Uncoupling protein-2 mRNA expression in mice subjected to intermittent hypoxia.
To investigate the effect of intermittent hypoxia-a model of obstructive sleep apnea (OSA)-on pancreatic expression of uncoupling protein-2 (UCP2), as well as on glycemic and lipid profiles, in C57BL mice. For 8 h/day over a 35-day period, male C57BL mice were exposed to intermittent hypoxia (hypoxia group) or to a sham procedure (normoxia group). The intermittent hypoxia condition involved exposing mice to an atmosphere of 92% N and 8% CO2 for 30 s, progressively reducing the fraction of inspired oxygen to 8 ± 1%, after which they were exposed to room air for 30 s and the cycle was repeated (480 cycles over the 8-h experimental period). Pancreases were dissected to isolate the islets. Real-time PCR was performed with TaqMan assays. Expression of UCP2 mRNA in pancreatic islets was 20% higher in the normoxia group than in the hypoxia group (p = 0.11). Fasting serum insulin was higher in the hypoxia group than in the normoxia group (p = 0.01). The homeostasis model assessment of insulin resistance indicated that, in comparison with the control mice, the mice exposed to intermittent hypoxia showed 15% lower insulin resistance (p = 0.09) and 21% higher pancreatic β-cell function (p = 0.01). Immunohistochemical staining of the islets showed no significant differences between the two groups in terms of the area or intensity of α- and β-cell staining for insulin and glucagon. To our knowledge, this is the first report of the effect of intermittent hypoxia on UCP2 expression. Our findings suggest that UCP2 regulates insulin production in OSA. Further study of the role that UCP2 plays in the glycemic control of OSA patients is warranted.


The fast discrete Radon transform. I. Theory.
An inversion scheme for reconstruction of images from projections based on the slope-intercept form of the discrete Radon transform is presented. A seminal algorithm for the forward and the inverse transforms proposed by G. Beylkin (1987) demonstrated poor dispersion characteristics for steep slopes and could not invert transforms based on nonlinear slope variations. By formulating the computation of a discrete computation of the continuous Radon transform formula, the authors explicitly derive fast generalized inversion methods that overcome the original shortcomings. The generalized forward (FRT) and inverse (IFRT) algorithms proposed are fast, eliminate interpolation calculations, and convert directly between a raster scan grid and a rectangular/polar grid in one step.


A transient CRISPR/Cas9 expression system for genome editing in Trypanosoma brucei.
Generation of knockouts and in situ tagging of genes in Trypanosoma brucei has been greatly facilitated by using CRISPR/Cas9 as a genome editing tool. To date, this has entailed using a limited number of cell lines that are stably transformed to express Cas9 and T7 RNA polymerase (T7RNAP). It would be desirable, however, to be able to use CRISPR/Cas9 for any trypanosome cell line. We describe a sequential transfection expression system that enables transient expression of the two proteins, followed by delivery of PCR products for gRNAs and repair templates. This procedure can be used for genome editing without the need for stable integration of the Cas9 and T7RNAP genes.


[A modified method of nuclear transfer for investigating early development of mouse embryos reconstructed with cumulus cell nuclei].
To establish a fast, simple, efficient and minimally invasive method for nuclear transfer (NT) to study the early development of mouse embryos reconstructed with cumulus cell nuclei in vitro. With a sharp-tipped enucleation needle, an incision approximately 25 mm in length was made in the zona pellucida of a rat oocyte, through which the first polar body was removed and the metaphase II chromosome-spindle complex was gently aspirated along with a minimal volume of the cytoplasm by slightly pressing and vacuum aspiration of the oocyte. A cumulus cell nuclei from C57BL/6j mouse, 10-12 mm in diameter, was inserted into the perivitelline space of the enucleated oocyte. The fusion of the donor-recipient pair was induced by electrofusion and nuclear formation was observed. The development of 2-cell, 4- to 8-cell and morula-stage embryos was observed after a 72-hour culture of the reconstructed oocytes in vitro. The modified NT method enabled one-step removal of the whole nucleus from the oocyte with confirmed reliability of complete nuclear removal by Hoechst 33342 staining of the removed nuclei examined under UV light. The process of enucleation took an average time of 15 s, and the survival rate of the enucleated oocytes reached 95%. The success rate of 76.7% was achieved for cumulus cell nucleus insertion into the zona pellucida of the enucleated oocytes and pronucleus formation occurred in 62.2% of the reconstructed oocytes with nuclear transfer. After 72 h of culture in vitro of the reconstructed oocytes in CZB medium, the percentage of embryos that developed into 2-cell, 4- to 8-cell and morula (more than 16 cells) stages were 57.5%, 39.1% and 27.6%, respectively. Microsatellite sequences (D7Mit22 and D4Mit204) were amplified from the DNA of the reconstructed embryos for identifying their origin, which was proved to be C57BL/6j mouse. The modified NT method is simple, minimally invasive, efficient and practicable to reconstruct mouse embryos with somatic cell nuclei.


Circadian rhythms from multiple oscillators: lessons from diverse organisms.
The organization of biological activities into daily cycles is universal in organisms as diverse as cyanobacteria, fungi, algae, plants, flies, birds and man. Comparisons of circadian clocks in unicellular and multicellular organisms using molecular genetics and genomics have provided new insights into the mechanisms and complexity of clock systems. Whereas unicellular organisms require stand-alone clocks that can generate 24-hour rhythms for diverse processes, organisms with differentiated tissues can partition clock function to generate and coordinate different rhythms. In both cases, the temporal coordination of a multi-oscillator system is essential for producing robust circadian rhythms of gene expression and biological activity.


Determination of stability constants of U(VI)-Fe(III)-citrate complexes.
Ion-exchange experiments were performed to evaluate the formation of the uranium-citrate and uranium-iron-citrate complexes over a wide concentration range; i.e., environmentally relevant concentrations (e.g., 10(-6) M in metal and ligand) and concentrations useful for spectroscopic investigations (e.g., 10(-4) M in metal and ligand). The stability of the well-known uranium-citrate complex was determined to validate the computational and experimental methods applied to the more complex system. Values of the conditional stability constants for these species were obtained using a chemical equilibrium model in FITEQL. At a pH of 4.0, the stability constant for uranium-citrate complex (log beta1,1) was determined to be 8.71+/-0.6 at I = 0. Analysis of the results of ion-exchange experiments for the U-Fe-citric acid system indicates the formation of the 1:1:1 and 1:1:2 ternary species with stability constants (log beta) of 17.10+/-0.41 and 20.47+/-0.31, respectively, at I= 0.


Current Insights on the Treatment of Anaplastic Lymphoma Kinase-Positive Metastatic Non-Small Cell Lung Cancer: Focus on Brigatinib.
Rearrangement of anaplastic lymphoma kinase (ALK) gene is detected in approximately 5% of non-small cell lung cancer (NSCLC) patients. Tyrosine kinase inhibitors targeting ALK have significantly improved the prognosis of these patients. However, most patients experienced disease progression within a few years due to acquired resistance. Brigatinib is a second-generation ALK inhibitor effective in presence of several ALK mutations with demonstrated activity against central nervous system metastases. Currently, brigatinib is approved to treat ALK-positive metastatic NSCLC patients not previously treated with ALK inhibitors and patients who have progressed on or are intolerant to crizotinib. In this review, we provide a summary of results from clinical trials involving brigatinib, and we discuss its possible role in the management of ALK-positive NSCLC in the following years.


A deterministic compressive sensing model for bat biosonar.
The big brown bat (Eptesicus fuscus) uses frequency modulated (FM) echolocation calls to accurately estimate range and resolve closely spaced objects in clutter and noise. They resolve glints spaced down to 2 μs in time delay which surpasses what traditional signal processing techniques can achieve using the same echolocation call. The Matched Filter (MF) attains 10-12 μs resolution while the Inverse Filter (IF) achieves higher resolution at the cost of significantly degraded detection performance. Recent work by Fontaine and Peremans [J. Acoustic. Soc. Am. 125, 3052-3059 (2009)] demonstrated that a sparse representation of bat echolocation calls coupled with a decimating sensing method facilitates distinguishing closely spaced objects over realistic SNRs. Their work raises the intriguing question of whether sensing approaches structured more like a mammalian auditory system contains the necessary information for the hyper-resolution observed in behavioral tests. This research estimates sparse echo signatures using a gammatone filterbank decimation sensing method which loosely models the processing of the bat's auditory system. The decimated filterbank outputs are processed with [script-l](1) minimization. Simulations demonstrate that this model maintains higher resolution than the MF and significantly better detection performance than the IF for SNRs of 5-45 dB while undersampling the return signal by a factor of six.


Immune Response to Borrelia: Lessons from Lyme Disease Spirochetes.
The mammalian host responds to infection with Borrelia spirochetes through a highly orchestrated immune defense involving innate and adaptive effector functions aimed toward limiting pathogen burdens, minimizing tissue injury, and preventing subsequent reinfection. The evolutionary adaptation of Borrelia spirochetes to their reservoir mammalian hosts may allow for its persistence despite this immune defense. This review summarizes our current understanding of the host immune response to B. burgdorferi sensu lato, the most widely studied Borrelia spp. and etiologic agent of Lyme borreliosis. Pertinent literature will be reviewed with emphasis on in vitro, ex vivo and animal studies that influenced our understanding of both the earliest responses to B. burgdorferi as it enters the mammalian host and those that evolve as spirochetes disseminate and establish infection in multiple tissues. Our focus is on the immune response of inbred mice, the most commonly studied animal model of B. burgdorferi infection and surrogate for one of this pathogen's principle natural reservoir hosts, the white-footed deer mouse. Comparison will be made to the immune responses of humans with Lyme borreliosis. Our goal is to provide an understanding of the dynamics of the mammalian immune response during infection with B. burgdorferi and its relation to the outcomes in reservoir (mouse) and non-reservoir (human) hosts.


Convenient consumption: a critical qualitative inquiry into the gambling practices of younger women in Australia.
There are a range of stereotypes and assumptions associated with women's gambling behaviours. While researchers have demonstrated that the practices associated with women's gambling are changing and becoming increasingly normalized, there is a limited understanding of how younger women ascribe meanings to these practices. This study explored the gambling practices of younger women. Forty-one women (20-40 years) participated in qualitative telephone interviews. Participants were asked open-ended questions about personal engagement in gambling, including experiences of gambling, gambling engagement, and experiences with different gambling products and environments. Data interpretation was guided by reflexive thematic analysis. Three themes were constructed from the data: (i) gambling infrastructures, including both products and the embedding of gambling in community environments, contributed to the convenient and regular consumption of gambling, with gambling easy to access and engage with; (ii) social networks and intergenerational gambling practices impacted the perceived social value and competencies related to gambling; and (iii) technology facilitated new gambling practices, routinizing gambling behaviours through automation and building perceived competencies with a range of gambling products. Gambling regulation and public health responses to gambling often focus on either individual behaviours or product characteristics. This study suggests that this focus is too narrow and excludes important influences on younger women's gambling practices, which include the infrastructure that supports the provision and consumption of gambling products. Public health research, policy and practice must consider the full range of determinants that may contribute to the initiation and continuation of gambling in younger women.


