High peak-power mid-infrared ZnGeP₂ optical parametric oscillator pumped by a Tm:fiber master oscillator power amplifier system.
We report on the utilization of a novel Tm:fiber laser source for mid-IR ZnGeP2 (ZGP) optical parametric oscillator (OPO) pumping. The pump laser is built in a master oscillator power-amplifier configuration delivering up to 3.36 W of polarized, diffraction limited output power with 7 ns pulse duration and 4 kHz repetition rate. This corresponds to a peak power of ∼121 kW and a pulse energy of ∼0.84 mJ. With this source, we generated 27.9 kW of total mid-IR peak power in a doubly resonant oscillator (DRO) configuration. This is, to the best of our knowledge, the highest ever demonstrated mid-IR peak power from a directly Tm:fiber laser pumped ZGP OPO. Moreover, a DRO output with about 284 μJ of total mid-IR pulse energy was demonstrated using 100 ns pump pulses. The wavelength tuning of the idler was extended to 6 μm with lower output power in another OPO experiment.


Shorter telomere length predicts poorer immunological recovery in virologically suppressed HIV-1-infected patients treated with combined antiretroviral therapy.
Successful combined antiretroviral therapy (cART) does not always result in complete CD4 T-cell recovery despite the effective control of HIV replication. Because telomere dysregulation can lead to an abnormal cell proliferation, we hypothesized that the lack of CD4 recovery may be related to telomere defects; We thus evaluated the association between telomere length (TL) and CD4 T-cell recovery 48 weeks after cART initiation in virologically suppressed patients, and its possible relationship to oxidative stress (OS) and nitrosative stress (NOx) markers. We studied HIV-infected patients on stable cART who achieved a viral load <50 copies per milliliter after 48 weeks of their first cART. Leukocyte TL was measured and categorized into tertiles. We calculated mean increases in CD4 T-cell at 48 weeks from cART initiation and used multivariate linear regression models to estimate differences in mean increases according to tertiles of TL. One hundred thirty-two patients, 86% male, 81% <50 years at cART initiation were studied. Mean increases in CD4 were greater in patients with long TL than in those with medium and short TLs (P = 0.007). After adjustment for sex, age, CD4 T-cell counts, viral load, and hepatitis C infection at cART initiation, differences in mean CD4 T-cell count increases according to TL remained statistically significant (P = 0.02). Additional adjustment for NOx and OS did not change the results. A lower immunological response despite a successful virological response is associated with a shorter TL. The effect is not related to NOx or OS.


The four and a half LIM-only protein 2 (FHL2) activates transforming growth factor β (TGF-β) signaling by regulating ubiquitination of the E3 ligase Arkadia.
Arkadia is a RING-based ubiquitin ligase that positively regulates TGF-β signaling by targeting several pathway components for ubiquitination and degradation. However, little is known about the mechanisms controlling Arkadia activity. Here we show that the LIM-only protein FHL2 binds and synergistically cooperates with Arkadia to activate Smad3/Smad4-dependent transcription. Knockdown of FHL2 by RNA interference decreases Arkadia level and restricts the amplitude of Arkadia-induced TGF-β target gene responses. We found that Arkadia is ubiquitinated via K63- and K27-linked polyubiquitination. A single mutation at the RING domain that abolishes the E3 activity diminishes Arkadia ubiquitination, indicating that this modification partly involves autocatalytic process. Mutation of seven lysines at the C-terminal region of Arkadia severely impairs ubiquitination through the K27 but not the K63 linkage and slows down the turnover of Arkadia, suggesting that K27-linked polyubiquitination might promote proteolysis-dependent regulation of Arkadia. We show that FHL2 increases the half-life of Arkadia through inhibition of ubiquitin chain assembly on the protein, which provides a molecular basis for functional cooperation between Arkadia and FHL2 in enhancing TGF-β signaling. Our study uncovers a novel regulatory mechanism of Arkadia by ubiquitination and identifies FHL2 as important regulator of Arkadia ubiquitination and TGF-β signal transduction.


Clinical psychology and disability studies: bridging the disciplinary divide on mental health and disability.
Clinical psychology and disability studies have traditionally occupied very different academic, philosophical and political spaces. However, this paper aims to illustrate the positive consequences and implications of attempts to understand and bridge this disciplinary divide. A narrative review format was used with evidence selected pragmatically as opposed to systematically. The construction of the argument determined the evidence selected. The concept of psycho-emotional disablism, which originated within disability studies, is argued to be a useful concept to bridge the divide between understandings of distress from both disability studies and clinical psychology perspectives. However, this can be usefully augmented by psychological research on the mechanisms through which disablism can affect individuals. Perspectives from both disability studies and clinical psychology can be usefully combined to bring important new perspectives; combined, these perspectives should help - on theoretical, service and social levels - to improve the mental health of disabled people. Implications for Rehabilitation Mental health is an important determinant of overall health-related quality of life and psychological therapy should be available for those disabled people who would value it. Psychological therapists working with disabled people should be more aware of the challenging social context in which disabled people live. Understandings of distress should not just include individual factors but also incorporate the psychological impact of stresses caused by societal barriers preventing inclusion. Psychologists should be more willing to work and engage at a societal and political level to influence change.


Survey design for precise fire management conservation targets.
Common goals of ecological fire management are to sustain biodiversity and minimize extinction risk. A novel approach to achieving these goals determines the relative proportions of vegetation growth stages (equivalent to successional stages, which are categorical representations of time since fire) that maximize a biodiversity index. The method combines data describing species abundances in each growth stage with numerical optimization to define an optimal growth-stage structure that provides a conservation-based operational target for managers. However, conservation targets derived from growth-stage optimization are likely to depend critically on choices regarding input data. There is growing interest in the use of growth-stage optimization as a basis for fire management, thus understanding of how input data influence the outputs is crucial. Simulated data sets provide a flexible platform for systematically varying aspects of survey design and species inclusions. We used artificial data with known properties, and a case-study data set from southeastern Australia, to examine the influence of (1) survey design (total number of sites and their distribution among growth stages) and (2) species inclusions (total number of species and their level of specialization) on the precision of conservation targets. Based on our findings, we recommend that survey designs for precise estimates would ideally involve at least 80 sites, and include at least 80 species. Greater numbers of sites and species will yield increasingly reliable results, but fewer might be sufficient in some circumstances. An even distribution of sites among growth stages was less important than the total number of sites, and omission of species is unlikely to have a major influence on results as long as several species specialize on each growth stage. We highlight the importance of examining the responses of individual species to growth stage before feeding survey data into the growth-stage optimization black box, and advocate use of a resampling procedure to determine the precision of results. Collectively, our findings form a reproducible guide to designing ecological surveys that yield precise conservation targets through growth-stage optimization, and ultimately help sustain biodiversity in fire-prone systems.


Associations between Dietary Patterns and Post-Bronchodilation Lung Function in the SAPALDIA Cohort.
Chronic obstructive pulmonary disease (COPD) is not restricted to smokers. Dietary habits may contribute to the disease occurrence. Epidemiological studies point to a protective effect of fruit and vegetable intake against COPD. To investigate the associations between dietary patterns and parameters of lung function related to COPD in the Swiss Cohort Study on Air Pollution and Lung and Heart Diseases in Adults (SAPALDIA). Data were included from the second follow-up assessment of the SAPALDIA cohort in 2010-2011 using a food frequency questionnaire. Principal component factor analysis was used to derive dietary patterns, whose association with FEV1, FEV1/FVC, FEF2575, and COPD was investigated by applying multivariate regression analyses. After adjustment for potential confounders, the "prudent dietary pattern" characterised by the predominant food groups vegetables, fruits, water, tea and coffee, fish, and nuts was positively associated with FEV1 (increase of 40 mL per SD, p < 0.001). Also for factor 3 ("high-carbohydrate diet"), we found a significant positive association with FEV1 (with an increase per SD of 36 mL, p = 0.006). The main results are consistent with a protective effect of a diet rich in fruits, vegetables, fish, and nuts against age-related chronic respiratory disease. If confirmed in prospective cohorts, our results may guide nutritional counselling towards respiratory health promotion.


The 10 commandments of nutrition: 2014.
The US Special Operations Command requires sound recommendations on nutrition to ensure optimal performance of Special Operations personnel. New information continues to emerge, and previous recommendations need to be modified as the evidence base continues to grow. The first 10 Commandments of Nutrition were published in the SEAL professional journal Full Mission Profile in 1992, published for the second time in this journal in 2005, and now revised a second time to reflect the newest science. Whether you are part of the Special Operations Forces (SOF) community or an athlete seeking to improve your performance, these are simple and helpful nutrition guidelines to follow.


Visual Outcomes and Higher Order Aberrations Following LASIK on Eyes with Low Myopia and Astigmatism.
Laser-Assisted in situ Keratomileusis (LASIK) can induce corneal aberrations that can impact vision and patient satisfaction. Recent developments in laser technologies have helped minimise these aberrations. To assess the quality of vision and change in Higher-Order Aberrations (HOAs) following wavefront-optimized LASIK in low-myopic astigmatic patients. LASIK was performed on a total of 76 eyes in patients with myopia <4.0 D and cylinder <2.0 D using the WaveLight® EX500 excimer and FS200 femtosecond laser platform. Visual acuity, contrast sensitivity and HOAs were measured at 1 and 3 months postoperatively and compared to preoperative values. Subjective quality of vision was assessed pre- and postoperatively using a VF14 questionnaire. Mean postoperative Spherical Equivalent (SE) was -0.09 ± 0.26 µm with 95% of patients within ± 0.5 D of attempted SE. Postoperative uncorrected distance visual acuity was 20/20 or better for 96% of patients. Contrast sensitivity increased against horizontal and vertical gratings at all spatial frequencies except for vertical gratings at 18 cycles/degree. Spherical aberration and total HOA increased by 0.085 µm and 0.13 µm respectively. The mean VF14 score increased from 89.2 ± 16.7% to 99 ± 1.4% postoperatively. LASIK performed using the WaveLight® EX500 excimer and WaveLight® FS200 laser platform provided improved contrast sensitivity and visual acuity with minimal introduction of HOAs, making it a suitable platform for low myopic astigmatic patients.


