subjective perception (article difficulty or illustrations enjoyment) of the article and reading time (total reading time on the illustrated science text and on the science illustrations). This study had empirical and practical contributions. Empirically, this study developed the instruction methods in multimedia learning and further examined their effect on learning processes in young readers. Practically, this study can help elementary school teachers understand the processes used by young readers when reading illustrated texts and provide them with evidence-based instructions to teach science reading effectively.


Monitoring the penetration process of single microneedles with varying tip diameters.
Microneedles represent promising tools for delivery of drugs to the skin. However, before these microneedles can be used in clinical practice, it is essential to understand the process of skin penetration by these microneedles. The present study was designed to monitor both penetration depth and force of single solid microneedles with various tip diameters ranging from 5 to 37µm to provide insight into the penetration process into the skin of these sharp microneedles. To determine the microneedle penetration depth, single microneedles were inserted in human ex vivo skin while monitoring the surface of the skin. Simultaneously, the force on the microneedles was measured. The average penetration depth at 1.5mm displacement was similar for all tip diameters. However, the process of penetration depth was significantly different for the various microneedles. Microneedles with a tip diameter of 5µm were smoothly inserted into the skin, while the penetration depth of microneedles with a larger tip diameter suddenly increased after initial superficial penetration. In addition, the force at insertion (defined as the force at a sudden decrease in measured force) linearly increased with tip diameter ranging from 20 to 167mN. The force drop at insertion was associated with a measured penetration depth of approximately 160μm for all tip diameters, suggesting that the drop in force was due to the penetration of a deeper skin layer. This study showed that sharp microneedles are essential to insert microneedles in a well-controlled way to a desired depth.


Assembly reflects evolution of protein complexes.
A homomer is formed by self-interacting copies of a protein unit. This is functionally important, as in allostery, and structurally crucial because mis-assembly of homomers is implicated in disease. Homomers are widespread, with 50-70% of proteins with a known quaternary state assembling into such structures. Despite their prevalence, their role in the evolution of cellular machinery and the potential for their use in the design of new molecular machines, little is known about the mechanisms that drive formation of homomers at the level of evolution and assembly in the cell. Here we present an analysis of over 5,000 unique atomic structures and show that the quaternary structure of homomers is conserved in over 70% of protein pairs sharing as little as 30% sequence identity. Where quaternary structure is not conserved among the members of a protein family, a detailed investigation revealed well-defined evolutionary pathways by which proteins transit between different quaternary structure types. Furthermore, we show by perturbing subunit interfaces within complexes and by mass spectrometry analysis, that the (dis)assembly pathway mimics the evolutionary pathway. These data represent a molecular analogy to Haeckel's evolutionary paradigm of embryonic development, where an intermediate in the assembly of a complex represents a form that appeared in its own evolutionary history. Our model of self-assembly allows reliable prediction of evolution and assembly of a complex solely from its crystal structure.


Frailty Index Developed From a Cancer-Specific Geriatric Assessment and the Association With Mortality Among Older Adults With Cancer.
Background: An objective measure is needed to identify frail older adults with cancer who are at increased risk for poor health outcomes. The primary objective of this study was to develop a frailty index from a cancer-specific geriatric assessment (GA) and evaluate its ability to predict all-cause mortality among older adults with cancer. Patients and Methods: Using a unique and novel data set that brings together GA data with cancer-specific and long-term mortality data, we developed the Carolina Frailty Index (CFI) from a cancer-specific GA based on the principles of deficit accumulation. CFI scores (range, 0-1) were categorized as robust (0-0.2), pre-frail (0.2-0.35), and frail (>0.35). The primary outcome for evaluating predictive validity was all-cause mortality. The Kaplan-Meier method and log-rank tests were used to compare survival between frailty groups, and Cox proportional hazards regression models were used to evaluate associations. Results: In our sample of 546 older adults with cancer, the median age was 72 years, 72% were women, 85% were white, and 47% had a breast cancer diagnosis. Overall, 58% of patients were robust, 24% were pre-frail, and 18% were frail. The estimated 5-year survival rate was 72% in robust patients, 58% in pre-frail patients, and 34% in frail patients (log-rank test, P<.0001). Frail patients had more than a 2-fold increased risk of all-cause mortality compared with robust patients (adjusted hazard ratio, 2.36; 95% CI, 1.51-3.68). Conclusions: The CFI was predictive of all-cause mortality in older adults with cancer, a finding that was independent of age, sex, cancer type and stage, and number of medical comorbidities. The CFI has the potential to become a tool that oncologists can use to objectively identify frailty in older adults with cancer.


Ligand modulated antagonism of PPARgamma by genomic and non-genomic actions of PPARdelta.
Members of the Peroxisome Proliferator Activated Receptor, PPAR, subfamily of nuclear receptors display complex opposing and overlapping functions and a wide range of pharmacological and molecular genetic tools have been used to dissect their specific functions. Non-agonist bound PPARdelta has been shown to repress PPAR Response Element, PPRE, signalling and several lines of evidence point to the importance of PPARdelta repressive actions in both cardiovascular and cancer biology. In this report we have employed transient transfections and luciferase reporter gene technology to study the repressing effects of PPARdelta and two derivatives thereof. We demonstrate for the first time that the classical dominant negative deletion of the Activation Function 2, AF2, domain of PPARdelta show enhanced repression of PPRE signalling in the presence of a PPARdelta agonist. We propose that the mechanism for the phenomenon is increased RXR heterodimerisation and DNA binding upon ligand binding concomitant with transcriptional co-repressor binding. We also demonstrated ligand-dependent dominant negative action of a DNA non-binding derivative of PPARdelta on PPARgamma1 signalling. This activity was abolished upon over-expression of RXRalpha suggesting a role for PPAR/cofactor competition in the absence of DNA binding. These findings are important in understanding the wide spectrum of molecular interactions in which PPARdelta and PPARgamma have opposing biological roles and suggest novel paradigms for the design of different functional classes of nuclear receptor antagonist drugs.


Directional edge enhancement in optical tomography of thin phase objects.
In this paper, we make a proposal to obtain the Hilbert-transform for each entry of the projection data leaving the slice of a thin phase object. These modified projections are stacked in such a way that they form a modified sinogram called Hilbert-sinogram. We prove that the inverse Radon-transform of this sinogram is the directional Hilbert-transform of the slice function, and the reconstructed image is the directional edge enhancement of the distribution function on the slice. The Hilbert-transform is implemented by a 4f optical Fourier-transform correlator and a spatial filter consisting of a phase step of π radians. One important feature of this proposal is to perform a turn of 180° in the spatial filter at a certain value of the projection angle within the range [0°, 360°]. The desired direction of enhancement can be chosen by the proper selection of such turning angle. We present both the mathematical modeling and numerical results.