Exonuclease III-assisted signal amplification strategy for sensitive fluorescence detection of polynucleotide kinase based on poly(thymine)-templated copper nanoparticles.
A sensitive and label-free fluorometric method has been developed for the determination of polynucleotide kinase (PNK) activity, by employing exonuclease III (Exo III)-assisted cyclic signal amplification and poly(thymine)-templated copper nanoparticles (polyT-CuNPs). In the presence of PNK, cDNA with 5'-hydroxyl termini was phosphorylated and then hybridized with tDNA to form the cDNA/tDNA duplex, which subsequently triggered the λ exonuclease cleavage reaction, eventually resulting in the release of tDNA. The released tDNA could unfold the hairpin structure of HP DNA to generate partially complementary duplex (tDNA/HP DNA), wherein the HP DNA possessed T-rich sequences (T30) and tDNA recognition sequence. With the help of Exo III digestion, the tDNA was able to initiate the cycle for the generation of T-rich sequences, the template for the formation of fluorescent CuNPs. Conversely, the cDNA could not be cleaved by λ exonuclease without PNK and individual HP DNA could not be hydrolyzed by Exo III. The T-rich sequence was caged in HP DNA, resulting in a weak fluorescence signal. Under optimized conditions, the fluorescence intensity was linearly correlated to a concentration range of 0.001 to 1 U mL-1 with a low detection limit of 2 × 10-4 U mL-1. Considering the intriguing analytical performance, this approach could be explored to screen T4 PNK inhibitors and hold promising applications in drug discovery and disease therapy.


Physical Inactivity Among Adults Aged 50 Years and Older - United States, 2014.
Physical activity can help delay, prevent, or manage many of the chronic diseases for which adults aged ≥50 years are at risk (1-3). These diseases can impact the length and quality of life, as well as the long-term ability to live independently.* All adults aged ≥50 years, with or without chronic disease, gain health benefits by avoiding inactivity (2,3). To examine the prevalence of inactivity by selected demographic characteristics and chronic disease status in mid-life and older adults, CDC analyzed data on adults aged ≥50 years from the 2014 Behavioral Risk Factor Surveillance System (BRFSS). Overall, 27.5% of adults aged ≥50 years reported no physical activity outside of work during the past month. Inactivity prevalence significantly increased with increasing age and was 25.4% among adults aged 50-64 years, 26.9% among those aged 65-74 years, and 35.3% among those aged ≥75 years. Inactivity prevalence was significantly higher among women than men, among Hispanics and non-Hispanic blacks than among non-Hispanic whites, and among adults who reported ever having one or more of seven selected chronic diseases than among those not reporting one. Inactivity prevalence significantly increased with decreasing levels of education and increasing body mass index. To help adults with and without chronic disease start or maintain an active lifestyle, communities can implement evidence-based strategies, such as creating or enhancing access to places for physical activity, designing communities and streets to encourage physical activity, and offering programs that address specific barriers to physical activity.


Genetic analysis of the antifungal activity of a soilborne Pseudomonas aureofaciens strain.
Pseudomonas aureofaciens Q2-87 produces the antibiotic 2,4-diacetophloroglucinol (Phl), which inhibits Gaeumannomyces graminis var. tritici and other fungi in vitro. Strain Q2-87 also provides biological control of take-all, a root disease of wheat caused by this fungus. To assess the role of Phl in the antifungal activity of strain Q2-87, a genetic analysis of antibiotic production was conducted. Two mutants of Q2-87 with altered antifungal activity were isolated by site-directed mutagenesis with Tn5. One mutant, Q2-87::Tn5-1, did not inhibit G. graminis var. tritici in vitro and did not produce Phl. Two cosmids were isolated from a genomic library of the wild-type strain by probing with the mutant genomic fragment. Antifungal activity and Phl production were coordinately restored in Q2-87::Tn5-1 by complementation with either cosmid. Mobilization of one of these cosmids into two heterologous Pseudomonas strains conferred the ability to synthesize Phl and increased their activity against G. graminis var. tritici, Pythium ultimum, and Rhizoctonia solani in vitro. Subcloning and deletion analysis of these cosmids identified a 4.8-kb region which was necessary for Phl synthesis and antifungal activity.


Breast cancer or metastasis? An unusual case of metastatic malignant pleural mesothelioma to the breast.
Metastases to the breast from extramammary malignancies are very rare, and ruling out the diagnosis of primary breast tumor is important in order to decide on clinical management and predict prognosis. Clinical examination revealed in a 49-year-old hairdresser a 3-cm hard lump adherent to the underlying layers in the right breast. Trucut biopsy was performed. Histology showed a solid proliferation of medium-sized neoplastic polygonal cells. Immunohistochemical analysis showed tumor cells diffusely positive for cytokeratin 8/18 and calretinin and focally positive for cytokeratin 5/6 and Wilms' tumor 1, e-cadherin, and human bone marrow endothelial-1. Estrogen receptors and progesterone receptors were negative. The final diagnosis was metastatic epithelioid malignant pleural mesothelioma. Immunohistochemistry is an important tool for a conclusive diagnosis of malignant pleural mesothelioma. Owing to the degree of histological and immunohistochemical overlap, a high level of clinical suspicion is essential in order to avoid unnecessary mutilating surgery.


Mammographic screening: case-control studies.
The case-control design can be used to evaluate the benefit of cancer screening programmes. This paper outlines the main methodological features of the case-control design in this context, and indicates some potential biases. It also reviews the existing case-control literature on mammographic screening. Case-control studies consistently indicate a reduction of approximately 50% in breast cancer mortality associated with mammography. This result indicates greater benefit than shown in randomised trials; however, one should recognise that trials indicate effectiveness whereas case-control studies indicate efficacy. The two types of evidence are broadly compatible when one allows for screening non-compliance and contamination in the randomised trials. The case-control evidence supports and is consistent with the findings of randomised trials of mammography. Effectiveness estimates from trials indicate the benefit of screening to the population as a whole, and are pertinent to the public policy debate as to the value of offering screening. In contrast, case-control studies indicate benefit to actual screening participants. As such, case-control estimates of efficacy are appropriate for individual decision-making by women about their use of mammography when it is potentially available to them.


ASSESSMENT OF THE NUTRITIONAL STATUS OF ADOLESCENTS IN A TERTIARY HEALTH FACILITY IN RIVERS STATE, NIGERIA.
Adolescence is a peculiar period of physical, sexual, psychosocial and cognitive development geared towards achieving their adult potentials. Adolescents are prone to malnutrition from unfavourable sociodemographic variables and sub-optimal nutrition. The study aimed to assess the nutritional status and the sociodemographic variables associated with malnutrition among adolescents in a tertiary health facility in Rivers State, Nigeria. A cross-sectional study was carried out among 10-17-year-old adolescents in the Paediatric Out-patient Clinic, River State University Teaching Hospital. A proforma was used to collect sociodemographic data from them before taking their anthropometric measurements. BMI- for-age Z scores were calculated using the WHO anthropometric calculator and nutritional status determined using the WHO classification. Univariate and multivariate data analysis was done using SPPS version 23. Statistical significance was set at P value < 0.05. A total of 131 adolescents aged 10 to 17 years, with 72(55%) females participated in the study. Majority were urban dwellers, 61.1% attended private educational institutions, 46% were from middle socioeconomic class and 40(30.5%) from families with >4 children. The median weight and height of the study population were 47kg and 157cm respectively. The mean BMI-for-age Z score was -0.126±0.318. Twenty-six (19.9%) adolescents were overweight/obese while 9(6.9%) were too thin (wasting). Adolescents aged 10-13 years were significantly more overweight/obese than others while wasting was more prevalent among adolescents dwelling in rural communities, in primary schools and living in households with >4 children (p < 0.05). There was a 29-fold increased risk of wasting among children living in households with >4 children (P=0.002). The study highlights two extremes of malnutrition among adolescents with overnutrition more than two times that of undernutrition. Adolescents living in large households are at risk of undernutrition.


Relation of Body Mass Index to Development of Atrial Fibrillation in Hispanics, Blacks, and Non-Hispanic Whites.
No previous studies have examined the interaction between body mass index (BMI) and race/ethnicity with the risk of atrial fibrillation (AF). We retrospectively followed 48,323 persons free of AF (43% Hispanic, 37% black, and 20% white; median age 60 years) for subsequent incident AF (ascertained from electrocardiograms). BMI categories included very severely underweight (BMI <15 kg/m2), severely underweight (BMI 15.1 to 15.9 kg/m2), underweight (BMI 16 to 18.4 kg/m2), normal (BMI 18.5 to 24.9 kg/m2), overweight (BMI 25.0 to 29.9 kg/m2), moderately obese (BMI 30 to 34.9 kg/m2), severely obese (BMI 35 to 39.9 kg/m2), and very severely obese (BMI >40 kg/m2). Cox regression analysis controlled for baseline covariates: heart failure, gender, age, treatment for hypertension, diabetes, PR length, systolic blood pressure, left ventricular hypertrophy, socioeconomic status, use of β blockers, calcium channel blockers, and digoxin. Over a follow-up of 13 years, 4,744 AF cases occurred. BMI in units of 10 was associated with the development of AF (adjusted hazard ratio 1.088, 95% confidence interval 1.048 to 1.130, p <0.01). When stratified by race/ethnicity, non-Hispanic whites compared with blacks and Hispanics had a higher risk of developing AF, noted in those whom BMI classes were overweight to severely obese. In conclusion, our study demonstrates that there exists a relation between obesity and race/ethnicity for the development of AF. Non-Hispanic whites had a higher risk of developing AF compared with blacks and Hispanics.