Pressure distribution on the anatomic landmarks of the knee and the effect of kneepads.
This study examines stress transmitted to anatomic landmarks of the knee (patella, combined patella tendon and tibial tubercle) while in static kneeling postures without kneepads and while wearing two kneepads commonly worn in the mining industry. Ten subjects (7 male, 3 female) simulated postures utilized in low-seam mines: kneeling in full flexion; kneeling at 90° of knee flexion; and kneeling on one knee while in one of three kneepad states (no kneepads, non-articulated kneepads, and articulated kneepads). For each posture, peak and mean pressure on the anatomic landmarks of the knee were obtained. The majority of the pressure was found to be transmitted to the knee via the combined patellar tendon and tibial tubercle rather than through the patella. While the kneepads tested decreased the maximum pressure experienced at the combined patellar tendon and tibial tubercle, peak pressures of greater than 25 psi were still experienced over structures commonly injured in mining (e.g. bursa sac - bursitis/Miner's Knee). The major conclusion of this study is that novel kneepad designs that redistribute the stresses at the knee across a greater surface area and to other regions of the leg away from key structures of the knee are needed.


Assisted reproductive technology in Europe, 2008: results generated from European registers by ESHRE.
This 12th European IVF-monitoring (EIM) report presents the results of treatments involving assisted reproductive technology (ART) initiated in Europe during 2008. From 36 countries (3 more compared with 2007), 1051 clinics reported 532 260 treatment cycles including: IVF (124 539), ICSI (280 552), frozen embryo replacements (FER, 97 120), egg donation (ED, 13 609), in vitro maturation (IVM, 562), preimplantation genetic diagnosis/screening (PGD/PGS, 2875) and frozen oocyte replacements (FOR, 4080). Overall, this represents a 7.9% increase in the activity since 2007, which is mainly related to an increase in cycles from almost all registers and only partially to the new countries entering EIM (Estonia, Kazakhstan, Moldova and Romania, 5480 cycles in total). European data on intrauterine insemination using husband/partner's (IUI-H) and donor (IUI-D) semen were reported from 27 and 21 countries, respectively. A total of 144 509 IUI-H (+1.5%) and 24 960 IUI-D (-4.3%) cycles were included. In 19 countries where all clinics reported to the ART register, a total of 350 143 ART cycles were performed in a population of 369.8 million, corresponding to 947 cycles per million inhabitants. For IVF, the clinical pregnancy rates per aspiration and per transfer were 28.5 and 32.5%, respectively, and for ICSI the corresponding rates were 28.7 and 31.9%. In FER cycles, the pregnancy rate per thawing was 19.3%. The delivery rate after IUI was 9.1% for IUI-H and 13.8% for IUI-D. In IVF and ICSI cycles, one, two, three and four or more embryos were transferred in 22.4, 53.2, 22.3 and 2.1%, respectively. The proportions of singleton, twin and triplet deliveries after IVF and ICSI (combined) were 78.3, 20.7 and 1.0%, respectively, resulting in a total multiple delivery rate of 21.7%, compared with 22.3% in 2007, 20.8% in 2006 and 21.8% in 2005. In FER cycles, the multiple delivery rate was 13.7% (13.4% twins and 0.3% triplets). In women undergoing IUI, twin and triplet deliveries occurred in 10.6% and 0.7% with IUI-H and in 9.4 and 0.3% with IUI-D, respectively. In comparison with previous years, there was an increase in the reported number of ART cycles in Europe. For the first time in 5 years, the pregnancy rates failed to show a year-on-year increase. Compared with 2007, the number of transfers of multiple embryos (3+) and a multiple delivery rate showed a marginal decline.


Delayed sleep phase syndrome in adolescents: prevalence and correlates in a large population based study.
The aims of this study were to estimate the prevalence of Delayed Sleep Phase Syndrome (DSPS) in adolescence, and to examine the association to insomnia and school non-attendance. Data stem from a large population based study in Hordaland County in Norway conducted in 2012, the ung@hordaland study. In all, 10,220 adolescents aged 16-18 years (54% girls) provided self-reported data on a range of sleep parameters: DSPS was defined according to the International Classification of Sleep Disorders, Revised (ICSD-R) criteria, while insomnia was defined according to the Quantitative Criteria for Insomnia. Other sleep parameters included time in bed, sleep duration, sleep efficiency, oversleeping, sleep onset latency, wake after sleep onset, subjective sleep need, sleep deficiency, sleepiness and tiredness. Sleep data were calculated separately for weekdays and weekends. Data on school non-attendance were provided by official registers. The prevalence of DSPS was 3.3%, and significantly higher among girls (3.7%) than boys (2.7%). There was a strong overlap between DSPS and insomnia, with more than half of the adolescents with DSPS also meeting the criteria for insomnia (53.8% for boys and 57.1% for girls). Adolescents with DSPS had significantly higher odds ratios (OR) of non-attendance at school. After adjusting for sociodeographical factors, insomnia and depression, the adjusted ORs for days of non-attendance were OR = 3.22 (95% CI: 1.94-5.34) for boys and OR = 1.87 (95% CI: 1.25-2.80) for girls. A similar effect was found for hours of non-attendance for boys, with an adjusted OR = 3.05 (95% CI: 1.89-4.92). The effect for girls was no longer significant after full adjustment (OR =1.48 [95% CI: 0.94-2.32]). This is one of the first studies to estimate the prevalence of DSPS in adolescents. The high prevalence of DSPS, and overlap with insomnia, in combination with the odds of school non-attendance, suggest that a broad and thorough clinical approach is warranted when adolescents present with symptoms of DSPS.


Intracranial dural arteriovenous malformations: factors predisposing to an aggressive neurological course.
The natural history of cranial dural arteriovenous malformations (AVM's) is highly variable. The authors present their clinical experience with 17 dural AVM's in adults, including 10 cases with an aggressive neurological course (strictly defined as hemorrhage or progressive focal neurological deficit other than ophthalmoplegia). Two of these 10 patients died prior to surgical intervention and a third was severely disabled by intracerebral hemorrhage. Six patients underwent surgical resection of their dural AVM, with preparatory embolization in two cases. One patient received embolization and radiation therapy without surgery. Six of the seven cases without an aggressive neurological course were treated conservatively, and the seventh patient underwent embolization of a cavernous sinus dural AVM because of worsening ophthalmoplegia. In order to clarify features associated with aggressive behavior, a comprehensive meta-analysis was performed on 360 additional dural AVM's reported in the literature with sufficiently detailed clinical and angiographic information. The location and angiographic features of 100 aggressive cases were compared to those of 277 benign cases. No location of dural AVM's was immune from aggressive neurological behavior; however, an aggressive neurological course was least often associated with cases involving the transverse-sigmoid sinuses and cavernous sinus and most often associated with cases at the tentorial incisura. Contralateral contribution to arterial supply and rate of shunting (high vs. low flow) did not correlate with aggressive neurological behavior as defined. Leptomeningeal venous drainage, variceal or aneurysmal venous dilations, and galenic drainage correlated significantly (p less than 0.05) with aggressive neurological presentation. The latter three angiographic features often coexisted in the same dural AVM. It is concluded that these features significantly increase the natural risk of dural AVM's, and warrant a more vigilant therapeutic strategy.


Utilization of the Electronic Health Record to Improve Provision of Smoking Cessation Resources for Vascular Surgery Inpatients.
Identification of hospitalized patients who smoke has shown significant improvement in recent years, but provision of evidence-based tobacco cessation treatment remains a challenge. This study evaluated the utilization of an electronic health record (EHR) to facilitate implementation of evidence-based clinical practice guidelines for smoking cessation on a vascular surgery inpatient unit. A pre-and post-intervention cohort study was conducted over 6 months at a single academic medical center with a comprehensive EHR. All patients admitted to the vascular surgery service and documented as current smokers were included. A vascular surgery discharge order set with an evidence-based smoking cessation module was developed and implemented. The primary outcome was prescription of nicotine replacement therapy (NRT) at the time of discharge. The secondary outcome was referral for smoking cessation counseling at the time of discharge. There were 52 and 42 smokers in the pre-and post-intervention cohorts, respectively. Over the 3 months following implementation of the EHR order set, prescription of NRT at the time of discharge did not change significantly (27% vs 19%, p=0.30). Referral for outpatient smoking cessation counseling increased in the post-intervention group, but did not reach significance (64% vs 72%, p=0.20). Implementation of a brief tobacco dependence treatment order set in an existing EHR increased cessation counseling referrals on a vascular surgery inpatient unit. One potential limitation of the study was the modest sample size. Not being able to make smoking cessation treatment a mandatory component in discharge orders may also have contributed to the modest effect. Assessing the differential effect of EHR-based order implementation will be important in future research on this topic.


Nature of ribonucleic acid stimulated by streptomycin in the absence of protein synthesis.
Freda, Celia E. (University of Pennsylvania School of Medicine, Philadelphia), and Seymour S. Cohen. Nature of ribonucleic acid stimulated by streptomycin in the absence of protein synthesis. J. Bacteriol. 92:1680-1688. 1966.-The ribonucleic acid (RNA) synthesized in a thymineless, arginineless, uracil-less Escherichia coli strain 15 in the absence of arginine was characterized by sucrose density gradient centrifugation. About 60% of this RNA had sedimentation rates in the range between 4S and 16S, and the remainder was comprised of the 23S and 16S ribosomal components. On addition of streptomycin for 1 hr in the absence of the amino acid, there was an inhibition of synthesis of material of 4S to 16S, whereas 16S RNA was slightly stimulated. Between 1 and 3 hr after addition of the antibiotic, during the precipitous killing of the bacteria in the arginine-deficient culture, the synthesis of 16S ribosomal RNA was specifically and sharply stimulated.


DNA-mediated exciton coupling and electron transfer between donor and acceptor stilbenes separated by a variable number of base pairs.
The synthesis, steady-state spectroscopy, and transient absorption spectroscopy of DNA conjugates possessing both stilbene electron donor and electron acceptor chromophores are described. These conjugates are proposed to form nicked DNA dumbbell structures in which a stilbenedicarboxamide acceptor and stilbenediether donor are separated by variable numbers of A-T or G-C base pairs. The nick is located either adjacent to one of the chromophores or between two of the bases. Thermal dissociation profiles indicate that stable structures are formed possessing as few as two A-T base pairs. Circular dichroism (CD) spectra in the base pair region are characteristic of B-DNA duplex structures, whereas CD spectra at longer wavelengths display two bands attributed to exciton coupling between the two stilbenes. The sign and intensity of these bands are dependent upon both the distance between the chromophores and the dihedral angle between their transition dipoles [Deltaepsilon approximately Rda(-2) sin(2theta)]. Pulsed laser excitation of the stilbenediamide results in creation of the acceptor-donor radical ion pair, which decays via charge recombination. The dynamics of charge separation and charge recombination display an exponential distance dependence, similar to that observed previously for systems in which guanine serves as the electron donor. Unlike exciton coupling between the stilbenes, there is no apparent dependence of the charge-transfer rates upon the dihedral angle between donor and acceptor stilbenes. The introduction of a single G-C base pair between the donor and acceptor results in a change in the mechanism for charge separation from single step superexchange to hole hopping.