CRKL amplification is rare as a mechanism for acquired resistance to kinase inhibitors in lung cancers with epidermal growth factor receptor mutation.
Epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) often provide dramatic responses in lung cancer patients with somatic EGFR mutation. However, acquired resistance to the drugs usually emerges within a few years. EGFR T790M secondary mutation, MET gene amplification, and transformation to small cell lung cancer are well-validated mechanisms that underlie acquisition of resistance to EGFR-TKIs. In addition, many molecular aberrations have been reported as candidates for mechanisms of acquired resistance to EGFR-TKIs. Amplification of the CRKL gene was reportedly observed in 1 of 11 lung cancer patients with EGFR mutations who acquired resistance to EGFR-TKI. This study is the first report, to our knowledge, that validated the role of CRKL gene amplification as a mechanism for acquisition of resistance to EGFR-TKIs. We analyzed CRKL gene copy numbers, using a quantitative real-time PCR method, in 2 in vitro acquired-resistance cell-line models: 11 clinical samples from patients who developed acquired resistance to EGFR-TKIs, and 39 tumor specimens obtained from 7 autopsy patients whose cancers acquired resistance to EGFR-TKIs. Mutational status of EGFR codon 790 and copy numbers for the MET gene were also determined. In analysis for in vitro models, CRKL gene copy numbers were identical between EGFR-TKI-sensitive parental cells and their acquired resistant descendant cells. In addition, we found no clinical tumor specimens with acquired EGFR-TKI resistance to harbor amplified CRKL genes. These results indicate that CRKL gene amplification is rare in acquisition of resistance to EGFR-TKIs in lung cancer patients with EGFR mutations.


Does the Presence of Blood in the Catheter or the Degree of Difficulty of Embryo Transfer Affect Live Birth?
The technique used for embryo transfer (ET) can affect implantation. Prior research that evaluated the effect of postprocedural blood of the transfer catheter tip have yielded mixed results, and it is unclear whether this is actually a marker of difficulty of the transfer. Our objective was to estimate the effect of blood at the time of ET and the difficulty of ET on live birth rates (LBR). This retrospective cohort study utilized generalized estimating equations (GEEs) with nesting for repeated cycles for all analyses. Univariate modeling was performed and a final multivariate (adjusted) GEE model accounted for all significant confounders. Embryo transfers were subjectively graded (easy, medium, or hard) by a physician at the time of transfer. The presence of blood at ET was associated with more difficult ETs, retained embryos, and presence of mucous in the catheter. In the univariate analysis, ET with blood was not associated with live birth, while the degree of difficulty for ET had a negative impact on LBR. In the final multivariate GEE model, which accounts for repeated cycles from a patient, the only factors associated with an increased LBR were the degree of difficulty of the ET, female age, and blastocyst transfer. After controlling for confounding variables, the presence of blood in the transfer catheter was not associated with the likelihood of pregnancy and thus was not an independent predictor of cycle outcome. This indicates that the difficulty of the transfer itself was a strong negative predictor of pregnancy.


HIV-1 TAT inhibits microglial phagocytosis of Abeta peptide.
Human immunodeficiency virus (HIV)-associated dementia (HAD) is a subcortical neuropsychiatric syndrome that has increased in prevalence in the era of highly active antiretroviral therapy (HAART). Several studies demonstrated increased amyloidosis in brains of HIV patients and suggested that there may be a significant number of long-term HIV survivors with co-morbid Alzheimer's disease (AD) in the future. We show HIV-1 Tat protein inhibits microglial uptake of Abeta1-42 peptide, a process that is enhanced by interferon-gamma (IFN-gamma) and rescued by the STAT1 inhibitor (-)-epigallocatechin-3-gallate (EGCG). It is hypothesized that reduced Abeta uptake occurs through IFN-gamma mediated STAT1 activation. This process promotes a switch from a phagocytic to an antigen presenting phenotype in microglia through activation of class II transactivator (CIITA). Additionally, we show that HIV-1 Tat significantly disrupts apolipoprotein-3 (Apo-E3) promoted microglial Abeta uptake. As Tat has been shown to directly interact with the low density lipoprotein (LRP) receptor and thus inhibit the uptake of its ligands including apolipoprotein E4 (Apo-E4) and Abeta peptide in neurons, we further hypothesize that a similar inhibition of LRP may occur in microglia. Future studies will be required to fully characterize the mechanisms underlying IFN-gamma enhancement of HIV-1 Tats disruption of microglial phagocytosis of Abeta and Apo-E3.


Case Study: Muscle Atrophy, Hypertrophy, and Energy Expenditure of a Premier League Soccer Player During Rehabilitation From Anterior Cruciate Ligament Injury.
Maintaining muscle mass and function during rehabilitation from anterior cruciate ligament injury is complicated by the challenge of accurately prescribing daily energy intakes aligned to energy expenditure. Accordingly, we present a 38-week case study characterizing whole body and regional rates of muscle atrophy and hypertrophy (as inferred by assessments of fat-free mass from dual-energy X-ray absorptiometry) in a professional male soccer player from the English Premier League. In addition, in Week 6, we also quantified energy intake (via the remote food photographic method) and energy expenditure using the doubly labeled water method. Mean daily energy intake (CHO: 1.9-3.2, protein: 1.7-3.3, and fat: 1.4-2.7 g/kg) and energy expenditure were 2,765 ± 474 and 3,178 kcal/day, respectively. In accordance with an apparent energy deficit, total body mass decreased by 1.9 kg during Weeks 1-6 where fat-free mass loss in the injured and noninjured limb was 0.9 and 0.6 kg, respectively, yet, trunk fat-free mass increased by 0.7 kg. In Weeks 7-28, the athlete was advised to increase daily CHO intake (4-6 g/kg) to facilitate an increased daily energy intake. Throughout this period, total body mass increased by 3.6 kg (attributable to a 2.9 and 0.7 kg increase in fat free and fat mass, respectively). Our data suggest it may be advantageous to avoid excessive reductions in energy intake during the initial 6-8 weeks post anterior cruciate ligament surgery so as to limit muscle atrophy.


Caregiver-identified needs and barriers to care in Parkinson's disease.
Perceptions of service needs and barriers to care among caregivers of individuals with Parkinson's disease have not been well explored. The purpose of this study was to assess caregiver perceptions of their own and patients' medical and supportive service needs. An online and paper survey was disseminated to a sample of caregivers (n = 66) of individuals with Parkinson's disease. Although caregivers reported positive quality of life and adjustment to caregiving, nearly half of the sample endorsed feeling stressed about caregiving. Caregivers reported that services for symptom management, coping with changes in lifestyle, future planning, relationships, and cognition, and wellness strategies were most needed. Reported barriers to patients accessing care included limited service availability and a lack of insurance coverage for services. These findings suggest a need to improve access to services for patients and increased efforts to promote caregiver wellness at movement disorder specialty clinics.


Dynamic versus static analyses of lifting a box from the floor.
Lifting objects from below knee height has been implicated as a source of low back pain and injury. Static models have often been used to assess forces produced in the lumbar region by lifting; however, inertial forces generated by acceleration may be significant. Therefore, the goal of this investigation was to assess differences between static and dynamic analysis methods. Sagittal plane kinematics were collected on 21 men and 22 women of college age while lifting a milk crate (men = 25 kg, women = 15 kg) from the floor to standing knuckle height on level as well as sloped (facing uphill and downhill at 10 degrees and 20 degrees) ground conditions. Both static and dynamic top-down inverse models were utilized to assess net muscular moments at L5/S1 as well as the posture of the person at the time of static max (TSM) and dynamic max (TDM) moments. The TDM moment was significantly later than the TSM in the level through uphill conditions (p < 0.001). The dynamic max moment was significantly greater than the static max moment in all conditions (p < 0.001). Torso angles at TSM exhibited a significantly greater forward lean (by < 2 degrees) in the level through uphill conditions (p < 0.001). Overall low-back curvature, hip angles, knee angles, and ankle angles were not affected by the type of model (p > 0.05), though several minor differences occurred at conditions other than the level (most dramatic in the downhill 20 degrees condition). Therefore, if moments are of interest, a dynamic model should be utilized. However, body position is very similar at TSM and TDM.


Effects of a fully magnetically levitated centrifugal-flow or axial-flow left ventricular assist device on von Willebrand factor: A prospective multicenter clinical trial.
Increased shear stress conferred upon the circulation by continuous-flow pumps is associated with hemocompatibility-related adverse events, principally bleeding within the gastrointestinal system, and linked to the degradation of high-molecular-weight multimers (HMWMs) of von Willebrand factor (vWF). We evaluated the structure and functional characteristics of vWF HMWMs in patients with the fully magnetically levitated centrifugal-flow HeartMate 3 (HM3) and the continuous axial-flow HeartMate II (HMII) pump. Findings were correlated with bleeding events. In a prospective, multicenter, comparative cohort study, 60 patients from the Multicenter Study of MagLev Technology in Patients Undergoing Mechanical Circulatory Support Therapy With HeartMate 3 Continued Access Protocol (NCT02892955) with an HM3 pump were compared with 30 randomly selected HMII patients from the PREVENtion of HeartMate II Pump Thrombosis study (NCT02158403) biobank. The primary end point was the difference in the normalized vWF HMWM ratio (ratio of the HMWMs to the intermediate- and low-molecular-weight multimers, normalized to pooled plasma from healthy volunteers) between the HM3 and the HMII pump at 90 days after implantation. Assay tests for vWF activity, vWF antigen, vWF activity to antigen ratio, coagulation factor VIII activity, and ADAMTS13 activity were measured by using standard protocols. Differences in these markers were compared in the context of clinical characteristics and correlated with adjudicated bleeding events within the HM3 group. Of 51 and 29 evaluable patients in the HM3 and HMII arms, respectively, those implanted with the HM3 pump exhibited greater preservation of the vWF HMWM ratio than those with the HMII pump at 90 days after implantation (54.1% vs 42.4%, p < 0.0001). Laboratory values for all vWF assays (antigen, activity, and coagulation factor VIII activity) remained within the normal functional range with no significant differences observed between the pumps at 90 days after implantation. At baseline, there was a decrease in the structural integrity of vWF HMWMs that correlated with increasing heart failure severity as measured by the Interagency Registry for Mechanically Assisted Circulatory Support profile. Multivariable modeling identified the HM3 pump as the only independent variable that determined post-implantation preservation of the structural integrity of vWF HMWMs. This prospective, multicenter comparative analysis study demonstrates that the fully magnetically levitated centrifugal-flow HM3 left ventricular assist device is associated with greater preservation of the structure of vWF HMWMs than the HMII mechanical bearing axial-flow pump.


Synergistic effects of ion transporter and MAP kinase pathway inhibitors in melanoma.
New therapies are required for melanoma. Here, we report that multiple cardiac glycosides, including digitoxin and digoxin, are significantly more toxic to human melanoma cells than normal human cells. This reflects on-target inhibition of the ATP1A1 Na(+)/K(+) pump, which is highly expressed by melanoma. MEK inhibitor and/or BRAF inhibitor additively or synergistically combined with digitoxin to induce cell death, inhibiting growth of patient-derived melanomas in NSG mice and synergistically extending survival. MEK inhibitor and digitoxin do not induce cell death in human melanocytes or haematopoietic cells in NSG mice. In melanoma, MEK inhibitor reduces ERK phosphorylation, while digitoxin disrupts ion gradients, altering plasma membrane and mitochondrial membrane potentials. MEK inhibitor and digitoxin together cause intracellular acidification, mitochondrial calcium dysregulation and ATP depletion in melanoma cells but not in normal cells. The disruption of ion homoeostasis in cancer cells can thus synergize with targeted agents to promote tumour regression in vivo.


The duplicated Saccharomyces cerevisiae gene SSM1 encodes a eucaryotic homolog of the eubacterial and archaebacterial L1 ribosomal proteins.
A previously unknown Saccharomyces cerevisiae gene, SSM1a, was isolated by screening for high-copy-number suppressors of thermosensitive mutations in the RNA14 gene, which encodes a component from the polyadenylation complex. The SSM1 a gene codes for a 217-amino-acid protein, Ssm1p, which is significantly homologous to eubacterial and archaebacterial ribosomal proteins of the L1 family. Comparison of the Ssm1p amino acid sequence with that of eucaryotic polypeptides with unknown functions reveals that Ssm1p is the prototype of a new eucaryotic protein family. Biochemical analysis shows that Ssm1p is a structural protein that forms part of the largest 60S ribosomal subunit, which does not exist in a pool of free proteins. SSM1 a is duplicated. The second gene copy, SSM1b, is functional and codes for an identical and functionally interchangeable Ssm1p protein. In wild-type cells, SSM1b transcripts accumulate to twice the level of SSM1a transcripts, suggesting that SSM1b is responsible for the majority of the Ssm1p pool. Haploid cells lacking both SSM1 genes are inviable, demonstrating that, in contrast with its Escherichia coli homolog, Ssm1p is an essential ribosomal protein. Deletion of the most expressed SSM1b gene leads to a severe decrease in the level of SSM1 transcript, associated with a reduced growth rate. Polysome profile analysis suggests that the primary defect caused by the depletion in Ssm1p is at the level of translation initiation.


Rostral anterior cingulate cortex activity and early symptom improvement during treatment for major depressive disorder.
In treatment trials for major depressive disorder (MDD), early symptom improvement is predictive of eventual clinical response. Clinical response may also be predicted by elevated pretreatment theta (4-7Hz) current density in the rostral anterior cingulate (rACC) and medial orbitofrontal cortex (mOFC). We investigated the relationship between pretreatment EEG and early improvement in predicting clinical outcome in 72 MDD subjects across three placebo-controlled treatment trials. Subjects were randomized to receive fluoxetine, venlafaxine, or placebo. Theta current density in the rACC and mOFC was computed with Low-Resolution Brain Electromagnetic Tomography (LORETA). An analysis of covariance examining week-8 Hamilton Depression Rating Scale (HamD) percent change, showed a significant effect of week-2HamD percent change, and a significant three-way interaction of week-2HamD percent change×treatment×rACC. Medication subjects with robust early improvement showed almost no relationship between rACC theta current density and final clinical outcome. However, in subjects with little early improvement, rACC activity showed a strong relationship with clinical outcome. The model examining the mOFC showed a trend in the three-way interaction. A combination of pretreatment rACC activity and early symptom improvement may be useful for predicting treatment response.


Giant Metastatic Breast Phyllodes Tumour with an Elusive Diagnosis: A Case Report and Literature Review.
The term phyllodes tumours, which account for less than 1% of breast neoplasms, describes a spectrum of heterogenous tumours with different clinical behaviours. Less than 30% present as metastatic disease. Complete surgical resection is the standard of care so that recurrence rates are reduced. The role of adjuvant chemotherapy or radiation therapy is controversial. Patients with metastatic disease have a median overall survival of around 30 months. The authors present the case of a 57-year-old woman with an exuberant left malignant phyllodes tumour with bilateral involvement, as well as lung and axillar metastasis. The patient underwent haemostatic radiation therapy and started palliative chemotherapy with doxorubicin, achieving partial response with significant improvement in quality of life. A posterior simple mastectomy revealed a small residual tumour. Metastatic malignant phyllodes tumours are rare, so therapeutic strategies rely on small retrospective studies and guidelines for soft tissue sarcoma. Palliative chemotherapy protocols include anthracycline-based regimens, either as monotherapy with doxorubicin or doxorubicin together with ifosfamide. With few treatment options, management of these patients must rely on a continuum of care. Phyllodes tumours are a rare type of breast neoplasm.The differential diagnosis of breast cancer should include phyllodes tumours.Accurate and rapid diagnosis is required.


Fatal gastrointestinal hemorrhage after a single dose of dabigatran.
Dabigatran (Pradaxa) is a new oral anticoagulant approved by the Food and Drug Administration (FDA), available internationally and indicated as an alternative to warfarin for the prevention of stroke and systemic embolism in patients with nonvalvular atrial fibrillation. Dabigatran does not require laboratory monitoring and its kinetics allow for a more rapid onset of action with a time to peak concentration of 1.25-1.5 h. We are reporting a fatality resulting from gastrointestinal bleeding after the ingestion of a single dose of dabigatran 150 mg. A 92-year-old man with a medical history of chronic obstructive pulmonary disease, hypothyroidism, and atrial flutter presented to the emergency department with complaints of weakness and rectal bleeding. He was seen by his Cardiologist the day before and was found to be in new atrial fibrillation. He was prescribed dabigatran 150 mg twice daily for anticoagulation therapy. He took one dose of dabigatran 150 mg at 2200 and woke up the following morning before 0900 with profuse rectal bleeding. The initial vital signs in the emergency department, approximately 11 h after ingestion, were heart rate 72 beats/min, blood pressure 62/30 mmHg, and lab work showed hemoglobin 9.9 g/dL, international normalization ratio (INR) 1.99, blood urea nitrogen (BUN) 66 mg/dL, and creatinine (SCr) 1.4 mg/dL (creatinine clearance (CrCl) 24.2 mL/min). He was resuscitated with intravenous fluids, two units of packed red blood cells, two units of fresh frozen plasma, platelets, and vitamin K 10 mg intravenously. He was also given an unknown dose of erythromycin early in his hospital stay. An actively bleeding gastric ulcer was discovered and treated with local epinephrine injections. Approximately 48 h after his exposure, he received an additional two units of blood to treat his decreasing blood pressure (98/41 mmHg). On day three, his hemoglobin and hematocrit were stable at 10 g/dL and 30%, INR 1.6, he was extubated and off vasoactive medications. Day six of hospitalization, he began having maroon stools, his hemoglobin decreased to 8.1 g/dL and his platelets to 81 × 1000/mcL. On day seven, the hemoglobin decreased to 6.4 mg/dL. Despite aggressive resuscitative efforts and supportive care, he died. This case demonstrates the potential of a single dose of dabigatran 150 mg to result in a fatal gastrointestinal hemorrhage. This patient was started on the maximum dose with a CrCl 33.9 mL/min and on admission CrCl 24.2 mL/min, suggesting underlying renal insufficiency.


Association of Mitochondrial DNA Copy Number With Cardiovascular Disease.
Mitochondrial dysfunction is a core component of the aging process and may play a key role in atherosclerotic cardiovascular disease. Mitochondrial DNA copy number (mtDNA-CN), which represents the number of mitochondria per cell and number of mitochondrial genomes per mitochondrion, is an indirect biomarker of mitochondrial function. To determine whether mtDNA-CN, measured in an easily accessible tissue (buffy coat/circulating leukocytes), can improve risk classification for cardiovascular disease (CVD) and help guide initiation of statin therapy for primary prevention of CVD. Prospective, population-based cohort analysis including 21 870 participants (20 163 free from CVD at baseline) from 3 studies: Cardiovascular Health Study (CHS), Atherosclerosis Risk in Communities Study (ARIC), and Multiethnic Study of Atherosclerosis (MESA). The mean follow-up was 13.5 years. The study included 11 153 participants from ARIC, 4830 from CHS, and 5887 from MESA. Analysis of the data was conducted from March 10, 2014, to January 29, 2017. Mitochondrial DNA-CN measured from buffy coat/circulating leukocytes. Incident CVD, which combines coronary heart disease, defined as the first incident myocardial infarction or death owing to coronary heart disease, and stroke, defined as the first nonfatal stroke or death owing to stroke. Of the 21 870 participants, the mean age was 62.4 years (ARIC, 57.9 years; MESA, 62.4 years; and CHS, 72.5 years), and 54.7% of participants were women. The hazard ratios for incident coronary heart disease, stroke, and CVD associated with a 1-SD decrease in mtDNA-CN were 1.29 (95% CI, 1.24-1.33), 1.11 (95% CI, 1.06-1.16), and 1.23 (95% CI, 1.19-1.26). The associations persisted after adjustment for traditional CVD risk factors. Addition of mtDNA-CN to the 2013 American College of Cardiology/American Heart Association Pooled Cohorts Equations for estimating 10-year hard atherosclerosis CVD risk was associated with improved risk classification (continuous net reclassification index, 0.194; 95% CI, 0.130-0.258; P < .001). Mitochondrial DNA-CN further improved sensitivity and specificity for the 2013 American College of Cardiology/American Heart Association recommendations on initiating statin therapy for primary prevention of ASCVD (net 221 individuals appropriately downclassified and net 15 individuals appropriately upclassified). Mitochondrial DNA-CN was independently associated with incident CVD in 3 large prospective studies and may have potential clinical utility in improving CVD risk classification.


Rif1 regulates initiation timing of late replication origins throughout the S. cerevisiae genome.
Chromosomal DNA replication involves the coordinated activity of hundreds to thousands of replication origins. Individual replication origins are subject to epigenetic regulation of their activity during S-phase, resulting in differential efficiencies and timings of replication initiation during S-phase. This regulation is thought to involve chromatin structure and organization into timing domains with differential ability to recruit limiting replication factors. Rif1 has recently been identified as a genome-wide regulator of replication timing in fission yeast and in mammalian cells. However, previous studies in budding yeast have suggested that Rif1's role in controlling replication timing may be limited to subtelomeric domains and derives from its established role in telomere length regulation. We have analyzed replication timing by analyzing BrdU incorporation genome-wide, and report that Rif1 regulates the timing of late/dormant replication origins throughout the S. cerevisiae genome. Analysis of pfa4Δ cells, which are defective in palmitoylation and membrane association of Rif1, suggests that replication timing regulation by Rif1 is independent of its role in localizing telomeres to the nuclear periphery. Intra-S checkpoint signaling is intact in rif1Δ cells, and checkpoint-defective mec1Δ cells do not comparably deregulate replication timing, together indicating that Rif1 regulates replication timing through a mechanism independent of this checkpoint. Our results indicate that the Rif1 mechanism regulates origin timing irrespective of proximity to a chromosome end, and suggest instead that telomere sequences merely provide abundant binding sites for proteins that recruit Rif1. Still, the abundance of Rif1 binding in telomeric domains may facilitate Rif1-mediated repression of non-telomeric origins that are more distal from centromeres.