Scale-up of cell culture bioreactors using biomechatronic design.
Scale-up of cell culture bioreactors is a challenging engineering work that requires wide competence in cell biology, mechanical engineering and bioprocess design. In this article, a new approach for cell culture bioreactor scale-up is suggested that is based on biomechatronic design methodology. The approach differs from traditional biochemical engineering methodology by applying a sequential design procedure where the needs of the users and alternative design solutions are systematically analysed. The procedure is based on the biological and technical functions of the scaled-up bioreactor that are derived in functional maps, concept generation charts and scoring and interaction matrices. Basic reactor engineering properties, such as mass and heat transfer and kinetics are integrated in the procedure. The methodology results in the generation of alternative design solutions that are thoroughly ranked with help of the user needs. Examples from monoclonal antibodies and recombinant protein production illuminate the steps of the procedure. The methodology provides engineering teams with additional tools that can significantly facilitate the design of new production methods for cell culture processes.


An integrated microfluidic device for influenza and other genetic analyses.
An integrated microfluidic device capable of performing a variety of genetic assays has been developed as a step towards building systems for widespread dissemination. The device integrates fluidic and thermal components such as heaters, temperature sensors, and addressable valves to control two nanoliter reactors in series followed by an electrophoretic separation. This combination of components is suitable for a variety of genetic analyses. As an example, we have successfully identified sequence-specific hemagglutinin A subtype for the A/LA/1/87 strain of influenza virus. The device uses a compact design and mass production technologies, making it an attractive platform for a variety of widely disseminated applications.


Responses to exercise in normobaric hypoxia: comparison of elite and recreational ski mountaineers.
Hypoxia is known to reduce maximal oxygen uptake (VO(2max)) more in trained than in untrained subjects in several lowland sports. Ski mountaineering is practiced mainly at altitude, so elite ski mountaineers spend significantly longer training duration at altitude than their lower-level counterparts. Since acclimatization in hypobaric hypoxia is effective, the authors hypothesized that elite ski mountaineers would exhibit a VO2max decrement in hypoxia similar to that of recreational ski mountaineers. Eleven elite (E, Swiss national team) and 12 recreational (R) ski mountaineers completed an incremental treadmill test to exhaustion in normobaric hypoxia (H, 3000 m, F(1)O(2) 14.6% ± 0.1%) and in normoxia (N, 485 m, F(1)O(2) 20.9% ± 0.0%). Pulse oxygen saturation in blood (SpO(2)), VO(2max), minute ventilation, and heart rate were recorded. At rest, hypoxic ventilatory response was higher (P < .05) in E than in R (1.4 ± 1.9 vs 0.3 ± 0.6 L · min⁻¹ · kg⁻¹). At maximal intensity, SpO(2) was significantly lower (P < .01) in E than in R, both in N (91.1% ± 3.3% vs 94.3% ± 2.3%) and in H (76.4% ± 5.4% vs 82.3% ± 3.5%). In both groups, SpO(2) was lower (P < .01) in H. Between N and H, VO(2max) decreased to a greater extent (P < .05) in E than in R (-18% and -12%, P < .01). In E only, the VO(2max) decrement was significantly correlated with the SpO(2) decrement (r = .74, P < .01) but also with VO(2max) measured in N (r = .64, P < .05). Despite a probable better acclimatization to altitude, VO(2max) was more reduced in E than in R ski mountaineers, confirming previous results observed in lowlander E athletes.


Na+ pump in renal tubular cells is regulated by endogenous Na+-K+-ATPase inhibitor from hypothalamus.
Bovine hypothalamus contains a high affinity, specific, reversible inhibitor of mammalian Na+-K+-ATPase. Kinetic analysis using isolated membrane fractions showed binding and dissociation rates of the hypothalamic factor (HF) to be (like ouabain) relatively long (off rate = 60 min). To determine whether the kinetics of inhibition in intact cells might be more consistent with regulation of physiological processes in vivo, binding and dissociation reactions of HF in intact renal epithelial cells (LLC-PK1) were studied using 86Rb+ uptake and [3H]ouabain binding. As with membranes, a 60-min incubation with HF inhibited Na+-K+-ATPase in LLC-PK1 cells. In contrast to membrane studies, no prolonged incubation with LLC-PK1 was needed to observe inhibition of Na+-K+-ATPase. HF caused a 33% inhibition of ouabain-sensitive 86Rb+ influx within 10 min. Incubation of cells with HF followed by washout showed rapid reversal of pump inhibition and a doubling of pump activity. The dose-response curve for HF inhibition of LLC-PK1 86Rb+ uptake showed a sigmoidal shape consistent with an allosteric binding reaction. Thus HF is a potent regulator of Na+-K+-ATPase activity in intact renal cells, with binding and dissociation reactions consistent with relevant physiological processes.


Evaluation of teniposide (VM-26)-induced toxicity on mouse spermatogenesis by flow cytometry.
Alteration in the testicular weight and various germ cell populations was studied in male mice treated with different doses (0.05, 0.25, 0.5, 1.0 and 2.0 mg/kg b. wt.) of teniposide (VM-26) at various post-treatment time periods. Treatment of mice with different doses of teniposide did not significantly alter the testicular weights, irrespective of the drug dose used. Flow-cytometric analysis of germ cells of the untreated control mice testes revealed four distinct DNA peaks corresponding to elongated spermatids (HC), round spermatids (1C), spermatogonia and non-germ cells (2C) and primary spermatocytes (4C). The region between 2C and 4C peaks represents cells that are actively synthesizing DNA (S-phase cells). Treatment of mice with different doses of teniposide resulted in a significant depletion in the relative percentage of spermatogonia from day 2 to 35 post-treatment depending on the drug dose. DNA-synthesizing, i.e. S-phase, cells declined significantly at day 1 post-treatment and continued to decline up to day 70 post-treatment for all the drug doses studied, except 2 mg/kg drug dose at day 28 post-treatment. A significant decline in the relative percentage of primary spermatocytes (4C) was observed at day 7 that continued up to day 70 post-treatment depending on the drug dose. Round spermatids (1C) declined significantly at day 21 post-treatment after administration of 0.25--2.0 mg/kg VM-26. The relative percentage of elongated spermatids showed a significant decline at day 28 after 1 and 2 mg/kg drug treatment. These alterations in different germ-cell populations are reflected in the various germ-cell ratios. The 4C:2C ratio showed a significant decline at day 7 and 14 post-treatment after 1 and 2 mg/kg VM-26 treatment, while the 1C:2C ratio declined significantly at day 21 post-treatment in the mice treated with 0.5 and 2.0 mg/kg of VM-26. 4C:S-phase and 1C:4C ratios increased significantly from day 1 to 70 post-treatment, depending on the drug dose. Our study demonstrates that the treatment of mice with low doses of VM-26 exerts cytotoxic effects on various germ-cell populations.


Xander: employing a novel method for efficient gene-targeted metagenomic assembly.
Metagenomics can provide important insight into microbial communities. However, assembling metagenomic datasets has proven to be computationally challenging. Current methods often assemble only fragmented partial genes. We present a novel method for targeting assembly of specific protein-coding genes. This method combines a de Bruijn graph, as used in standard assembly approaches, and a protein profile hidden Markov model (HMM) for the gene of interest, as used in standard annotation approaches. These are used to create a novel combined weighted assembly graph. Xander performs both assembly and annotation concomitantly using information incorporated in this graph. We demonstrate the utility of this approach by assembling contigs for one phylogenetic marker gene and for two functional marker genes, first on Human Microbiome Project (HMP)-defined community Illumina data and then on 21 rhizosphere soil metagenomic datasets from three different crops totaling over 800 Gbp of unassembled data. We compared our method to a recently published bulk metagenome assembly method and a recently published gene-targeted assembler and found our method produced more, longer, and higher quality gene sequences. Xander combines gene assignment with the rapid assembly of full-length or near full-length functional genes from metagenomic data without requiring bulk assembly or post-processing to find genes of interest. HMMs used for assembly can be tailored to the targeted genes, allowing flexibility to improve annotation over generic annotation pipelines. This method is implemented as open source software and is available at https://github.com/rdpstaff/Xander_assembler.


Paeoniflorin inhibited the tumor invasion and metastasis in human hepatocellular carcinoma cells.
Evidence suggests that paeoniflorin may be involved in anticancer activities. Here, we have investigated the effects of paeoniflorin and correlative mechanisms on anti-invasion and anti-metastasis in human hepatocellular carcinoma (HCC) cell lines. In the current study, we have applied 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay to determine the proliferative effect of HepG2 and Bel-7402, two human hepatoma cell lines, and we have established a boyden chamber assay, a wound healing assay and cell adhesion assay to detect and quantify the invasion, metastasis and adhesion of both HepG2 and Bel-7402. In addition, we have analyzed the protein expression of matrix metalloproteinas (MMP)-9, E-cadherin (E-cad) and extracellular signal-regulated kinase (ERK) in both cell lines through western blot analysis. Paeoniflorin (6. 25-200 µM) had inhibitory effect on the growth of HepG2 and Bel-7402 cell lines, and reduced significantly invasion, metastasis and adhesion of HCC cell lines. In addition, paeoniflorin decreased the expression of MMP-9 and ERK in HepG2 and Bel-7402 cells, and increased expression of E-cad in both cell lines. Paeoniflorin is effective anti-metastatic and anti-invasive agent for suppressing HCC invasion and metastasis (Fig. 5, Ref. 30).