MicroRNA-21 promotes Th17 differentiation and mediates experimental autoimmune encephalomyelitis.
Accumulation of IL-17-producing Th17 cells is associated with the development of multiple autoimmune diseases; however, the contribution of microRNA (miRNA) pathways to the intrinsic control of Th17 development remains unclear. Here, we demonstrated that miR-21 expression is elevated in Th17 cells and that mice lacking miR-21 have a defect in Th17 differentiation and are resistant to experimental autoimmune encephalomyelitis (EAE). Furthermore, we determined that miR-21 promotes Th17 differentiation by targeting and depleting SMAD-7, a negative regulator of TGF-β signaling. Moreover, the decreases in Th17 differentiation in miR-21-deficient T cells were associated with defects in SMAD-2/3 activation and IL-2 suppression. Finally, we found that treatment of WT mice with an anti-miR-21 oligonucleotide reduced the clinical severity of EAE, which was associated with a decrease in Th17 cells. Thus, we have characterized a T cell-intrinsic miRNA pathway that enhances TGF-β signaling, limits the autocrine inhibitory effects of IL-2, and thereby promotes Th17 differentiation and autoimmunity.


Risk of breast cancer and total malignancies in rheumatoid arthritis patients undergoing TNF-α antagonist therapy: a meta-analysis of randomized control trials.
Interest exits in whether TNF-alpha antagonists increase the risk of breast cancer and total malignancies in patients with rheumatoid arthritis (RA). To analyze the risk of malignancies, especially breast cancer, in patients with RA enrolled in randomized control trials (RCTs). A systematic literature search for RCTs from 1 January 1998 to 1 July 2013 from online databases, such as PubMed, WILEY, EMBASE, ISI web of knowledge and Cochrane Library was conducted. Studies included RCTs that compared the safety of at least one dose of the five TNF-α antagonists with placebo or methotrexate (MTX) (or TNF-α antagonists plus MTX vs placebo plus MTX) in RA patients for more than 24 weeks and imported all the references into document management software EndNotex6. Two independent reviewers selected studies and extracted the data about study design, patients' characteristics and the type, number of all malignancies. 28 RCTs from 34 records with 11,741 patients were analyzed. Of the total, 97 developed at least one malignancy during the double-blind trials, and breast cancer was observed in 17 patients (17.5% of total malignancies). However, there was no statistically significant increased risk observed in either the per protocol (PP) model (OR 0.65, 95%CI [0.22, 1.93]) or the modified intention to treat (mITT) model (OR 0.75, 95%CI [0.25, 2.21]). There were also no significant trend for increased risk of total malignancies on anti-TNF-α therapy administered at approved doses in either model (OR, 1.06, 95%CI [0.64, 1.75], and OR, 1.30, 95%CI [0.80, 2.14], respectively). As to the two models, modified intention to treat model analysis led to higher estimation than per protocol model analysis. This study did not find a significantly increased risk of breast cancer and total malignancies in adults RA patients treated with TNF-α antagonists at approved doses. However, it cannot be ignored that more patients developed malignancies with TNF-α antagonists therapy compared with patients with placebo or MTX, in spite of the lack of statistical significance, so that more strict clinical trials and long-term follow-up are needed, and both mITT and PP analyses should be used in such safety analyses.


A lyoluminescent tissue-equivalent dosemeter for pion therapy beams.
The light emitted when pion irradiated glucose is dissolved in luminol solution has been found to be proportional to the pion beam depth dose distribution in water as determined by a TE ionisation chamber. The lyoluminescence of glucose overlaps very closely with the response profile of the ionisation chamber to the 170 MeV/c pi--mesons giving a Bragg peak to plateau ratio of 3:1. In comparison, the thermoluminescence response of LiF (TLD-700) to pions has been found to deviate significantly from this ratio. The close tissue equivalence of glucose, non-toxicity and its excellent lyoluminescent retention properties are important advantages over currently used dosemeters in clinical pion therapy, especially when direct in vivo measurements are considered. Pion doses ranging between 0.5 Gy (50 rad) and 30 Gy (3000 rad) were measured with an accuracy +/- 5% and reproducibility 3-5%.


Acute Coronary Syndromes in the Elderly.
The clinical evidence for treatment of acute coronary syndrome (ACS) in the elderly is less robust than in patients younger than 75 years. The elderly have the highest incidence of cardiovascular disease and frequently present with ACS. This number can be expected to increase over time because society is aging. Older adults often sustain unfavorable outcomes from ACS because of atypical presentation and delay in recognition. In addition, elderly patients commonly do not receive optimal guideline-directed ACS treatment. Owing to their high baseline risk of ischemic complications, the elderly also fare worse even with optimal ACS treatment as they frequently have more complex coronary disease, more comorbidities, less cardiovascular reserve, and a higher risk of treatment complications. They are also subjected to a broader range of pharmacologic treatment. Treatment complications can be mitigated to some extent by meticulous dose adjustment of antithrombotic and adjunctive therapies. While careful transitions of care and appropriate utilization of post-discharge secondary preventive measures are important in ACS patients of all ages, the elderly are more vulnerable to system errors and thus deserve special attention from the clinician.


Human papillomavirus and the landscape of secondary genetic alterations in oral cancers.
Human papillomavirus (HPV) is a necessary but insufficient cause of a subset of oral squamous cell carcinomas (OSCCs) that is increasing markedly in frequency. To identify contributory, secondary genetic alterations in these cancers, we used comprehensive genomics methods to compare 149 HPV-positive and 335 HPV-negative OSCC tumor/normal pairs. Different behavioral risk factors underlying the two OSCC types were reflected in distinctive genomic mutational signatures. In HPV-positive OSCCs, the signatures of APOBEC cytosine deaminase editing, associated with anti-viral immunity, were strongly linked to overall mutational burden. In contrast, in HPV-negative OSCCs, T>C substitutions in the sequence context 5'-ATN-3' correlated with tobacco exposure. Universal expression of HPV E6*1 and E7 oncogenes was a sine qua non of HPV-positive OSCCs. Significant enrichment of somatic mutations was confirmed or newly identified in PIK3CA, KMT2D, FGFR3, FBXW7, DDX3X, PTEN, TRAF3, RB1, CYLD, RIPK4, ZNF750, EP300, CASZ1, TAF5, RBL1, IFNGR1, and NFKBIA Of these, many affect host pathways already targeted by HPV oncoproteins, including the p53 and pRB pathways, or disrupt host defenses against viral infections, including interferon (IFN) and nuclear factor kappa B signaling. Frequent copy number changes were associated with concordant changes in gene expression. Chr 11q (including CCND1) and 14q (including DICER1 and AKT1) were recurrently lost in HPV-positive OSCCs, in contrast to their gains in HPV-negative OSCCs. High-ranking variant allele fractions implicated ZNF750, PIK3CA, and EP300 mutations as candidate driver events in HPV-positive cancers. We conclude that virus-host interactions cooperatively shape the unique genetic features of these cancers, distinguishing them from their HPV-negative counterparts.


Bivariate linkage between acylation-stimulating protein and BMI and high-density lipoproteins.
Given the importance of visceral adiposity in the metabolic syndrome, whether levels of adipokines have shared genetic effects (pleiotropy) with aspects of the metabolic syndrome should be addressed. Acylation-stimulating protein (ASP), an adipose-derived protein, influences lipid metabolism, obesity, and glucose use. Therefore, our objective was to examine the genetic regulation of ASP and associated pleiotropic effects. We assayed serum ASP levels in 435 Mexican Americans participating in the San Antonio Family Heart Study and performed univariate and bivariate variance components analysis. Additive genetic heritability of ASP was 26% (p = 0.0004). Bivariate genetic analysis detected significant genetic correlations between ASP and several lipid measures but not between ASP and adiposity or diabetes measures. We detected two potential quantitative trait loci influencing ASP levels. The strongest signal was on chromosome 17 near marker D17S1303 [log of the odds ratio (LOD) = 2.7]. The signal on chromosome 15 reached its peak near marker D15S641 (LOD = 2.1). Both signals localize in regions reported to harbor quantitative trait loci influencing obesity and lipid phenotypes in this population. Bivariate linkage analysis yielded LODs of 4.7 for ASP and BMI on chromosome 17 and 3.2 for ASP and high-density lipoprotein2a on chromosome 15. Given these findings, there seems to be a significant genetic contribution to variation in circulating levels of ASP and an interesting pattern of genetic correlation (i.e., pleiotropy) with other risk factors associated with the metabolic syndrome.


Effect of ketoconazole 1% and 2% shampoos on severe dandruff and seborrhoeic dermatitis: clinical, squamometric and mycological assessments.
Ketoconazole (KET) is active to control dandruff and seborrhoeic dermatitis. Objective assessments comparing the 1% and 2% shampoo formulations are scant. This open, randomized parallel-group trial was carried out to differentiate the effectiveness of KET 1% and 2% in severe dandruff and seborrhoeic dermatitis. A total of 66 patients with severe dandruff or seborrhoeic dermatitis were randomized to each of the two groups. A 2-week run-in phase was followed by a 4-week treatment phase, in turn followed by a 4-week follow-up. The efficacy of treatments was evaluated by combining squamometry X, Malassezia spp. counts and clinical assessments. After 2 and 4 weeks of treatment, KET 2% was significantly superior over KET 1% (p < 0.001) for decreasing both in flakiness and Malassezia density from baseline. The same trend was observed in the mean change from baseline in the overall dandruff severity score. Only 6 mild adverse events were reported. During follow-up KET 2% showed a trend to fewer relapses than KET 1%. KET 2% had superior efficacy compared to KET 1% in the treatment of severe dandruff and scalp seborrhoeic dermatitis. Biometrological evaluations were correlated with the clinical improvements and therefore useful to incorporate in future dandruff studies.


Adenosine 5'-O-(3-thiotriphosphate) can support the formation of an initiation complex between the DNA polymerase III holoenzyme and primed DNA.
Adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S) will substitute for ATP in the formation of an initiation complex between the DNA polymerase III holoenzyme of Escherichia coli and primed DNA. The initiation complex formed in the presence of ATP gamma S between the DNA polymerase III holoenzyme and single-stranded DNA-binding protein-encoated primed M13 Gori DNA is stabile and isolable by gel filtration at room temperature. Upon addition of the four required deoxynucleoside triphosphates, this complex is rapidly converted to the duplex replicative form without dissociation of the polymerase. Initiation complexes formed in the presence of either ATP gamma S or ATP are indistinguishable by their resistance to antibody directed against the beta subunit of the holoenzyme and by their ability to elongate without further activation. A 2-fold difference was observed, however, in both the extent of initiation complex formation and in the dissociation of initiation complexes once formed. This difference is discussed in the light of previous proposals regarding a dimeric polymerase capable of replicating both strands at a replication fork concurrently.


Castor bean toxicity re-examined: a new perspective.
Commonly used references present a very gloomy prognosis for castor seed ingestion. This appears to be based chiefly on extrapolations from laboratory animal studies of toxicity of the component, ricin. This is not in concordance with actual experience when actual case report experience is sought out and examined in detail. The literature has been confused by recursive references among secondary sources and mortality statistics have been distorted by reliance on data generated nearly a century ago which do not accurately reflect the capabilities of modern medical management. This is elucidated by actually tracking down and studying difficult-to-find original case reports from the early decades of the 20th century. It is time to reappraise our estimates of the hazard of ingesting castor seeds in the light of present day medical capabilities.


The number of sex-factors per chromosome in Escherichia coli.
A substituted sex-factor of Escherichia coli, F'8 gal, was transferred to Proteus mirabilis by conjugation. The DNA of the episome was partially purified from Proteus DNA by preparative equilibrium centrifugation in caesium chloride, and by a bulk method using hydroxyapatite. The buoyant density of the episomal DNA is 1.707, corresponding to a (G+C) content of 47%. By optical renaturation the genetic complexity of the episomal DNA was found to be 76x10(6) daltons. RNA was synthesized in vitro by using the episomal DNA as template. By hybridizing this RNA with DNA extracted from E. coli carrying F'8 gal, it is shown that the number of copies of the episome per replicating chromosome is close to two during exponential growth. The episome makes up about 4.4% of the total DNA of the growing cells. The activities of galactokinase and galactose 1-phosphate uridylyl-transferase in cells with and without episomal and chromosomal gal genes were found to be proportional to the number of gal genes present, when the cells were induced with d-fucose, but not when they were induced with d-galactose.


Photopolymerization of Macroscale Black 3D Objects Using Near-Infrared Photochemistry.
The efficiency of the photopolymerization technology significantly decreases when the color of materials blackens, which is contributed by the limitations of light penetration. Herein, we demonstrate rapid generation of black 3D objects up to the centimeter level in size based on melanin using near-infrared (NIR) photochemistry. Melanin, of a low absorption coefficient in the NIR range, allows thorough penetration of the 980 nm light to induce emission from upconversion nanoparticles (UCNPs) for initiating UCNP-assisted photopolymerization (UCAP). A model that describes light-attenuation gradients and dose-dependent kinetics in UCAP-guided NIR photochemistry is developed. Notably, the established model for the UCAP concept provides sufficient vertical light penetration to form scale-predictable black materials and instructs 3D printing applications. The critical control parameters were evaluated, and it was shown that complex macroscale black objects can be processed within dozens of minutes. The modeling methodologies integrated with rich functional fillers will further extend the versatility of UCAP technology in device design and manufacturing.


Globalisation as we enter the 21st century: reflections and directions for nursing education, science, research and clinical practice.
The events of September 11th, 2001 in the United States and the Bali bombings of October 2002 are chastening examples of the entangled web of the religious, political, health, cultural and economic forces we experience living in a global community. To view these forces as independent, singular, linearly deterministic entities of globalisation is irrational and illogical. Understanding the concept of globalisation has significant implications not only for world health and international politics, but also the health of individuals. Depending on an individual's political stance and world-view, globalisation may be perceived as an emancipatory force, having the potential to bridge the chasm between rich and poor or, in stark contrast, the very essence of the divide. It is important that nurses appreciate that globalisation does not pertain solely to the realms of economic theory and world politics, but also that it impacts on our daily nursing practice and the welfare of our patients. Globalisation and the closer interactions of human activity that result, have implications for international governance, policy and theory development as well as nursing education, research and clinical practice. Nurses, individually and collectively, have the political power and social consciousness to influence the forces of globalisation to improve health for all. This paper defines and discusses globalisation in today's world and its implications for contemporary nursing education, science, research and clinical practice.


On-line measurement of exhaled [11C]CO2 during PET.
The purpose of this study was to develop and evaluate a system for the continuous on-line measurement of expired 11CO2 during 11C PET studies. A detector system was developed that allowed continuous sampling of expired air during PET. Healthy volunteers (n = 4) underwent PET with [11C]CO2 during which expired air, tomographic tissue activity, and blood data were collected. The measured expired-air 11CO2 radioactivity time courses were filtered, and an envelope was extracted and compared with the time course of 11CO2/H11CO3 in blood. Good agreement was found between the shapes of the expired-air envelope and the time course in blood, enabling quantitative calibration against discrete blood samples. A system for the continuous monitoring of expired radioactivity during PET has been developed and evaluated with [11C]CO2. This monitoring enables the quantitative continuous measurement of 11CO2/H11CO3- in blood.


Microdensitometric assay of enzymatic activities in parthenogenetically activated and in vitro fertilized bovine oocytes.
To examine the paternal genome's role in reprogramming metabolic activity in one-cell embryos, we investigated metabolic aspects of bovine oocytes after in vitro maturation and in vitro fertilization and after in vitro parthenogenetic activation with a Ca2+ ionophore and 6-dimethylaminopurine. We assayed succinate dehydrogenase, lactate dehydrogenase and glucose-6-phosphate dehydrogenase activities by microspectrophotometry in immature oocytes and oocytes after maturation, in vitro fertilization and parthenogenetic activation. Succinate dehydrogenase activity significantly increased after in vitro maturation, significantly decreased after Ca2+ ionophore activation and further decreased after 6-dimethylaminopurine treatment. Lactate dehydrogenase activity showed a significant decrease in bovine oocytes after in vitro maturation, remained unchanged in Ca2+ ionophore-treated oocytes and rose significantly after 6-dimethylaminopurine treatment. This activity was dramatically reduced after in vitro fertilization, reaching absorbance levels that were not different from those in mature and Ca2+ ionophore-treated oocytes. Glucose-6-phosphate dehydrogenase activity was significantly lower in matured oocytes as compared to immature oocytes, was significantly higher after artificial activation with Ca2+ ionophore and remained constant after 6-dimethylaminopurine treatment or after in vitro fertilization. We suggest that metabolic changes involved in parthenogenetic activation are similar to those occurring after fertilization.


Unintended Consequence: Diversity as a Casualty of Eliminating United States Medical Licensing Examination Step 1 Scores.
The purpose of this study was to use a discrete-choice experiment to model the trade-offs evaluators make between academic attributes and demographics when the United States Medical Licensing Examination (USMLE) Step 1 switches to pass/fail. A discrete-choice experiment was administered to faculty members from a geographically diverse mix of 14 academic and community radiology departments in the United States from August through November 2020. Reviewers reviewed 10 applicant pairs with numeric Step 1 scores (part 1) and 10 applicant pairs with a pass Step 1 result (part 2). Applicant attributes included medical school rank, gender, race/ethnicity, USMLE Step 1 score, USMLE Step 2 score, class rank, clerkship honors, and publications. Conditional logistic regression modeled the influence of attribute levels. Two hundred twelve evaluators completed the study (response rate 59%). The most influential attribute was Step 1 score in part 1 and medical school rank in part 2. The relative importance of race/ethnicity and gender decreased by 25% and 29%, respectively, when Step 1 switches to pass/fail. Evaluators weigh race/ethnicity the strongest when applicants have the same Step 1 score (preference weights of 0.85 for African American, 1.42 for Hispanic, and 0 for White and Asian applicants). Race/ethnicity is relatively more important when Step 1 scores are higher (preference weights of 1.58 for African American, 0.90 for Hispanic, and 0 for White and Asian applicants). The loss of numeric Step 1 scores reduced the residency evaluator preference for diversity. Reviewers prioritize underrepresented-in-medicine applicants when Step 1 scores are higher and comparable with White and Asian applicants.


Prevalence of Salmonella enterica serovar Typhi infection, its associated factors and antimicrobial susceptibility patterns among febrile patients at Adare general hospital, Hawassa, southern Ethiopia.
Salmonellas enterica serovar Typhi (S.typhi) causes typhoid fever and is a global health problem, especially in developing countries like Ethiopia. But there is a little information about prevalence and factors association with S.typhi and its antimicrobial susceptibility pattern in Ethiopia especially in the study area. The aim of this study was to determine the prevalence of S.typhi infection, its associated factors and antimicrobial susceptibility pattern among patient with a febrile illness at Adare General Hospital, Hawassa, Southern Ethiopia. Hospital based cross sectional study was conducted among 422 febrile patients from May 23, 2018 to October 20, 2018. A 5 ml venous blood was collected from each febrile patient. Culture and biochemical test were performed for each isolate. Antimicrobial susceptibility testing was performed for each isolate using modified Kirby-Bauer disk diffusion techniques. In this study, the prevalence of S.typhi among febrile illness patients at Adare General Hospital was 1.6% [95% confidence interval (CI): 0.5-2.9]. The age of the study subjects were ranged from 15 to 65 years (mean age 32 years). It was observed that participants who came from rural area had 8 times (AOR 8.27: 95% CI: 1.33, 51.55) more likely to had S. typhi infection when compared with urban dwellers. The microbial susceptibility testing revealed that all six of S.typhi isolates showed sensitive to Ceftriaxone and all 6 isolates showed resistant to nalidixic acid and Cefotaxime and 5(83.3%) susceptible to Chloramphenicol and Ciprofloxaciline. Multidrug resistance (resistance to three or more antibiotics) was observed among most of the isolates. S. typhi bacteraemia is an uncommon but important cause of febrile illness in our study population. Ceftriaxone therapy is a suitable empirical antibiotic for those that are unwell and suspected of having this illness. Further surveillance is required to monitor possible hanging antibiotic resistant patterns in Ethiopia.