Anti-diabetic effect of purple corn extract on C57BL/KsJ db/db mice.
Recently, anthocyanins have been reported to have various biological activities. Furthermore, anthocyanin-rich purple corn extract (PCE) ameliorated insulin resistance and reduced diabetes-associated mesanginal fibrosis and inflammation, suggesting that it may have benefits for the prevention of diabetes and diabetes complications. In this study, we determined the anthocyanins and non-anthocyanin component of PCE by HPLC-ESI-MS and investigated its anti-diabetic activity and mechanisms using C57BL/KsJ db/db mice. The db/db mice were divided into four groups: diabetic control group (DC), 10 or 50 mg/kg PCE (PCE 10 or PCE 50), or 10 mg/kg pinitol (pinitol 10) and treated with drugs once per day for 8 weeks. During the experiment, body weight and blood glucose levels were measured every week. At the end of treatment, we measured several diabetic parameters. Compared to the DC group, Fasting blood glucose levels were 68% lower in PCE 50 group and 51% lower in the pinitol 10 group. Furthermore, the PCE 50 group showed 2- fold increased C-peptide and adiponectin levels and 20% decreased HbA1c levels, than in the DC group. In pancreatic islets morphology, the PCE- or pinitol-treated mice showed significant prevention of pancreatic β-cell damage and higher insulin content. Microarray analyses results indicating that gene and protein expressions associated with glycolysis and fatty acid metabolism in liver and fat tissues. In addition, purple corn extract increased the phosphorylation of AMP-activated protein kinase (AMPK) and decreased phosphoenolpyruvate carboxykinase (PEPCK), glucose 6-phosphatase (G6pase) genes in liver, and also increased glucose transporter 4 (GLUT4) expressions in skeletal muscle. Our results suggested that PCE exerted anti-diabetic effects through protection of pancreatic β-cells, increase of insulin secretion and AMPK activation in the liver of C57BL/KsJ db/db mice.


Radiotherapy in Combination With Cytokine Treatment.
Radiotherapy (RT) plays an important role in the management of cancer patients. RT is used in more than 50% of patients during the course of their disease in a curative or palliative setting. In the past decades it became apparent that the abscopal effect induced by RT might be dependent on the activation of immune system, and that the induction of immunogenic cancer cell death and production of danger-associated molecular patterns from dying cells play a major role in the radiotherapy-mediated anti-tumor efficacy. Therefore, the combination of RT and immunotherapy is of a particular interest that is reflected in designing clinical trials to treat patients with various malignancies. The use of cytokines as immunoadjuvants in combination with RT has been explored over the last decades as one of the immunotherapeutic combinations to enhance the clinical response to anti-cancer treatment. Here we review mainly the data on the efficacy of IFN-α, IL-2, IL-2-based immunocytokines, GM-CSF, and TNF-α used in combinations with various radiotherapeutic techniques in clinical trials. Moreover, we discuss the potential of IL-15 and its analogs and IL-12 cytokines in combination with RT based on the efficacy in preclinical mouse tumor models.


Novel method to characterize CYP21A2 in Florida patients with congenital adrenal hyperplasia and commercially available cell lines.
Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder and affects approximately 1 in 15,000 births in the United States. CAH is one of the disorders included on the Newborn Screening (NBS) Recommended Uniform Screening Panel. The commonly used immunological NBS test is associated with a high false positive rate and there is interest in developing second-tier assays to increase screening specificity. Approximately 90% of the classic forms of CAH, salt-wasting and simple virilizing, are due to mutations in the CYP21A2 gene. These include single nucleotide changes, insertions, deletions, as well as chimeric genes involving CYP21A2 and its highly homologous pseudogene CYP21A1P. A novel loci-specific PCR approach was developed to individually amplify the CYP21A2 gene, the nearby CYP21A1P pseudogene, as well as any 30 kb deletion and gene conversion mutations, if present, as single separate amplicons. Using commercially available CAH positive specimens and 14 families with an affected CAH proband, the single long-range amplicon approach demonstrated higher specificity as compared to previously published methods.


Cancer survival in patients from a hospital-based cancer registry, China.
Purpose: There are few reports on survival rate analysis from hospital-based cancer registries (HBCR) in China, although the National Center of Cancer Registry of China has launched such an effort with the mission to expand the scope of registration and follow-up. Our study aimed to evaluate survival and outcomes of cancer patients from a HBCR in eastern China. Methods: Active and passive follow-up methods were used to obtain information on survival status for all patients from Qidong City and Haimen City in the databases of our hospital-based registrations from 2002 to 2014. Censor time for survival was 31st March, 2016. Survival probability was estimated using the life-table method with SPSS Statistics software, and comparison of significant differences in survival rates was tested by Wilcoxon (Gehan) statistic. Results: The outcomes of 5010 patients were identified in the follow-up for 5244 cases from Qidong and Haimen, with a follow-up rate of 95.65%, and a rate of lost to follow-up of 4.35%. The 1-, 3-, 5-, and 10-year observed survival (OS) rate in all-combined cancer sites were 59.80%, 37.70%, 30.82%, and 22.60%, respectively. The top 10 cancer sites in rank were cancers of lung, esophagus, liver, cervix, stomach, breast, colon-rectum, non-Hodgkin's lymphoma, nasopharynx, and ovary, with 5-year OS rates of 12.63%, 19.62%, 11.69%, 66.61%, 21.35%, 59.43%, 36.36%, 37.03%, 48.95% and 36.17%, respectively. Females experienced better survival than males for lung, esophageal, liver, nasopharyngeal and pancreatic cancers (P<0.05), but not for other sites (P>0.05). A significant difference was also found between males and females when all-sites were combined (P<0.01). There are significant differences (P<0.05) between the 2015 patients (from Qidong) and the 3001 patients (from Haimen) with 5-year OS rates of 32.72% vs 29.57%; no significant differences were found for 5-year OS rates for individual cancer sites (P>0.05) except for liver (P=0.0005) and ovary (P=0.0460) between the two cities. Younger patients had better prognosis, but significance was only seen in cervical (P=0.0102) and nasopharyngeal (P=0.0305) cancers. Conclusion: The survival rates of each site or of all sites-combined in this setting are consistent with those elsewhere in China and abroad. Discrepancies in overall survival could be affected by the proportion of sites with or without better prognosis. Hospital-based cancer survival is a better index to evaluate outcomes that reflect the levels of comprehensive treatment and improvement of medical and health services.


Clinical Appraisal of Fosfomycin in the Era of Antimicrobial Resistance.
Fosfomycin is recommended as one of the first-line agents for treatment of urinary tract infections (UTIs) in the latest guidelines endorsed by the Infectious Diseases Society of America (IDSA) and the European Society for Clinical Microbiology and Infectious Diseases (ESCMID). We evaluated the use of fosfomycin among inpatients at a tertiary care hospital between 2009 and 2013. UTI cases were defined using physician diagnosis and the National Healthcare Safety Network (NHSN) surveillance definitions. The number of patients treated with fosfomycin increased from none in 2009 to 391 in 2013. Among 537 patients who received fosfomycin for any indication during this period, UTI was the most common indication (74%), followed by asymptomatic bacteriuria (10%). All except 19 patients received a single dose of fosfomycin. Escherichia coli was the most common organism involved (52%). For 119 patients with UTIs, after exclusion of those with negative urine culture results, negative urinalysis results, receipt of additional agents, or indeterminate clinical outcomes, the clinical success rate at 48 h was 74.8%. Of 89 patients who met the criteria for NHSN-defined UTIs, 89.9% had successful outcomes. Recurrent infections occurred in 4.3% of cases, and mild adverse events were observed in 2.0%. All 100 randomly selected extended-spectrum β-lactamase (ESBL)-producing E. coli clinical isolates from this period were susceptible to fosfomycin. In conclusion, the use of fosfomycin has increased substantially since implementation of the updated guidelines at this hospital. Fosfomycin was used mainly for the treatment of physician-diagnosed UTIs, and the clinical outcomes were generally favorable. Fosfomycin maintained activity against E. coli despite the increased use of the agent.


Feasibility Validation on Healthy Adults of a Novel Active Vibrational Sensing Based Ankle Band for Ankle Flexion Angle Estimation.
Goal: In this paper, we introduced a novel ankle band with a vibrational sensor that can achieve low-cost ankle flexion angle estimation, which can be potentially used for automated ankle flexion angle estimation in home-based foot drop rehabilitation scenarios. Methods: Previous ankle flexion angle estimation methods require either professional knowledge or specific equipment and lab environment, which is not feasible for foot drop patients to achieve accurate measurement by themselves in a home-based scenario. To solve the above problems, a prototype was developed based on the assumption that the echo of a vibration signal on the tibialis anterior had different acoustic impedance distribution. By analyzing the frequency spectrum of the echo, the ankle flexion angle can be estimated. Therefore, a surface transducer was utilized to generate frequency-varying active vibration, and a contact microphone was utilized to capture the echo. A portable analog signal processing hub drove the transducer, and was used for echo signal collection from the microphone. Finally, a Random Forest regression model was applied to estimate the ankle flexion angle based on the spectrum amplitude of the echo. Results: Five healthy subjects were recruited in the experiment. The regression estimation error is 4.16 degrees, and the R2 is 0.81. Conclusions: These results demonstrate the feasibility of the proposed ankle band for accurate ankle flexion angle estimation.


Implementation and spread of interventions into the multilevel context of routine practice and policy: implications for the cancer care continuum.
The promise of widespread implementation of efficacious interventions across the cancer continuum into routine practice and policy has yet to be realized. Multilevel influences, such as communities and families surrounding patients or health-care policies and organizations surrounding provider teams, may determine whether effective interventions are successfully implemented. Greater recognition of the importance of these influences in advancing (or hindering) the impact of single-level interventions has motivated the design and testing of multilevel interventions designed to address them. However, implementing research evidence from single- or multilevel interventions into sustainable routine practice and policy presents substantive challenges. Furthermore, relatively few multilevel interventions have been conducted along the cancer care continuum, and fewer still have been implemented, disseminated, or sustained in practice. The purpose of this chapter is, therefore, to illustrate and examine the concepts underlying the implementation and spread of multilevel interventions into routine practice and policy. We accomplish this goal by using a series of cancer and noncancer examples that have been successfully implemented and, in some cases, spread widely. Key concepts across these examples include the importance of phased implementation, recognizing the need for pilot testing, explicit engagement of key stakeholders within and between each intervention level; visible and consistent leadership and organizational support, including financial and human resources; better understanding of the policy context, fiscal climate, and incentives underlying implementation; explication of handoffs from researchers to accountable individuals within and across levels; ample integration of multilevel theories guiding implementation and evaluation; and strategies for long-term monitoring and sustainability.


Direct effect of insulin on albumin gene expression in primary cultures of rat hepatocytes.
The purpose of this study was to identify a cell culture system in which the role of insulin in regulating albumin gene expression could be investigated. The system selected was rat hepatocytes maintained in primary culture in a chemically defined, serum-free medium. Under control conditions albumin secretion was nearly the same as the rate recorded in vivo and in perfused liver and was reasonably well maintained during 8 days of culture. Deletion of insulin from the culture medium for 3-6 days resulted in 40-60% reductions in albumin secretion. Furthermore, albumin secretion relative to the rate of total protein synthesis was reduced by approximately 50% as a result of insulin deficiency. Readdition of the hormone to insulin-deficient cultures restored secretion to the control rate. A maximal effect of insulin was observed within 3 days after readdition of the hormone, and a half-maximal response was obtained with a hormone concentration of approximately 3.0 nM. The relative abundance of albumin mRNA, as measured by solution hybridization using a complementary DNA probe, responded in a parallel fashion to the changes in albumin secretion. Thus rat hepatocytes maintained under appropriate culture conditions reflect the effects of diabetes and insulin treatment on albumin gene expression observed in vivo and provide an excellent model system in which to study the mechanism(s) of insulin action.


A two-compartment model of VEGF distribution in the mouse.
Vascular endothelial growth factor (VEGF) is a key regulator of angiogenesis--the growth of new microvessels from existing microvasculature. Angiogenesis is a complex process involving numerous molecular species, and to better understand it, a systems biology approach is necessary. In vivo preclinical experiments in the area of angiogenesis are typically performed in mouse models; this includes drug development targeting VEGF. Thus, to quantitatively interpret such experimental results, a computational model of VEGF distribution in the mouse can be beneficial. In this paper, we present an in silico model of VEGF distribution in mice, determine model parameters from existing experimental data, conduct sensitivity analysis, and test the validity of the model. The multiscale model is comprised of two compartments: blood and tissue. The model accounts for interactions between two major VEGF isoforms (VEGF(120) and VEGF(164)) and their endothelial cell receptors VEGFR-1, VEGFR-2, and co-receptor neuropilin-1. Neuropilin-1 is also expressed on the surface of parenchymal cells. The model includes transcapillary macromolecular permeability, lymphatic transport, and macromolecular plasma clearance. Simulations predict that the concentration of unbound VEGF in the tissue is approximately 50-fold greater than in the blood. These concentrations are highly dependent on the VEGF secretion rate. Parameter estimation was performed to fit the simulation results to available experimental data, and permitted the estimation of VEGF secretion rate in healthy tissue, which is difficult to measure experimentally. The model can provide quantitative interpretation of preclinical animal data and may be used in conjunction with experimental studies in the development of pro- and anti-angiogenic agents. The model approximates the normal tissue as skeletal muscle and includes endothelial cells to represent the vasculature. As the VEGF system becomes better characterized in other tissues and cell types, the model can be expanded to include additional compartments and vascular elements.


Interaction between electrical stimulation, protein coating and matrix elasticity: a complex effect on muscle fibre maturation.
In skeletal muscle tissue engineering, it remains a challenge to produce mature, functional muscle tissue. Mimicking the in vivo niche in in vitro culture might overcome this problem. Niche components include, for example, extracellular matrix proteins, neighbouring cells, growth factors and physical factors such as the elasticity of the matrix. Previously, we showed the effects of matrix stiffness and protein coating on proliferation and differentiation of muscle progenitor cells in a two-dimensional (2D) situation. In the present study we have investigated the additional effect of electrical stimulation. More precisely, we investigated the effect of electrical stimulation on primary myoblast maturation when cultured on top of Matrigel™ - or laminin-coated substrates with varying elasticities. The effect of electrical stimulation on differentiation and maturation was found to be dependent on coating and stiffness. Although electrical stimulation enhanced myoblast maturation, the effect was mild. We therefore conclude that, with the current regimen, electrical stimulation is not essential to create functional, mature muscle tissue.


Responses in macaque visual area V4 following inactivation of the parvocellular and magnocellular LGN pathways.
A substantial body of evidence has suggested that signals transmitted through the magnocellular and parvocellular subdivisions of the LGN remain largely segregated in visual cortex. This hypothesis can be tested directly by selectively blocking transmission through either the magnocellular or parvocellular layers with small injections of lidocaine or GABA while recording cortical responses to a visual stimulus. In a previous study, we found that responses in the middle temporal visual area (MT) were almost always greatly reduced by blocks of magnocellular LGN, but that few MT neurons were affected by parvocellular blocks. In the present study, we have examined magnocellular and parvocellular contributions to area V4, which lies at the same level of processing in the cortical hierarchy as does MT and is thought to be a major recipient of parvocellular input. We found that inactivation of parvocellular layers usually resulted in a moderate reduction of visual responses (median reduction, 36%). However, comparable reductions in V4 responses were also seen following magnocellular blocks (median reduction, 47%). Directionally selective responses in V4 were not found to depend specifically on either subdivision. We conclude that area V4, unlike MT, receives strong input from both subdivisions of the LGN. These results suggest that the relationship between the subcortical magnocellular and parvocellular pathways and the parietal and temporal streams of processing in cortex is not one-to-one.


Performance of Different Diagnostic Criteria for Familial Mediterranean Fever in Children with Periodic Fevers: Results from a Multicenter International Registry.
Our aims were to validate the pediatric diagnostic criteria in a large international registry and to compare them with the performance of previous criteria for the diagnosis of familial Mediterranean fever (FMF). Pediatric patients with FMF from the Eurofever registry were used for the validation of the existing criteria. The other periodic fevers served as controls: mevalonate kinase deficiency (MKD), tumor necrosis factor receptor-associated periodic syndrome (TRAPS), cryopyrin-associated periodic syndrome (CAPS), aphthous stomatitis, pharyngitis, adenitis syndrome (PFAPA), and undefined periodic fever from the same registry. The performances of Tel Hashomer, Livneh, and the Yalcinkaya-Ozen criteria were assessed. The FMF group included 339 patients. The control group consisted of 377 patients (53 TRAPS, 45 MKD, 32 CAPS, 160 PFAPA, 87 undefined periodic fevers). Patients with FMF were correctly diagnosed using the Yalcinkaya-Ozen criteria with a sensitivity rate of 87.4% and a specificity rate of 40.7%. On the other hand, Tel Hashomer and Livneh criteria displayed a sensitivity of 45.0 and 77.3%, respectively. Both of the latter criteria displayed a better specificity than the Yalcinkaya-Ozen criteria: 97.2 and 41.1% for the Tel Hashomer and Livneh criteria, respectively. The overall accuracy for the Yalcinkaya-Ozen criteria was 65 and 69.6% (using 2 and 3 criteria), respectively. Ethnicity and residence had no effect on the performance of the Yalcinkaya-Ozen criteria. The Yalcinkaya-Ozen criteria yielded a better sensitivity than the other criteria in this international cohort of patients and thus can be used as a tool for FMF diagnosis in pediatric patients from either the European or eastern Mediterranean region. However, the specificity was lower than the previously suggested adult criteria.


Sequence variations in the public human genome data reflect a bottlenecked population history.
Single-nucleotide polymorphisms (SNPs) constitute the great majority of variations in the human genome, and as heritable variable landmarks they are useful markers for disease mapping and resolving population structure. Redundant coverage in overlaps of large-insert genomic clones, sequenced as part of the Human Genome Project, comprises a quarter of the genome, and it is representative in terms of base compositional and functional sequence features. We mined these regions to produce 500,000 high-confidence SNP candidates as a uniform resource for describing nucleotide diversity and its regional variation within the genome. Distributions of marker density observed at different overlap length scales under a model of recombination and population size change show that the history of the population represented by the public genome sequence is one of collapse followed by a recent phase of mild size recovery. The inferred times of collapse and recovery are Upper Paleolithic, in agreement with archaeological evidence of the initial modern human colonization of Europe.


Hyperinduction of host beta interferon by a Listeria monocytogenes strain naturally overexpressing the multidrug efflux pump MdrT.
Many pathogens regulate or modify their immune-stimulating ligands to avoid detection by their infected hosts. Listeria monocytogenes, a facultative intracellular bacterial pathogen, interacts with multiple components of mammalian innate immunity during its infection cycle. During replication within the cytosol of infected cells, L. monocytogenes utilizes two multidrug efflux pumps, MdrM and MdrT, to secrete the small nucleic acid second messenger cyclic-di-AMP (c-di-AMP). Host recognition of c-di-AMP triggers the production of type I interferons, including beta interferon (IFN-β), which, surprisingly, promote L. monocytogenes virulence. In this study, we have examined the capacity of multiple laboratory and clinical isolates of L. monocytogenes to stimulate host production of IFN-β. We have identified the L. monocytogenes strain LO28 as able to hyperinduce IFN-β production in infected cells ∼30-fold more than the common laboratory clone L. monocytogenes strain 10403S. Genomic analyses determined that LO28 contains a naturally occurring loss-of-function allele of the transcriptional regulator BrtA and correspondingly derepresses expression of MdrT. Surprisingly, while derepression of MdrT resulted in hyperstimulation of IFN-β, it results in significant attenuation in multiple mouse models of infection. While type I interferons may promote L. monocytogenes virulence, this study demonstrates that unregulated expression of the c-di-AMP-secreting efflux pump MdrT significantly restricts virulence in vivo by an unknown mechanism.


Is increased residual shank length a competitive advantage for elite transtibial amputee long jumpers?
The purpose of this study was to evaluate the extent to which residual shank length affects long jump performance of elite athletes with a unilateral transtibial amputation. Sixteen elite, male, long jumpers with a transtibial amputation were videoed while competing in major championships (World Championships 1998, 2002 and Paralympic Games, 2004). The approach, take-off, and landing of each athlete's best jump was digitized to determine residual and intact shank lengths, jump distance, and horizontal and vertical velocity of center of mass at touchdown. Residual shank length ranged from 15 cm to 38 cm. There were weak, nonsignificant relationships between residual shank length and (a) distance jumped (r = 0.30), (b) horizontal velocity (r = 0.31), and vertical velocity (r = 0.05). Based on these results, residual shank length is not an important determinant of long jump performance, and it is therefore appropriate that all long jumpers with transtibial amputation compete in the same class. The relationship between residual shank length and key performance variables was stronger among athletes that jumped off their prosthetic leg (N = 5), and although this result must be interpreted cautiously, it indicates the need for further research.