Variant-specific changes in persistent or resurgent sodium current in SCN8A-related epilepsy patient-derived neurons.
Missense variants in the SCN8A voltage-gated sodium channel gene are linked to early-infantile epileptic encephalopathy type 13, also known as SCN8A-related epilepsy. These patients exhibit a wide spectrum of intractable seizure types, severe developmental delay, movement disorders, and elevated risk of sudden unexpected death in epilepsy. The mechanisms by which SCN8A variants lead to epilepsy are poorly understood, although heterologous expression systems and mouse models have demonstrated altered sodium current properties. To investigate these mechanisms using a patient-specific model, we generated induced pluripotent stem cells from three patients with missense variants in SCN8A: p.R1872>L (Patient 1); p.V1592>L (Patient 2); and p.N1759>S (Patient 3). Using small molecule differentiation into excitatory neurons, induced pluripotent stem cell-derived neurons from all three patients displayed altered sodium currents. Patients 1 and 2 had elevated persistent current, while Patient 3 had increased resurgent current compared to controls. Neurons from all three patients displayed shorter axon initial segment lengths compared to controls. Further analyses focused on one of the patients with increased persistent sodium current (Patient 1) and the patient with increased resurgent current (Patient 3). Excitatory cortical neurons from both patients had prolonged action potential repolarization. Using doxycycline-inducible expression of the neuronal transcription factors neurogenin 1 and 2 to synchronize differentiation of induced excitatory cortical-like neurons, we investigated network activity and response to pharmacotherapies. Both small molecule differentiated and induced patient neurons displayed similar abnormalities in action potential repolarization. Patient induced neurons showed increased burstiness that was sensitive to phenytoin, currently a standard treatment for SCN8A-related epilepsy patients, or riluzole, an FDA-approved drug used in amyotrophic lateral sclerosis and known to block persistent and resurgent sodium currents, at pharmacologically relevant concentrations. Patch-clamp recordings showed that riluzole suppressed spontaneous firing and increased the action potential firing threshold of patient-derived neurons to more depolarized potentials. Two of the patients in this study were prescribed riluzole off-label. Patient 1 had a 50% reduction in seizure frequency. Patient 3 experienced an immediate and dramatic seizure reduction with months of seizure freedom. An additional patient with a SCN8A variant in domain IV of Nav1.6 (p.V1757>I) had a dramatic reduction in seizure frequency for several months after starting riluzole treatment, but then seizures recurred. Our results indicate that patient-specific neurons are useful for modelling SCN8A-related epilepsy and demonstrate SCN8A variant-specific mechanisms. Moreover, these findings suggest that patient-specific neuronal disease modelling offers a useful platform for discovering precision epilepsy therapies.


Correlated evolution of male and female morphologles in water striders.
Sexually antagonistic coevolution may be an important force in the evolution of sexual dimorphism. We undertake a comparative study of correlated evolution of male and female morphologies in a clade of 15 water strider species in the genus Gerris (Heteroptera: Gerridae). Earlier studies have shown that superfluous matings impose costs on females, including increased energetic expenditure and predation risk, and females therefore resist males with premating struggles. Males of some species possess grasping structures and females of some species exhibit distinct antigrasping structures, which are used to further the interests of each sex during these premating struggles. We use this understanding, combined with coevolutionary theory, to derive a series of a priori predictions concerning both the types of traits in the two sexes that are expected to coevolve and the coevolutionary dynamics of these traits expected under sexually antagonistic coevolution. We then assess the actual pattern of correlated evolution in this clade with new morphometric methods combined with standard comparative techniques. The results were in agreement with the a priori predictions. The level of armament (different abdominal structures in the two sexes) was closely correlated between the sexes across species. Males are well adapted to grasping females in species in which females are well adapted to thwart harassing males and vice versa. Furthermore, our comparative analyses supports the prediction that correlated evolution of armament in the two sexes should be both rapid and bidirectional.


Pneumonia and Streptococcus pneumoniae vaccine.
Pneumonia is an inflammatory disease of the lung, responsible for high morbidity and mortality worldwide. It is caused by bacteria, viruses, fungi, or other microorganisms. Streptococcus pneumoniae, a gram-positive bacterium with over 90 serotypes, is the most common causative agent. Moreover, comorbid factors including heart failure, renal disease, and pulmonary disease could increase the risk of pneumococcal pneumonia. Since the advent of the pneumococcal vaccine in the 1980s, the incidence of pneumonia has decreased significantly. However, current vaccines confer only limited protection against serotypes included in the vaccine. Thus, to overcome this limitation, new types of pneumococcal vaccines have been sought and under clinical trials. In this review, we discuss pneumonia and summarize the various types of pneumococcal vaccines in progress.


Enantioselective synthesis of 3,4-dihydro-1,2-oxazepin-5(2H)-ones and 2,3-dihydropyridin-4(1H)-ones from β-substituted β-hydroxyaminoaldehydes.
The synthesis of 3,4-dihydro-1,2-oxazepin-5(2H)-ones and 2,3-dihydropyridin-4(1H)-ones from β-substituted β-hydroxyaminoaldehydes is reported. The β-hydroxyaminoaldehydes were prepared by enantioselective organocatalytic 1,4-addition of N-tert-butyl (tert-butyldimethylsilyl)oxycarbamate to α,β-unsaturated aldehydes (MacMillan protocol). Alkyne addition to the aldehydes followed by alcohol oxidation furnished N-Boc O-TBS-protected β-aminoynones. Removal of the TBS protecting group initiated a 7-endo-dig cyclization to yield previously unknown 3,4-dihydro-1,2-oxazepin-5(2H)-ones. Reductive cleavage of the N-O bond of the oxazepinones and Boc-deprotection provided 2-substituted 2,3-dihydropyridin-4(1H)-ones via 6-endo-trig cyclization. 2,3-Dihydropyridin-4(1H)-ones are versatile intermediates that have been used for the synthesis of many alkaloids. The new protocol allows the synthesis of 3-dihydropyridin-4(1H)-ones carrying an array of substituents at C2 that cannot be prepared from commercial β-amino acids or by one-carbon homologation of proteinogenic amino acids. The use of readily available β-hydroxylaminoaldehydes expands the utility of our previously reported method to prepare 2,3-dihydropyridin-4(1H)-ones from β-amino acids as the source of diversity and chirality. A broad substrate scope is possible because β-aminoaldehydes can be prepared from α,β-unsaturated aldehydes by an enantioselective organocatalytic process.


Partitioning of selected antioxidants in mayonnaise.
This study examined partitioning of alpha-, beta-, and gamma-tocopherol and six polar antioxidants (Trolox, ferulic acid, caffeic acid, propyl gallate, gallic acid, and catechin) in mayonnaise. Partitioning of antioxidants between different phases was determined after separation of mayonnaise by either (a) centrifugation + ultracentrifugation or (b) centrifugation + dialysis. Antioxidants partitioned in accordance with their chemical structure and polarity: Tocopherols were concentrated in the oil phase (93-96%), while the proportion of polar antioxidants in the oil phase ranged from 0% (gallic acid and catechin) to 83% (Trolox). Accordingly, proportions of 6% (Trolox) to 80% (gallic acid and catechin) were found in the aqueous phase. Similar trends were observed after dialysis. After ultracentrifugation, large proportions of polar antioxidants were found in the "emulsion phase" and the "precipitate" (7-34% and 2-7%, respectively). This indicated entrapment of antioxidants at the oil-water interface in mayonnaise. The results signify that antioxidants partitioning into different phases of real food emulsions may vary widely.


Dendritic cells are crucial for cardiovascular remodeling and modulate neutrophil gelatinase-associated lipocalin expression upon mineralocorticoid receptor activation.
Adaptive immunity is crucial in cardiovascular and renal inflammation/fibrosis upon hyperactivation of mineralocorticoid receptor. We have previously demonstrated that dendritic cells can respond to mineralocorticoid receptor activation, and the neutrophil gelatinase-associated lipocalin (NGAL) in dendritic cells is highly increased during aldosterone (Aldo)/mineralocorticoid receptor-dependent cardiovascular damage. However, the interrelationship among dendritic cells, target organs inflammation/fibrosis induced by mineralocorticoid receptor, and NGAL-dependence remains unknown. We studied the role of dendritic cells in mineralocorticoid receptor-dependent tissue remodeling and whether NGAL can modulate the inflammatory response of dendritic cells after mineralocorticoid receptor activation. Cardiovascular and renal remodeling induced by Aldo and high-salt diet [nephrectomy-Aldo-salt (NAS) model] were analyzed in CD11c.DOG mice, a model which allows dendritic cells ablation by using diphtheria toxin. In addition, in-vitro studies in NGAL-knock out dendritic cells were performed to determine the immunomodulatory role of NGAL upon Aldo treatment. The ablation of dendritic cells prevented the development of cardiac hypertrophy, perivascular fibrosis, and the overexpression of NGAL, brain natriuretic peptide, and two profibrotic factors induced by NAS: collagen 1A1 and connective tissue growth factor. We determined that dendritic cells were not required to prevent renal hypertrophy/fibrosis induced by NAS. Between different immune cells analyzed, we observed that NGAL abundance was higher in antigen-presenting cells, while in-vitro studies showed that mineralocorticoid receptor stimulation in dendritic cells favored NGAL and IL-23 expression (p19 and p40 subunits), which are involved in the development of fibrosis and the Th17-driven response, respectively. NGAL produced by dendritic cells may play a pivotal role in the activation of adaptive immunity that leads to cardiovascular fibrosis during mineralocorticoids excess.


The purification of poly(a)-containing RNA by affinity chromatography.
The vast majority of eukaryotic mRNA molecules contain tracts of poly(adenylic) acid, up to 250 bases in length, at the 3' end. This property is very useful from the point of view of mRNA extraction because it forms the basis of a convenient and simple affinity chromatography procedure (1). Under high salt conditions (0.3-0.5M NaCl or KCl), poly(A) will hybridize to oligo(dT)-cellulose or poly(U)-Sepharose. These commercially available materials consist of polymers of about 10-20 nucleotides, covalently bound to a carbohydrate support, and bind RNA containing a poly(A) tract as short as 20 residues. Ribosomal and transfer RNAs do not possess poly(A) sequences and will not bind (see Note 1).


Pineal gland volume in primary insomnia and healthy controls: a magnetic resonance imaging study.
Little is known about the relation between pineal volume and insomnia. Melatonin promotes sleep processes and, administered as a drug, it is suitable to improve primary and secondary sleep disorders in humans. Recent magnetic resonance imaging studies suggest that human plasma and saliva melatonin levels are partially determined by the pineal gland volume. This study compares the pineal volume in a group of patients with primary insomnia to a group of healthy people without sleep disturbance. Pineal gland volume (PGV) was measured on the basis of high-resolution 3 Tesla MRI (T1-magnetization prepared rapid gradient echo) in 23 patients and 27 controls, matched for age, gender and educational status. Volume measurements were performed conventionally by manual delineation of the pineal borders in multi-planar reconstructed images. Pineal gland volume was significantly smaller (P < 0.001) in patients (48.9 ± 26.6 mm(3) ) than in controls (79 ± 30.2 mm(3) ). In patients PGV correlated negatively with age (r = -0.532; P = 0.026). Adjusting for the effect of age, PGV and rapid eye movement (REM) latency showed a significant positive correlation (rS = 0.711, P < 0.001) in patients. Pineal volume appears to be reduced in patients with primary insomnia compared to healthy controls. Further studies are needed to clarify whether low pineal volume is the basis or the consequence of functional sleep changes to elucidate the molecular pathology for the pineal volume loss in primary insomnia.