Determinants of male involvement in maternal and child health services in sub-Saharan Africa: a review.
Male participation is a crucial component in the optimization of Maternal and Child Health (MCH) services. This is especially so where prevention strategies to decrease Mother-to-Child Transmission (MTCT) of Human Immunodeficiency Virus (HIV) are sought. This study aims to identify determinants of male partners' involvement in MCH activities, focusing specifically on HIV prevention of maternal to child transmission (PMTCT) in sub-Saharan Africa. Literature review was conducted using the following data bases: Pubmed/MEDLINE; CINAHL; EMBASE; COCHRANE; Psych INFORMATION and the websites of the International AIDS Society (IAS), the International AIDS Conference and the International Conference on AIDS in Africa (ICASA) 2011. We included 34 studies in this review, which reported on male participation in MCH and PMTCT services. The majority of studies defined male participation as male involvement solely during antenatal HIV testing. Other studies defined male involvement as any male participation in HIV couple counseling. We identified three main determinants for male participation in PMTCT services: 1) Socio-demographic factors such as level of education, income status; 2) health services related factors such as opening hours of services, behavior of health providers and the lack of space to accommodate male partners; and 3) Sociologic factors such as beliefs, attitudes and communication between men and women. There are many challenges to increase male involvement/participation in PMTCT services. So far, few interventions addressing these challenges have been evaluated and reported. It is clear however that improvement of antenatal care services by making them more male friendly, and health education campaigns to change beliefs and attitudes of men are absolutely needed.


Comparison of cord blood and peripheral blood mononuclear cells as targets for viral isolation and drug sensitivity studies involving human immunodeficiency virus type 1.
We have shown that umbilical cord blood mononuclear cells (CBMC) are at least as sensitive as peripheral blood mononuclear cells (PBMC) for isolation of human immunodeficiency virus type 1 from the PBMC of infected individuals. Viral replication was more efficiently monitored by a p24 antigen capture assay than by a viral reverse transcriptase test, regardless of whether CBMC or PBMC were employed. We also found that CBMC and PBMC yielded similar results with regard to the susceptibility profiles of both wild-type and drug-resistant variants of human immunodeficiency virus type 1 for 3'-azido-3'-deoxythymidine, 2',3'-dideoxycytidine, and the (-) enantiomer of 2',3'-dideoxy-3'-thiacytidine. Finally, viruses isolated on CBMC could be routinely grown on PBMC and vice versa.


Chemical quality and regulatory compliance of drinking water in Iceland.
Assuring sufficient quality of drinking water is of great importance for public wellbeing and prosperity. Nations have developed regulatory system with the aim of providing drinking water of sufficient quality and to minimize the risk of contamination of the water supply in the first place. In this study the chemical quality of Icelandic drinking water was evaluated by systematically analyzing results from audit monitoring where 53 parameters were assessed for 345 samples from 79 aquifers, serving 74 water supply systems. Compliance to the Icelandic Drinking Water Regulation (IDWR) was evaluated with regard to parametric values, minimum requirement of sampling, and limit of detection. Water quality compliance was divided according to health-related chemicals and indicators, and analyzed according to size. Samples from few individual locations were benchmarked against natural background levels (NBLs) in order to identify potential pollution sources. The results show that drinking compliance was 99.97% in health-related chemicals and 99.44% in indicator parameters indicating that Icelandic groundwater abstracted for drinking water supply is generally of high quality with no expected health risks. In 10 water supply systems, of the 74 tested, there was an indication of anthropogenic chemical pollution, either at the source or in the network, and in another 6 water supplies there was a need to improve the water intake to prevent surface water intrusion. Benchmarking against the NBLs proved to be useful in tracing potential pollution sources, providing a useful tool for identifying pollution at an early stage.


Intracranial temperature elevation from diagnostic ultrasound.
Tissues of the central nervous system are sensitive to damage by physical agents, such as heat and ultrasound. Exposure to pulsed spectral Doppler ultrasound can significantly heat biologic tissue because of the relatively high intensities used and the need to hold the beam stationary during examinations. This has significant implications for sensitive neural tissue such as that exposed during spectral Doppler flow studies of fetal cerebral vessels. Recent changes in the FDA regulation allow delivery of almost eight times higher intensity into the fetal brain by ultrasound devices that incorporate an approved real-time output display in their design. In this situation, ultrasound users are expected to assess the risk/benefit ratio based on their interpretation of equipment output displays (including the thermal index, TI) and an understanding of the significance of biologic effects. To assist in the assessment of potential thermally mediated bioeffects, a number of conclusions can be drawn from the published scientific literature: the amount of ultrasound-induced intracranial heating increases with gestational age and the development of fetal bone; pulsed spectral Doppler ultrasound can produce biologically significant heating in the fetal brain; the rate of heating near bone is rapid, with approximately 75% of the maximum heating occurring within 30 s; blood flow has minimal cooling effect on ultrasound-induced heating of the brain when insonated with narrow focused clinical beams; the threshold for irreversible damage in the developing embryo and fetal brain is exceeded when a temperature increase of 4 degrees C is maintained for 5 min; an ultrasound exposure that produces a temperature increase of up to 1.5 degrees C in 120 s does not elicit measurable electrophysiologic responses in fetal brain; for some exposure conditions, the thermal index (TI), as used in the FDA-approved output display standard, underestimates the extent of ultrasound-induced intracranial temperature increase.


Molecular analysis of the NSP4 and VP6 genes of rotavirus strains recovered from hospitalized children in Rio de Janeiro, Brazil.
Group A rotaviruses are the main cause of acute gastroenteritis in children throughout the world. The two outer capsid proteins, VP4 and VP7, define the P and G genotypes, respectively. Rotaviruses with P[8]G1, P[4]G2, P[8]G3 and P[8]G4 genotypes are predominant in infecting humans and the G9 genotype is emerging in most continents as the fifth most common G type worldwide. The inner capsid protein VP6 is responsible for subgroup (SG) specificities, allowing classification of rotaviruses into SG I, SG II, SG I+II and SG non-I-non-II. The non-structural protein 4 (NSP4) encoded by segment 10 has a role in viral morphogenesis and five genetic groups have been described, NSP4 genotypes A-E. The aim of this investigation was to characterize the NSP4 and VP6 genes of rotavirus strains recovered from hospitalized children. Thirty rotavirus strains were submitted to RT-PCR followed by sequencing and phylogenetic analysis. Among the different G and P genotype combinations, two distinct genetic groups could be recognized for the NSP4 gene. Twenty-eight clustered with NSP4 genotype B. The two P[4]G2 strains fell into NSP4 genotype A and clustered distinctly, with a 100 % bootstrap value. The strains distinguished within a group were closely related to each other at the nucleotide and amino acid levels. A phylogenetic tree was constructed for the VP6 gene including the human strains RMC100, E210, Wa, US1205 and 1076, and the animal strains Gott, NCDV, SA-11, FI-14 and EW. This is the first report on Brazilian rotavirus strains describing NSP4 genotype A strains associated with VP6 SG I, and NSP4 genotype B strains associated with VP6 SG II.


[ANTHROPOMETRIC CHILEAN TABLE TENNIS PLAYERS OF COMPETITIVE FEATURES].
The aim of the study was to characterize the anthropometric profile and somatotype of a sample of 50 players table tennis competitive with an average age 21.6 (± 3.1) years belonging to the Chilean team and institutions of higher education in the region of Valparaiso. The evaluation was conducted under the protocol marking the International Society for the Advancement of Kinanthropometry (ISAK) for the measurement procedure 25 restricted profile variables described by Drinkwater, Norton and Olds. Order to determine the body composition, fat, muscle, bone, skin and tissue residual was considered, using the equations proposed by Kerr. The body shape is characterized through somatotype method proposed by Carter. The sample was divided into 4 groups; Chilean Selection, Traditional Private Universities, State Universities and Private Universities Traditional Nontraditional. Regarding body composition; the Chilean team has the highest values of muscle tissue (45.6 ± 1.7%) and the lowest values of adipose tissue (25.2 ± 1.8%), also presenting lesser value in the Σ 6 skinfolds (mm) . The results showed no significant differences between groups in the aforementioned variables. In general somatotype compared by analyzing SANOVA no significant differences between groups (p = 0.409) was observed. The results show a biotype with such a characterization of endo-mesomorph with average values (4,1-4,9-1,8). This study provides updated data biotypological reference for this sport that can be used for decision-making.


In vitro metabolism and metabolic effects of ajulemic acid, a synthetic cannabinoid agonist.
Ajulemic acid is a synthetic analog of Δ(8)-THC-11-oic acid, the terminal metabolite of Δ(8)-THC. Unlike Δ(9)-THC, the psychoactive principle of Cannabis, it shows potent anti-inflammatory action and has minimal CNS cannabimimetic activity. Its in vitro metabolism by hepatocytes from rats, dogs, cynomolgus monkeys and humans was studied and the results are reported here. Five metabolites, M1 to M5, were observed in human hepatocyte incubations. One metabolite, M5, a glucuronide, was observed in the chromatogram of canine hepatocyte incubations. In monkey hepatocyte incubations, M5 was observed in the chromatograms of both the 120 and 240 min samples, trace metabolite M1 (side-chain hydroxyl) was observed in the 120 min samples, and trace metabolite M4 (side-chain dehydrogenation) was observed in the 240 min samples. No metabolites were found in the rat hepatocyte incubations. Unchanged amounts of ajulemic acid detected after the 2-h incubation were 103%, 90%, 86%, and 83% for rat, dog, monkey, and human hepatocytes, respectively. Additional studies were done to ascertain if ajulemic acid can inhibit the activities of five principal human cytochrome P450 isozymes; CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4/5. In contrast to the phytocannabinoids Δ(9)-THC and CBD, no significant inhibition of cytochrome activity was observed. These data further support the conclusions reached in earlier reports on ajulemic acid's high margin of safety and suggest that it undergoes minimal metabolism and is not likely to interfere with the normal metabolism of drugs or endogenous substances.


Percutaneous US-guided renal biopsy: a retrospective study comparing the 16-gauge end-cut and 14-gauge side-notch needles.
Assess glomerular yield and safety profile of two different types of needles for percutaneous ultrasound-guided kidney biopsy. Over 24 months, 121 ultrasonographic ultrasound-guided renal biopsies were performed on native kidneys of 121 adults: 66 with 16-gauge, 29-mm end-cut (BioPince) needles and 55 with 14-gauge, 1.9-mm side-notch (Tru-Cut) needles. The mean number of complete glomeruli harvested per biopsy was 21.0 and 19.3, respectively, and the mean number of core samples required to obtain a satisfactory biopsy was 1.8 and 2.6, respectively. The ratio of glomeruli harvested to core samples needed with the end-cut needle was 58% greater than that with the side-notch needles (11.7 vs 7.4, respectively; difference of 4.3; 95% confidence interval: 2.0, 6.8). Procedures performed with end-cut needles were associated with fewer major complications (1.5% vs 7.3% with side-notch needles). Compared to the 14-g Tru-cut needle, the 16-g end-cut needle provided better glomerular yield per core sample, required fewer cores for satisfactory tissue specimen, and resulted in fewer major complications.


Prevalence of asthma and COPD in general practice in 1992: has it changed since 1977?
Asthma and COPD are common diseases of the airways which are mainly diagnosed and treated in general practice. Various studies have reported an increase in the morbidity of asthma and COPD. There are two possible reasons for such an increase. The first is an apparent increase caused by using different criteria in defining asthma/COPD, and by increased awareness of the disease by doctors and patients. The second is a real increase caused by an increase in the prevalence of airway pathology. The aim of this study was to determine the cause of the observed increase in morbidity. In 1977, a sample of 2328 adults from the general population were screened for asthma and COPD. Those screened were then divided into five sub-groups (grades 1-5), according to severity of: (1) respiratory symptoms; and (2) loss in FEV1. The number of patients who were not known to the general practitioner prior to the screening as having asthma or COPD grades 1-5 were also assessed. In 1992, we studied a different sample of 1184 adults of the general population in the same area. We used the same criteria as in 1977 to analyse our results. The number of patients not known to the general practitioner prior to the screening was also studied. The overall prevalence (grades 1-5) of asthma and COPD has increased from +/- 19% in 1977 to +/- 31% in 1992 (range 21-42). The main reason for this is an increase in prevalence of very mild to moderate asthma and COPD (grades 1-3) from 17% in 1977 to 27% in 1992. The prevalence of severe cases (grades 4-5) increased from 2% in 1977 to 4% in 1992. In 1992, around 65% of the patients were not known to the general practitioner as having any grade of asthma or COPD. This was only slightly lower than the 72% in 1977. All patients with a severe disease (grade 5) were known to the general practitioner. There is a real increase in the prevalence of asthma and COPD, caused predominantly by an increase in the number of mild cases. The percentage of patients not known to the GP were predominantly mild cases.


The project MinE databrowser: bringing large-scale whole-genome sequencing in ALS to researchers and the public.
Amyotrophic lateral sclerosis (ALS) is a rapidly progressive fatal neurodegenerative disease affecting one in 350 people. The aim of Project MinE is to elucidate the pathophysiology of ALS through whole-genome sequencing at least 15,000 ALS patients and 7500 controls at 30× coverage. Here, we present the Project MinE data browser ( databrowser.projectmine.com ), a unique and intuitive one-stop, open-access server that provides detailed information on genetic variation analyzed in a new and still growing set of 4366 ALS cases and 1832 matched controls. Through its visual components and interactive design, the browser specifically aims to be a resource to those without a biostatistics background and allow clinicians and preclinical researchers to integrate Project MinE data into their own research. The browser allows users to query a transcript and immediately access a unique combination of detailed (meta)data, annotations and association statistics that would otherwise require analytic expertise and visits to scattered resources.


High stability configuration for an acoustooptic heterodyne rf spectrum analyzer using slow shear TeO(2).
A triangular, common-path interferometric rf spectrum analyzer arrangement has been demonstrated which has the property of high immunity to ambient vibration. Calculations show the TCPI arrangement, with two counter-propagating laser beams, is highly angle compensated and alignment stable when components are disturbed in position or orientation. Proper light polarization and Bragg cell geometry allows one Bragg cell to diffract primarily the clockwise beam and the other Bragg cell the counterclockwise beam. A birefringent quartz wedge shifts the heterodyne signal to a nonzero IF carrier frequency. Measured ambient phase noise is less than two degrees and calculated values indicate that further improvement may be possible.


A systematic review and thematic synthesis exploring how a previous experience of physical activity influences engagement with cardiac rehabilitation.
Exercise-based cardiac rehabilitation is recognised internationally as an effective therapy to improve quality of life and reduce the risk of hospital readmission for individuals diagnosed with acute coronary syndrome. Despite this, half of eligible individuals choose not to engage and the main reason is lack of interest. Furthermore, prior to attending, 40% of eligible individuals report meeting physical activity guidelines. It is unclear whether this influences decisions about engagement. The aim of this review is to examine systematically qualitative evidence describing patients' perceptions and experiences, and synthesise what is known about how a previous experience of physical activity in adults diagnosed with acute coronary syndrome influences engagement with physical activity during cardiac rehabilitation. A systematic review and thematic synthesis was conducted of primary qualitative studies to examine peer-reviewed literature published between1990 and 2017, accessed from database searches of MEDLINE, CINHAL, PsycINFO and Embase. The initial search produced 486 studies, and of these 12 relevant studies were included in this review. Studies included 388 participants from six countries. For previously active individuals, communication factors, self-perceptions of an exercise identity and experience of cardiac rehabilitation influence engagement in physical activity during cardiac rehabilitation. In adults diagnosed with acute coronary syndrome, communication post event and during cardiac rehabilitation is a source of self-appraisal and creates expectations of cardiac rehabilitation. In addition, perceptions of an exercise identity and experience of exercise-based cardiac rehabilitation influence decisions about engagement. To improve uptake and adherence, health professionals should consider previous physical activity levels and tailor information to optimise physical activity post event.


Exploring the effect of microdosing psychedelics on creativity in an open-label natural setting.
Taking microdoses (a mere fraction of normal doses) of psychedelic substances, such as truffles, recently gained popularity, as it allegedly has multiple beneficial effects including creativity and problem-solving performance, potentially through targeting serotonergic 5-HT2A receptors and promoting cognitive flexibility, crucial to creative thinking. Nevertheless, enhancing effects of microdosing remain anecdotal, and in the absence of quantitative research on microdosing psychedelics, it is impossible to draw definitive conclusions on that matter. Here, our main aim was to quantitatively explore the cognitive-enhancing potential of microdosing psychedelics in healthy adults. During a microdosing event organized by the Dutch Psychedelic Society, we examined the effects of psychedelic truffles (which were later analyzed to quantify active psychedelic alkaloids) on two creativity-related problem-solving tasks: the Picture Concept Task assessing convergent thinking and the Alternative Uses Task assessing divergent thinking. A short version of the Ravens Progressive Matrices task assessed potential changes in fluid intelligence. We tested once before taking a microdose and once while the effects were expected to be manifested. We found that both convergent and divergent thinking performance was improved after a non-blinded microdose, whereas fluid intelligence was unaffected. While this study provides quantitative support for the cognitive-enhancing properties of microdosing psychedelics, future research has to confirm these preliminary findings in more rigorous placebo-controlled study designs. Based on these preliminary results, we speculate that psychedelics might affect cognitive metacontrol policies by optimizing the balance between cognitive persistence and flexibility. We hope this study will motivate future microdosing studies with more controlled designs to test this hypothesis.


Intra-storm study of solute chemical composition of overland flow water in two agricultural fields.
Few studies have assessed the transport of dissolved nutrients at the field scale under natural rainfall conditions. Hysteresis between dissolved nutrients and discharge behavior can complicate such assessments and this effect has only been examined qualitatively. In this study, we investigated factors contributing to short-term variations of dissolved cation (Ca, Mg, Na, and K) and anion (soluble reactive phosphorus [SRP], NO3, and SO4) concentrations in runoff water and developed a quantitative method to study their hysteretic behavior. Within-storm variations of dissolved nutrient concentrations were determined in two agricultural fields during four natural rainfall events along with discharge, sediment, antecedent soil water conditions, and nutrient contents. For each event, nutrient loads were plotted against discharge during the rising and falling limb of the runoff hydrograph. The resulting hysteresis curves were characterized by an index H, which is the ratio between the integrated areas under the rising and falling curves of the hydrograph. Results showed that nutrient concentrations increased with time during each event. Counterclockwise (H < 1) hysteresis, occurring when the falling limb had larger loads, was found when soils were initially dry whereas clockwise hysteresis (H > 1) was associated with prior wet soil conditions. Two hypotheses are suggested to explain these variations. First, suspended sediments could have acted as a sink for dissolved nutrients and the sensitivity of nutrients to hydrological conditions was determined by their preferential sorption on these sediments. Second, movement of nutrients into runoff occurred more readily as soils became wetter during an event.


Synthesis of Water-Soluble Imidazolium Polyesters as Potential Nonviral Gene Delivery Vehicles.
The inherent hydrolytic reactivity of polyesters renders them excellent candidates for a variety of biomedical applications. Incorporating ionic groups further expands their potential impact, encompassing charge-dependent function such as deoxyribonucleic acid (DNA) binding, antibacterial properties, and pH-responsiveness. Catalyst-free and solvent-free polycondensation of a bromomethyl imidazolium-containing (BrMeIm) diol with neopentylglycol (NPG) and adipic acid (AA) afforded novel charged copolyesters with pendant imidazolium sites. Varying ionic content influenced thermal properties and offered a wide-range, -41 to 40 °C, of composition-dependent glass transition temperatures (Tgs). In addition to desirable melt and thermal stability, polyesters with ionic concentrations ≥15 mol % readily dispersed in water, suggesting potential as nonviral gene delivery vectors. An electrophoretic gel shift assay confirmed the novel cationic copolyesters successfully bound DNA at an N/P ratio of 4 for 50 mol % and 75 mol % charged copolyesters (P(NA50-co-ImA50) and P(NA25-co-ImA75)), and an N/P ratio of 5 for 100 mol % Im (PImA). Polyplexes exhibited insignificant cytotoxicity even at high concentrations (200 μg/mL), and a Luciferase transfection assay revealed the ionic (co)polyesters transfected DNA significantly better than the untreated controls. The successful transfection of these novel (co)polyesters inspires future imidazolium-containing polyester design.


Significance of lymph node metastasis in patients with diffuse malignant peritoneal mesothelioma.
Diffuse malignant peritoneal mesothelioma (DMPM) is a rare and invariably fatal neoplasm. Some studies have shown that cytoreductive surgery (CRS) combined with perioperative intraperitoneal chemotherapy (PIC) achieved an improved survival, as compared to historical controls. However, the significance of lymph node involvement in this disease has never been well defined. One hundred patients with DMPM underwent CRS and PIC at the Washington Hospital Center. The inclusion criteria for surgery consisted of histological diagnosis of DMPM, age <80 years and good performance status. All data were collected prospectively. Lymph node status, seven clinical variables and eight treatment-related prognostic factors were analyzed for survival. Seven patients were lymph node positive and they all died of their disease within 2 years after the surgery. The remaining 93 patients had 5- and 7-year survival of 50% and 43%, respectively. Univariate analysis showed that gender (p<0.001), peritoneal cancer index (p=0.009), lymph node status (p<0.001), extra-abdominal invasion (p=0.026), histological type (p<0.001), intraoperative blood loss (p=0.035), completeness of cytoreduction (p<0.001), intraperitoneal chemotherapy regimen (p=0.041), and redo cytoreductive surgery (p=0.022) were significant for survival. Multivariate analysis demonstrated that female gender, lymph node metastasis not detected, epithelial type, and adequate cytoreduction were independently associated with an improved survival. CRS and PIC showed an improved survival for DMPM, as compared to historical controls. Lymph node status along with three other prognostic variables was significant in the multivariate analysis.