Paleolithic and Mediterranean Diet Pattern Scores Are Inversely Associated with Biomarkers of Inflammation and Oxidative Balance in Adults.
Chronic inflammation and oxidative balance are associated with poor diet quality and risk of cancer and other chronic diseases. A diet-inflammation/oxidative balance association may relate to evolutionary discordance. We investigated associations between 2 diet pattern scores, the Paleolithic and the Mediterranean, and circulating concentrations of 2 related biomarkers, high-sensitivity C-reactive protein (hsCRP), an acute inflammatory protein, and F2-isoprostane, a reliable marker of in vivo lipid peroxidation. In a pooled cross-sectional study of 30- to 74-y-old men and women in an elective outpatient colonoscopy population (n = 646), we created diet scores from responses on Willett food-frequency questionnaires and measured plasma hsCRP and F2-isoprostane concentrations by ELISA and gas chromatography-mass spectrometry, respectively. Both diet scores were calculated and categorized into quintiles, and their associations with biomarker concentrations were estimated with the use of general linear models to calculate and compare adjusted geometric means, and via unconditional ordinal logistic regression. There were statistically significant trends for decreasing geometric mean plasma hsCRP and F2-isoprostane concentrations with increasing quintiles of the Paleolithic and Mediterranean diet scores. The multivariable-adjusted ORs comparing those in the highest with those in the lowest quintiles of the Paleolithic and Mediterranean diet scores were 0.61 (95% CI: 0.36, 1.05; P-trend = 0.06) and 0.71 (95% CI: 0.42, 1.20; P-trend = 0.01), respectively, for a higher hsCRP concentration, and 0.51 (95% CI: 0.27, 0.95; P-trend 0.01) and 0.39 (95% CI: 0.21, 0.73; P-trend = 0.01), respectively, for a higher F2-isoprostane concentration. These findings suggest that diets that are more Paleolithic- or Mediterranean-like may be associated with lower levels of systemic inflammation and oxidative stress in humans.


Coherent Superpositions of Photon Creation Operations and Their Application to Multimode States of Light.
We present a concise review of recent experimental results concerning the conditional implementation of coherent superpositions of single-photon additions onto distinct field modes. Such a basic operation is seen to give rise to a wealth of interesting and useful effects, from the generation of a tunable degree of entanglement to the birth of peculiar correlations in the photon numbers and the quadratures of multimode, multiphoton, states of light. The experimental investigation of these properties will have an impact both on fundamental studies concerning, for example, the quantumness and entanglement of macroscopic states, and for possible applications in the realm of quantum-enhanced technologies.


The orthopaedic error index: development and application of a novel national indicator for assessing the relative safety of hospital care using a cross-sectional approach.
The Orthopaedic Error Index for hospitals aims to provide the first national assessment of the relative safety of provision of orthopaedic surgery. Cross-sectional study (retrospective analysis of records in a database). The National Reporting and Learning System is the largest national repository of patient-safety incidents in the world with over eight million error reports. It offers a unique opportunity to develop novel approaches to enhancing patient safety, including investigating the relative safety of different healthcare providers and specialties. We extracted all orthopaedic error reports from the system over 1 year (2009-2010). The Orthopaedic Error Index was calculated as a sum of the error propensity and severity. All relevant hospitals offering orthopaedic surgery in England were then ranked by this metric to identify possible outliers that warrant further attention. 155 hospitals reported 48 971 orthopaedic-related patient-safety incidents. The mean Orthopaedic Error Index was 7.09/year (SD 2.72); five hospitals were identified as outliers. Three of these units were specialist tertiary hospitals carrying out complex surgery; the remaining two outlier hospitals had unusually high Orthopaedic Error Indexes: mean 14.46 (SD 0.29) and 15.29 (SD 0.51), respectively. The Orthopaedic Error Index has enabled identification of hospitals that may be putting patients at disproportionate risk of orthopaedic-related iatrogenic harm and which therefore warrant further investigation. It provides the prototype of a summary index of harm to enable surveillance of unsafe care over time across institutions. Further validation and scrutiny of the method will be required to assess its potential to be extended to other hospital specialties in the UK and also internationally to other health systems that have comparable national databases of patient-safety incidents.


[The forms of the intercellular contacts of Leishmania in culture].
Two kinds of intercellular interactions have been observed in cultures of 12 investigated Leishmania species (L. major, L. tropica, L. donovani, L. infantum, L. sp. ZMA, L. mexicana, L. hertigi, L. braziliensis, L. tarentolae, L. adleri, L. gymnodactyli, L. gulickae). The first kind looks as adhesion of two specimens with their fore-ends. This way is characteristic of promastigotes of different morphotypes as well as of the interphase and dividing organisms to be most frequently seen in L. mexicana and L. gymnodactyli, and in both dark and lucid forms. The second kind of intercellular interactions involves a coupled adhesion of morphologically similar promastigotes with free ends of flagella. It is most characteristic of L. gymnodactyli and specially of dark promastigotes. Proves are provided that both the kinds of cell interactions are not associated with cell division, that they may only partially be connected with the phenomenon of rosette formation, and that they represent different phenomena. It is supposed that the intercellular contacts with the fore-ends may reflect gene exchanges in two partners, with a possible involvement of the kinetoplast DNA.


On the genetic basis of face cognition and its relation to fluid cognitive abilities.
The oxytocin and the dopaminergic systems have turned out to be highly relevant for social abilities and cognition. Therefore, we examined the association between two functional gene polymorphisms and face cognition (FC) in a multivariate study (N = 250) by applying structural equation modeling. The catechol-O-methyltransferase (COMT) val158met polymorphism influences the enzyme activity of COMT, which affects the prefrontal dopamine concentration. The rs226849 is a single-nucleotide polymorphism located in the promoter region of the oxytocin receptor (OXTR) gene, modulating the mRNA expression. By modeling a general fluid ability factor (defined by working memory and reasoning) and nested FC factors, we tested genetic contributions to FC, after controlling for variance in FC that was also associated with fluid abilities. In line with several previous studies, we found a significant association between the COMT genotype and fluid abilities (Gf) but not with FC. The association between the oxytocin polymorphism and Gf was opposite in direction for men and women. Women with the C(+) genotype performed better on Gf tasks than those with the C(-) genotype. Conversely, men with the C(-) genotype performed better than those with the C(+) genotype. There was no significant association between OXTR and the nested FC factor. Therefore, the relationship between the oxytocin polymorphism and FC can be fully accounted for by Gf. The sex specificity of this relationship is a novel finding and warrants a mechanistic explanation.


Towards a mathematical framework to inform neural network modelling via polynomial regression.
Even when neural networks are widely used in a large number of applications, they are still considered as black boxes and present some difficulties for dimensioning or evaluating their prediction error. This has led to an increasing interest in the overlapping area between neural networks and more traditional statistical methods, which can help overcome those problems. In this article, a mathematical framework relating neural networks and polynomial regression is explored by building an explicit expression for the coefficients of a polynomial regression from the weights of a given neural network, using a Taylor expansion approach. This is achieved for single hidden layer neural networks in regression problems. The validity of the proposed method depends on different factors like the distribution of the synaptic potentials or the chosen activation function. The performance of this method is empirically tested via simulation of synthetic data generated from polynomials to train neural networks with different structures and hyperparameters, showing that almost identical predictions can be obtained when certain conditions are met. Lastly, when learning from polynomial generated data, the proposed method produces polynomials that approximate correctly the data locally.


The detection of trans gene fragments of hEPO in gene doping model mice by Taqman qPCR assay.
With the rapid progress of genetic engineering and gene therapy methods, the World Anti-Doping Agency has raised concerns regarding gene doping, which is prohibited in sports. However, there is no standard method available for detecting transgenes delivered by injection of naked plasmids. Here, we developed a detection method for detecting transgenes delivered by injection of naked plasmids in a mouse model that mimics gene doping. Whole blood from the tail tip and one piece of stool were used as pre-samples of injection. Next, a plasmid vector containing the human erythropoietin (hEPO) gene was injected into mice through intravenous (IV), intraperitoneal (IP), or local muscular (IM) injection. At 1, 2, 3, 6, 12, 24, and 48 h after injection, approximately 50 µL whole blood was collected from the tail tip. One piece of stool was collected at 6, 12, 24, and 48 h. From each sample, total DNA was extracted and transgene fragments were analyzed by Taqman quantitative PCR (qPCR) and SYBR green qPCR. In whole blood DNA samples evaluated by Taqman qPCR, the transgene fragments were detected at all time points in the IP sample and at 1, 2, 3, 6, and 12 h in the IV and IM samples. In the stool-DNA samples, the transgene fragments were detected at 6, 12, 24, and 48 h in the IV and IM samples by Taqman qPCR. In the analysis by SYBR green qPCR, the transgene fragments were detected at some time point in both specimens; however, many non-specific amplicons were detected. These results indicate that transgene fragments evaluated after each injection method of naked plasmids were detected in whole-blood and stool DNA samples. These findings may facilitate the development of methods for detecting gene doping.


Thermodynamics and docking of agonists to the β(2)-adrenoceptor determined using [(3)H](R,R')-4-methoxyfenoterol as the marker ligand.
G protein-coupled receptors (GPCRs) are integral membrane proteins that change conformation after ligand binding so that they can transduce signals from an extracellular ligand to a variety of intracellular components. The detailed interaction of a molecule with a G protein-coupled receptor is a complicated process that is influenced by the receptor conformation, thermodynamics, and ligand conformation and stereoisomeric configuration. To better understand the molecular interactions of fenoterol analogs with the β(2)-adrenergic receptor, we developed a new agonist radioligand for binding assays. [(3)H](R,R')-methoxyfenoterol was used to probe the binding affinity for a series of fenoterol stereoisomers and derivatives. The results suggest that the radioligand binds with high affinity to an agonist conformation of the receptor, which represents approximately 25% of the total β(2)-adrenoceptor (AR) population as determined with the antagonist [(3)H]CGP-12177. The β(2)-AR agonists tested in this study have considerably higher affinity for the agonist conformation of the receptor, and K(i) values determined for fenoterol analogs model much better the cAMP activity of the β(2)-AR elicited by these ligands. The thermodynamics of binding are also different when interacting with an agonist conformation, being purely entropy-driven for each fenoterol isomer, rather than a mixture of entropy and enthalpy when the fenoterol isomers binding was determined using [(3)H]CGP-12177. Finally, computational modeling identified the molecular interactions involved in agonist binding and allow for the prediction of additional novel β(2)-AR agonists. The study underlines the possibility of using defined radioligand structure to probe a specific conformation of such shape-shifting system as the β(2)-adrenoceptor.