Importance of the (nabla) D term in frequency-resolved optical diffusion imaging.
The effects of the approximation DD=0 that is often used in frequency-resolved optical diffusion imaging are examined. It is shown that this approximation can affect the performance of integral-equation-based approaches to optical diffusion imaging, such as the Born iterative method and the distorted Born iterative method. The approximation introduces errors into the calculation of data used in simulations, which can lead to misleading evaluations of reconstruction algorithms. Numerical calculations show the magnitude of these effects and the appearance of artifacts in reconstructed images when conventional inversion algorithms are applied to more accurately calculated data.


Methylome analysis and epigenetic changes associated with menarcheal age.
Reproductive factors have been linked to both breast cancer and DNA methylation, suggesting methylation as an important mechanism by which reproductive factors impact on disease risk. However, few studies have investigated the link between reproductive factors and DNA methylation in humans. Genome-wide methylation in peripheral blood lymphocytes of 376 healthy women from the prospective EPIC study was investigated using LUminometric Methylation Assay (LUMA). Also, methylation of 458877 CpG sites was additionally investigated in an independent group of 332 participants of the EPIC-Italy sub-cohort, using the Infinium HumanMethylation 450 BeadChip. Multivariate logistic regression and linear models were used to investigate the association between reproductive risk factors and genome wide and CpG-specific DNA methylation, respectively. Menarcheal age was inversely associated with global DNA methylation as measured with LUMA. For each yearly increase in age at menarche, the risk of having genome wide methylation below median level was increased by 32% (OR:1.32, 95%CI:1.14-1.53). When age at menarche was treated as a categorical variable, there was an inverse dose-response relationship with LUMA methylation levels (OR(12-14 vs. ≤11 yrs):1.78, 95%CI:1.01-3.17 and OR(≥15 vs. ≤11 yrs):4.59, 95%CI:2.04-10.33; P for trend<0.0001). However, average levels of global methylation as measured by the Illumina technology were not significantly associated with menarcheal age. In locus by locus comparative analyses, only one CpG site had significantly different methylation depending on the menarcheal age category examined, but this finding was not replicated by pyrosequencing in an independent data set. This study suggests a link between age at menarche and genome wide DNA methylation, and the difference in results between the two arrays suggests that repetitive element methylation has a role in the association. Epigenetic changes may be modulated by menarcheal age, or the association may be a mirror of other important changes in early life that have a detectable effect on both methylation levels and menarcheal age.


Increased spasticity from a fracture in the baclofen catheter caused by Charcot spine: case report.
In patients with Charcot spine, a loss of normal feedback response from the insensate spine results in spinal neuropathy. Increasing deformity, which can manifest as sitting imbalance, crepitus, or increased back pain, can result. We present the case of a patient with a high-thoracic spinal cord injury (SCI) who subsequently developed a Charcot joint at the T10-11 level that resulted in a dramatic increase in previously controlled spasticity after fracture of an existing baclofen catheter. The 68-year-old man with T4 paraplegia presented with increasing baclofen requirements and radiographic evidence of fracture of the intrathecal baclofen catheter with an associated Charcot joint with extensive bony destruction. The neuropathic spinal arthropathy caused mechanical baclofen catheter malfunction and resulting increased spasticity. The patient was found to have transected both his spinal cord and the baclofen catheter. Treatment consisted of removal of the catheter and stabilization with long-segment instrumentation and fusion from T6 to L2. Follow-up radiographs obtained a year and a half after surgery showed no evidence of hardware failure or significant malalignment. The patient has experienced resolution of symptoms and does not require oral or intrathecal baclofen. This is the only reported case of a Charcot spine causing intrathecal catheter fracture, leading to increased spasticity. This noteworthy case suggests that late spinal instability should be considered in the setting of SCI and increased spasticity.


A monoclonal antibody against antigen-specific helper factor augments T-cell help.
Antigen-specific molecules, commonly termed 'factors', have been shown to be released from helper and suppressor T cells. These factors mimic the activity of the cells that secrete them and there is much speculation about the relationship of antigen-specific factors to T-cell receptors for antigen. We have raised a variety of antisera in rabbits which were shown to react against conserved 'constant' determinants on either helper or suppressor factors independently of antigenic specificity or mouse strain of origin of the factor. In contrast, syngeneic mouse antisera were found to react with 'variable' factor determinants in an antigen-specific and mouse strain-dependent manner. These antisera thus define two regions on factor molecules, one 'variable' (related to antigen specificity) and the other 'constant' (related to function). However, potential contaminants in these antisera have limited their usefulness. Thus, we are now generating monoclonal antibodies against T-cell factors and report here the properties of a monoclonal antibody (AF3.44.4) which reacts with antigen-specific helper factors. This antibody also binds to helper T cells and, in the presence of antigen, augments helper cell induction in vitro, which, in turn, leads to enhanced antibody production in vitro. These characteristics suggest that AF3.44.4 recognizes a determinant shared by helper factor and the antigen receptor on helper T cells.


Design, synthesis, and biological evaluation of indenoisoquinoline topoisomerase I inhibitors featuring polyamine side chains on the lactam nitrogen.
The indenoisoquinolines are a class of noncamptothecin topoisomerase I inhibitors that display significant cytotoxicity in human cancer cell cultures. They offer a number of potential advantages over the camptothecins, including greater chemical stability, formation of more persistent cleavage complexes, and induction of a unique pattern of DNA cleavage sites. Molecular modeling has suggested that substituents on the indenoisoquinoline lactam nitrogen would protrude out of the DNA duplex in the ternary cleavage complex through the major groove. This indicates that relatively large substituents in that location would be tolerated without compromising biological activity. As a strategy for increasing the potencies and potential therapeutic usefulness of the indenoisoquinolines, a series of compounds was synthesized containing polyamine side chains on the lactam nitrogen. The rationale for the synthesis of these compounds was that the positively charged ammonium cations would increase DNA affinity through electrostatic binding to the negatively charged DNA backbone, and the polyamines might also facilitate cellular uptake by utilization of polyamine transporters. The key step in the synthesis involved the condensation of Schiff bases, containing protected amine side chains, with substituted homophthalic anhydrides, to afford cis-3-aryl-4-carboxy-1-isoquinolones. These isoquinolones were then converted to indenoisoquinolines with thionyl chloride. Although monoamines were much more potent than the lead compound, no significant increase in potency was observed through incorporation of additional amino groups in the side chain. However, one of the monoamine analogues, which features a bis(2-hydroxyethyl)amino group in the side chain, proved to be one of the most cytotoxic indenoisoquinoline synthesized to date, with a GI50 mean-graph midpoint (MGM) of 0.07 microM in the NIH human cancer cell culture screen, and topoisomerase I inhibitory activity comparable to that of camptothecin.


Maternal use of oral contraceptives and risk of fetal death.
Intrauterine exposure to artificial sex hormones such as oral contraceptives may be associated with an increased risk of fetal death. Between 1996 and 2002, a total of 92 719 women were recruited to The Danish National Birth Cohort and interviewed about exposures during pregnancy. Outcome of pregnancy was identified through linkage to the Civil Registration System and the National Discharge Registry. The authors analysed the risk of fetal death after recruitment to the cohort by using proportional hazards regression models with gestational age as the underlying time scale. In total, 1102 (1.2%) women took oral contraceptives during pregnancy. Use of combined oestrogen and progesterone oral contraceptives (COC) or progesterone-only oral contraceptives (POC) during pregnancy was not associated with increased hazard ratios of fetal death compared with non-users, HR 1.01 [95% CI 0.71, 1.45] and HR 1.37 [95% CI 0.65, 2.89] respectively. Neither use of COC nor POC prior to pregnancy was associated with fetal death. Stratification by maternal age and smoking showed elevated risks of fetal death for women <30 years and smokers using oral contraception during pregnancy, but the interactions were not significant. In conclusion, there was no evidence that oral contraceptive use before or during pregnancy is associated with an increased risk of fetal death.


Two-year outcomes after transcatheter or surgical aortic-valve replacement.
The Placement of Aortic Transcatheter Valves (PARTNER) trial showed that among high-risk patients with aortic stenosis, the 1-year survival rates are similar with transcatheter aortic-valve replacement (TAVR) and surgical replacement. However, longer-term follow-up is necessary to determine whether TAVR has prolonged benefits. At 25 centers, we randomly assigned 699 high-risk patients with severe aortic stenosis to undergo either surgical aortic-valve replacement or TAVR. All patients were followed for at least 2 years, with assessment of clinical outcomes and echocardiographic evaluation. The rates of death from any cause were similar in the TAVR and surgery groups (hazard ratio with TAVR, 0.90; 95% confidence interval [CI], 0.71 to 1.15; P=0.41) and at 2 years (Kaplan-Meier analysis) were 33.9% in the TAVR group and 35.0% in the surgery group (P=0.78). The frequency of all strokes during follow-up did not differ significantly between the two groups (hazard ratio, 1.22; 95% CI, 0.67 to 2.23; P=0.52). At 30 days, strokes were more frequent with TAVR than with surgical replacement (4.6% vs. 2.4%, P=0.12); subsequently, there were 8 additional strokes in the TAVR group and 12 in the surgery group. Improvement in valve areas was similar with TAVR and surgical replacement and was maintained for 2 years. Paravalvular regurgitation was more frequent after TAVR (P<0.001), and even mild paravalvular regurgitation was associated with increased late mortality (P<0.001). A 2-year follow-up of patients in the PARTNER trial supports TAVR as an alternative to surgery in high-risk patients. The two treatments were similar with respect to mortality, reduction in symptoms, and improved valve hemodynamics, but paravalvular regurgitation was more frequent after TAVR and was associated with increased late mortality. (Funded by Edwards Lifesciences; ClinicalTrials.gov number, NCT00530894.).